Expression Vectors

Expression Vectors

APOBEC-Related >> All

DATA SHEET 

Reagent: 

pcDNA3.1-AID-V5-6XHis 

Catalog Number: 10099 

Lot Number: 4 040905 

Release Category: A 

Provided: 2.0 µg plasmid DNA in 2.0 µl of TE buffer. The plasmid was amplified in HB 101 

competent cells (Invitrogen Corp). 

Description: 

The cloning insert was obtained from Open Biosystems (Huntsville, AL). Insert for human 

Activation-induced (cytidine) deaminase (AID) was cloned into mammalian expression 

vector pcDNA3 with a V5 epitope tag from SV5 paramyxovirus and a polyhistidine 

epitope at the C terminus. Plasmid DNA was transfected into 293T cells and expression of 

human AID was detected with a monoclonal anti-V5 antibody. Expression of human AID 

is driven by the CMV promoter. 

Cloning Site: 

TOPO Cloning Technology was used following manufacturer’s instructions (Invitrogen 

Corp). The size of the insert is 597 bp. 

Cloning Vector: 

The cloning vector is pcDNA3.1D/V5-His-Topo. The size of the cloning vector including 

the insert is 6111 bp. 

Recommended 

Storage: 

4°C 

Contributor: Drs. B. Matija Peterlin and Yong-Hui Zheng 

References: Yong-Hui Zheng, Dan Irwin, Takeshi Kurosu, Kenzo Tokunaga, Tetsutaro Sata and B. 

Matija Peterlin. Human APOBEC3F Is Another Host Factor That Blocks Human 

Immunodeficiency Virus Type 1 Replication. J. Virology 78:6073-6076, 2004. 

ALL RECIPIENTS OF THIS MATERIAL MUST COMPLY WITH ALL APPLICABLE BIOLOGICAL, CHEMICAL, 

AND/OR RADIOCHEMICAL SAFETY STANDARDS INCLUDING SPECIAL PRACTICES, EQUIPMENT, 

FACILITIES, AND REGULATIONS. NOT FOR USE IN HUMANS. 

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NOTE: 

Acknowledgment for publications should read "The following reagent was obtained 

through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: 

pcDNA3.1-AID-V5-6XHis from Drs. B. Matija Peterlin and Yong-Hui Zheng." Also include 

the reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1-APOBEC3F-V5-6XHis 



Release 

Category: 

A 

Provided: 

2.0 µg plasmid DNA in 2.0 µl TE buffer. The plasmid was amplified in HB 101 competent 

cells (Invitrogen Corp). 

Description: 


APOBEC3F was cloned into mammalian expression vector pcDNA3 with a V5 epitope tag 

from SV5 paramyxovirus and a polyhistidine epitope at the C terminus. Plasmid DNA was 

transfected into 293T cells and expression of human APOBEC3F was detected with with a 

monoclonal anti-V5 antibody. Expression of human APOBEC3F is driven by the CMV 

promoter. 

Cloning Site: 

TOPO Cloning Technology was used following manufacture’s instruction (Invitrogen Corp). 

The size of the insert is 1122 bp. 




Recommended 

Storage: 

4°C 








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NOTE: 



pcDNA3.1-APOBEC3F-V5-6XHis from Drs. B. Matija Peterlin and Yong-Hui Zheng." Also 

include the reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1-APOBEC3C-V5-6XHis 



Release 

Category: 

A 

Provided: 


cells (Invitrogen Corp.). 

Description: 


APOBEC3C was cloned into mammalian expression vector pcDNA3 with a V5 epitope tag 


transfected into 293T cells and expression of human APOBEC3C was detected with with a 

monoclonal anti-V5 antibody. Expression of human APOBEC3C is driven by the CMV 

promoter. 

Cloning Site: 






Recommended 

Storage: 

4°C 








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NOTE: 



pcDNA3.1-APOBEC3C-V5-6XHis from Drs. B. Matija Peterlin and Yong-Hui Zheng." Also 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1-APOBEC1-V5-6XHis 



Release 

Category: 

A 

Provided: 


cells (Invitrogen Corp), 

Description: 

The cloning insert was obtained from Dr. N. O. Davidson (Washington University, St 

Louis, MO). Insert for human APOBEC1 was cloned into mammalian expression vector 

pcDNA3 with a V5 epitope tag from SV5 paramyxovirus and a polyhistidine epitope at 

the C terminus. Plasmid DNA was transfected into 293T cells and expression of human 

APOBEC1 was detected with a monoclonal anti-V5 antibody. Expression of human 

APOBEC1 is driven by the CMV promoter. 

Cloning Site: 

TOPO Cloning Technology was used following Manufacture’s instructions (Invitrogen 

Corp.) The size of the insert is 711 bp. 




Recommended 

Storage: 

4°C 








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NOTE: 



pcDNA3.1-APOBEC1-V5-6XHis from Drs. B. Matija Peterlin and Yong-Hui Zheng." Also 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1-APOBEC2-V5-6XHis 



Release 

Category: 

A 

Provided: 



Description: 


APOBEC2 was cloned into mammalian expression vector pcDNA3 with a V5 epitope tag 


transfected into 293T cells and expression of human APOBEC2 was detected with with a 

monoclonal anti-V5 antibody. Expression of human APOBEC2 is driven by the CMV 

promoter. 

Cloning Site: 






Recommended 

Storage: 

4°C 








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NOTE: 



pcDNA3.1-APOBEC2-V5-6XHis from Drs. B. Matija Peterlin and Yong-Hui Zheng." Also 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pc-Mu-APOBEC3G-HA 



Release 

Category: 

C 

Provided: 2 µg plasmid DNA in 50 µl TE 

Description: 

RNA was prepared from C57BL/6 mouse spleen. cDNA was synthesized with Superscript 

II reverse transcriptase (Invitrogen) primed by oligo dT (Invitrogen). The plasmid 

expresses Mu-APOBEC3G under the control of a CMV promoter and contains a C-terminal 

HA epitope tag (see Mariani R, et al. for details). Note that the plasmid can be 

transformed in HB101 cells and better plasmid yield could be obtained if the bacteria is 

grown using the chloramphenicol amplification step. Resistance marker is ampicillin. 

Click here to see the sequence file 

Cloning Site: EcoRI and XhoI cloning site. The size of the insert is 1320 bp. 


The cloning vector is pcDNA3.1(+), the size of the cloning vector including the insert is 

6700 bp. 

Special 

Characteristics: 

Cloning Strategy: The cloning vector is pcDNA3.1(+). 

Recommended 

Storage: 

4°C 

Contributor: Dr. Nathaniel Landau. 




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References: Mariani R, Chen D, Schrofelbauer B, Navarro F, Konig R, Bollman B, Munk C, 

Nymark-McMahon H, Landau NR. Species-specific exclusion of APOBEC3G from HIV-1 

virions by Vif. Cell 114:21-31, 2003. 

NOTE: 



pc-Mu-APOBEC3G-HA from Dr. Nathaniel Landau." Also include the reference cited above 

in any publications. Scientists at for-profit institutions or who intend commercial 

use of this reagent must contact Pam Bock, Office of Technology Management, 

Salk Institute, La Jolla, CA 92037, Tel: 858-453-4100, ext. 2006, Email: 

Bock@salk.edu before the reagent can be released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1-mAPOBEC3G-V5-6XHis (murine APOBEC3G) 


Lot Number: 040904 

Release 

Category: 

A 

Provided: 



Description: 

The cloning insert was obtained from Open Biosystems (Huntsville, AL). Insert for murine 

APOBEC3G was cloned into mammalian expression vector pcDNA3 with a V5 epitope tag 


transfected into 293T cells and expression of murine APOBEC3G was detected with with a 

monoclonal anti-V5 antibody. Expression of murine APOBEC3G is driven by the CMV 

promoter. 

Cloning Site: 

TOPO Cloning Technology was used following manufacturer’s instruction (Invitrogen 

Corp). The size of the insert is 1290 bp. 




Recommended 

Storage: 

4°C 








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NOTE: 



pcDNA3.1-mAPOBEC3G-V5-6XHis (murine APOBEC3G) from Drs. B. Matija Peterlin and 

Yong-Hui Zheng." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1-APOBEC3G-V5-6XHis 



Release 

Category: 

A 

Provided: 



Description: 


APOBEC3G was cloned into mammalian expression vector pcDNA3 with a V5 epitope tag 


transfected into 293T cells and expression of human APOBEC3G was detected with with a 

monoclonal anti-V5 antibody. Expression of human APOBEC3G is driven by the CMV 

promoter. 

Genebank Acc# BC024268 

Cloning Site: 






Recommended 

Storage: 

4°C 








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NOTE: 



pcDNA3.1-APOBEC3G-V5-6XHis from Drs. B. Matija Peterlin and Yong-Hui Zheng." Also 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA-APO3G 



Release Category: C 

Provided: 1ml glycerol stock of transformed ampicillin resistant DH5ß competent cells. 

Description: 

This plasmid expresses human APOBEC3G under the control of the CMV promoter. 

Expresses a 409 amino acid protein (384 residues APOBEC3G plus 25 residues MycHis 

epitope tag). 

Note: this clone contains two nucleotide changes (G1448→A) & (G2071→T) in APOBEC3G 

resulting in two amino acid changes (S162N) & (D370Y) relative to the GenBank entries for 

APOBEC3G (NM_021822) and MDS019 (AF182420). 

Cloning Site: 

Cloned into the EcoRI/HindIII sites of pcDNA3.1(-)/MycHis A (Invitrogen Corp., 

Carlsbad CA). The size of the insert is 1162 bp. 


The cloning vector is pcDNA3.1(-)/MycHis A; the size of the cloning vector including 


Special 


Expresses biologically active human APOBEC3G. 

Recommended 

Storage: 

-70°C 

Contributor: Drs. Klaus Strebel and Sandra Kao. 




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References: 

Kao S, Khan MA, Miyagi E, Plishka R, Buckler-White A, Strebel K. The human 

immunodeficiency virus type 1 Vif protein reduces intracellular expression and inhibits 

packaging of APOBEC3G (CEM15), a cellular inhibitor of virus infectivity. J Virol 

77:11398-11407, 2003. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcDNA-APO3G 

from Drs. Klaus Strebel and Sandra Kao." Also include the reference cited above in 

any publications. 

Scientists at for-profit institutions or who intend commercial use of this 

reagent must contact Dr. Sally Hu at the NIH Office of Technology Transfer, 

Email: hus@mail.nih.gov, Phone: 301-435-5606, before the reagent can be 

released. Please specify the name and a description of the intended use of the 

reagent. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pHIV-APO3G 



Release 

Category: 

C 

Provided: 1.0 µg plasmid DNA in water (0.1 µg/ུl) 

Description: 

This plasmid expresses human APOBEC3G under the control of the HIV-1 promoter. Requires Tat for 

expression. APOBEC3G was cloned by PCR from pcDNA-APO3G (Cat# 9904). Expresses a 409 

amino acid protein (384 residues APOBEC3G plus 25 residues MycHis epitope tag). Note: this clone 

contains two nucleotide changes (G1448→A) & (G2071→T) in APOBEC3G resulting in two amino acid 

changes (S162N) & (D370Y) relative to the GenBank entries for APOBEC3G (NM_021822) and 

MDS019 (AF182420). Also, note that this plasmid has an extra third band when digested but this does 

not affect the expression of the APOBEC3G protein in a Tat dependent manner. 

Cloning Site: 

APOBEC3G gene was cloned into the BssHII/XhoI sites of pNL4-3 (Cat# 114). The size of 


Cloning Vector: The cloning vector is pNL4-3; the size of the cloning vector including the insert is 7941 bp. 

Special 


Expresses biologically active human APOBEC3G under the control of the HIV-1 LTR. 

Protein expression is Tat-dependent. Vector restricts APOBEC3G expression to 

HIV-expressing cells. 

Recommended 

Storage: 

-70°C 

Contributor: Drs. Klaus Strebel and Sandra Kao. 




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References: 

Kao S, Khan MA, Miyagi E, Plishka R, Buckler-White A, Strebel K. The human 

immunodeficiency virus type 1 Vif protein reduces intracellular expression and inhibits 

packaging of APOBEC3G (CEM15), a cellular inhibitor of virus infectivity. J Virol 

77:11398-11407, 2003. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pHIV-APO3G from 

Drs. Klaus Strebel and Sandra Kao." Also include the reference cited above in any 

publications. 

Scientists at for-profit institutions or who intend commercial use of this reagent 

must contact Dr. Sally Hu at the NIH Office of Technology Transfer, 



reagent. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1-APOBEC3G-HA 



Release Category: E 

Provided: 6 µg of purified DNA at 1.2 µg/µL in TE buffer. 

Description: 

pcDNA3.1-APOBEC3G-HA expresses APOBEC3G/CEM15 with a C-terminal triple HA tag 

in mammalian cells. 

Cloning Site: 

The insert was excised from expression vector pNG/C15 (Malim Lab), by cutting with 

XhoI (5’) and EcoRI (3’). It was inserted into the corresponding sites of pcDNA 3.1 (-). 

The size of the APOBECG3-HA insert is ~1.7 kb. 


The cloning vector is pcDNA 3.1 (-) (Invitrogen). The size of the cloning vector 

including the insert is ~ 7.1 kb. 

Special 


The insert was derived from the retroviral expression vector, pNG/C15 from the 

laboratory of Michael Malim. Expression of APOBEC3G-HA is constitutive and under 

control of the CMV immediate early promoter. 

This expression vector for APOBEC3G is useful for transient expression in HEK 293 and 

HEK 293T cells. The C-terminal triple HA tag works well in immunoblotting and 

immunoprecipitation. 

Plasmid map and sequence file lot 042043. 

Recommended 

Storage: 

Keep at -20°C or lower. Avoid freeze-thaw cycles as reagent degradation may result. 

Contributor: Dr. Warner C. Greene. 




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References: 

Stopak K, de Noronha C, Yonemoto W, Greene WC. HIV-1 Vif blocks the antiviral 

activity of APOBEC3G by impairing both its translation and intracellular stability. Mol 

Cell 12:591-601, 2003. Sheehy AM, Gaddis NC, Choi JD, Malim MH. Isolation of a 

human gene that inhibits HIV-1 infection and is suppressed by the viral Vif protein. 

Nature 8:646-650, 2002. 

NOTE: 



pcDNA3.1-APOBEC3G-HA from Dr. Warner C. Greene." Also include the reference cited 

above in any publications. 

Recipient must not use or incorporate the reagent for commercial purposes. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

phAPOBEC3B-HA 


Lot Number: hApo3B+3xHA 


Provided: 5 µg plasmid DNA/vial (5µg/5µl) 

Description: It contains a 1275 bp kpnI/xhoI insert encoding the hAPOBEC3B gene linked to 3 

HA-tags. Sequence information is available upon request. 

pcDNA3 vector 

Cloning Site: The cloning site of the APOBEC3B cDNA is kpnI/xhoI and the size of the insert is 1275 

bp. HindIII site is immediately 5' to the KpnI site (5'- GGTACC-3') in the cloning vector 

and can also be used to excise this cDNA. 

Cloning Vector: The cloning vector is pcDNA3 and the size of the cloning vector including the insert is 6.6 

Kb. 

Special 


It is a cytidine deaminase which inhibits HIV-1 and is not Vif sensitive. 

Recommended 

Storage: 

4-8°C 

Contributor: Dr. Bryan R. Cullen 




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References: 

Doehle B.P., Schäfer A., Wiegand H.L., Bogerd H.P. and Cullen B.R. Differential 

sensitivity of murine leukemia virus to APOBEC3-mediated inhibition is governed by virion 

exclusion. J. Virology 79: 8201-8207, 2005. 

Doehle B.P., Schäfer A. and B.R. Cullen. Human APOBEC3B is a potent inhibitor of HIV-1 

infectivity and is resistant to HIV-1 Vif. Virology 339: 281-8, 2005. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: phAPOBEC3B-HA 

from Dr. Bryan R. Cullen." Also include the reference cited above in any publications. 


must contact the donor, Dr. Bryan R. Cullen, at the department of Molecular 

Genetics and Microbiology, Duke University Center for Virology, Duke University 

Medical Center, Box 3025, Room 426 CARL Building, Research Drive, Durham, 

NC 27710, Tel.: 919-684-3369, Fax: 919-681-8979, E-mail: 

culle002@mc.duke.edu, before the reagent can be released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1 Human APOBEC3G-Myc-6XHis 




Provided: 1 ml transformed DH5α containing 0.8 ml of bacterial culture plus 0.2 ml glycerol. 

Description: 

The coding sequence of human APOBEC3G was cloned by RT-PCR using total RNA 

from H9 cells. The 1152 bp fragment was cloned into the vector with the Myc and 

6XHis epitopes encoded at the C-terminus of APOBEC3G. The expression is driven by 

the CMV promoter. 

Cloning Site: Xho I–5' and Sfu I–3' cloning sites. The size of the insert is 1152 bp. 


The cloning vector is pcDNA3.1/Myc-HisC (Invitrogen). The size of the cloning vector 

including the insert is 6652 bp. 

Special 


Cloning Strategy: The cloning vector is pcDNA3.1/Myc-HisC (Invitrogen). 

Human APOBEC3G Sequence 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. David Kabat. 

References: 

Marin M, Rose KM, Kozak SL, Kabat D. HIV-1 Vif protein binds the editing enzyme 

APOBEC3G and induces its degradation. Nat Med 9:1398-1403, 2003. 




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NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcDNA3.1 

Human APOBEC3G-Myc-6Xhis from Dr. David Kabat." Also include the reference cited 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pCMV4-HA-APOBEC3G 




Provided: 5.0 μg plasmid DNA in 1 x TE (5.0 μl). 

Description: 

pCMV4-HA-APOBEC3G was created to express APOBEC3G/CEM15 with an N-terminal 

triple HA tag in mammalian cells. The insert was amplified; using PCR from a cDNA 

library from HIV-1 infected H9 cells. Expression of HA-APOBEC3G is constitutive and 

under control of the CMV immediate early promoter. 

Cloning Site: 

APOBEC3G cDNA was amplified by PCR from an H9 cDNA library and then inserted into 

the HindIII (5’) and XbaI (3’) sites of the pCMV4 vector. The size of the APOBECG3-HA 

insert is ~1.2 kb. 


The cloning vector is pCMV4 (Andersson S, J Biol Chem 264:8222-8229, 1989). The 

size of the cloning vector including the insert is ~ 6.1 kb. 

Special 


Host: DH5α or similar host. 

This expression vector for APOBEC3G is useful for transient expression in HEK 293 and 

HEK 293T cells. The N-terminal triple HA tag is useful for immunoblotting and 

immunoprecipitation. 

Recommended 

Storage: 

-20°C 

Contributor: Dr. Warner C. Greene. 




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References: 

Stopak K, de Noronha C, Yonemoto W, Greene WC. HIV-1 Vif blocks the antiviral 

activity of APOBEC3G by impairing both its translation and intracellular stability. Mol 

Cell 12:591 601, 2003. 

NOTE: 



pCMV4-HA-APOBEC3G from Dr. Warner C. Greene.:" Also include the reference cited 






REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1-APOBEC3DE-V5-6xHIS 




Provided: 5 μl purified plasmid DNA (1.2 μg/μl). 

Description: 


AID was cloned into mammalian expression vector pcDNA3 with a V5 epitope tag from 

SV5 paramyxovirus and a polyhistidine epitope at the C terminus. Plasmid DNA was 

transfected into 293T cells and expression of human APOBEC3DE was detected with a 

monoclonal anti-V5 antibody. Expression of human APOBEC3DE is driven by the CMV 

promoter. 

Cloning Site: TA cloned 


pcDNA3.1D/V5-His-Topo vector (Topo cloning system). Size of insert is 1158bp. Size of 

vector and insert is 6672bp. 

Special 


Host Strain: Grown in HB101 

Recommended 

Storage: 

-20°C 

Contributor: Dr. Yong-Hui Zheng 

References: 

Ying Dang, Xiaojun Wang, Walter J. Esselman, and Yong-Hui Zheng. Identification of 

APOBEC3DE as Another Antiretroviral Factor from the Human APOBEC Family. J. Virol. 

2006 80: 10522-10533. [Abstract] [Full Text] [PDF] 




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NOTE: 

Acknowledgment for publications should read “The following reagent was obtained 


pcDNA3.1-APOBEC3DE-V5-6xHIS (Cat#11433) from Dr. Yong-Hui Zheng.” Also include 


Scientists at for-profit institutions or who intend commercial use of Release 

Category C Reagents (Cat# 11433) must contact Dr. Yong-Hui Zheng, 

Department of Microbiology and Molecular Genetics, 4174 Biomedical Physical 

Sciences, Tel: 517 355 6463 Ext.1528, Email: zhengyo@msu.edu, before the 

reagent can be released. 




REV: 11/08/2015 Page 2 of 2

BIV >> ENV

DATA SHEET 

Reagent: 

BIV-env8 



Release Category: B 

Provided: 1 vial transformed RRI bacteria. 

Description: 

The 0.4 kb TaqI fragment (nt 7076-7478) was cloned into the blunt-ended BamHI site 

of pATH-1. 

Special 


This clone expresses the BIV env major antigenic domain as a fusion protein. The 

expressed protein is recognized by BIV-infected animal serum. 

Contributor: Dr. Charles Wood, University of Nebraska, Lincoln. 

References: 

Atkinson B, Liu ZQ, Wood C. Use of bacterial trpE fusion vectors to express and 

characterize the bovine immunodeficiency-like virus core protein. J Virol Meth 

36:35-49, 1992. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: BIV-env8 from 

Dr. Charles Wood, University of Nebraska, Lincoln.." Also include the reference cited 

above in any publications. This reagent is not to be used for commercial purposes. 




REV: 11/08/2015 Page 1 of 1

BIV >> GAG

DATA SHEET 

Reagent: 

BIV-gag3 





Description: 

The StuI-BamHI fragment derived from BIV gag was blunt-ended, then ligated into 

pATH3. 

Special 


This clone expresses the major BIV gag antigenic determinant as a fusion protein. The 

expressed protein has been used as an antigen in Western blots to screen for 

antibodies against BIV in infected animals. 


References: 



36:35-49, 1992. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: BIV-gag3 from 






REV: 11/08/2015 Page 1 of 1

DATA SHEET 

Reagent: 

BIV-gag5 





Description: 

The PstI fragment (nt 1130-1642) BIV gag was subcloned into pUC8 to generate pUC 

gag-3. The EcoRI-HindIV fragment from pUC gag-3 was then ligated into pATH3 to 

generate BIV-gag5. 

Special 


This clone expresses the major BIV gag antigenic determinant as a fusion protein. The 

expressed protein has been used as an antigen in Western blots to screen for 

antibodies against BIV in infected animals. 


References: 



36:35-49, 1992. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: BIV-gag3 from 






REV: 11/08/2015 Page 1 of 1

CD4 or CD8 >> All

DATA SHEET 

Reagent: 

T4-pMV7 




Provided: 5 μg purified DNA, 1 μg/μL in TE. 

Description: 

A 3 kb pT4B EcoRI-BamHI site cDNA insert was cloned into the EcoRI site of pMV7. pT4B 

(Catalog #157) encodes the human CD4 receptor and was originally cloned from human 

peripheral T lymphocytes (GenBank M12807.1). pMV7 contains two LTR repeats of 

Moloney murine sarcoma virus spanning a unique EcoRI site. Also contains the bacterial 

neomycin phosphotransferase gene (neo) fused to the HSV tk promoter, both located 

downstream of the cloning site. 

T4-pMV7 is a recombinant retroviral expression vector expressing the human CD4 

receptor in mammalian cells. pMV7 contains two LTR repeats of Moloney murine sarcoma 

virus spanning a unique EcoRI cloning site. pMV7 also contains the bacterial neomycin 

phosphotransferase gene (neo) fused to the HSV thymidine kinase promoter (tk), both 

located downstream of the cloning site. 

Plasmid Map 

Special 


Transfection of T4-pMV7 to a retrovirus helper cell line (AM or psi 2) results in the 

production of replication-defective recombinant retrovirus. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Richard Axel. 




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References: 

Maddon PJ, Dalgleish AG, McDougal JS, Clapham PR, Weiss RA, Axel R. The T4 gene 

encodes the AIDS virus receptor and is expressed in the immune system and the brain. 

Cell 47:333-348, 1986. Kirschmeier PT, Housey GM, Johnson MD, Perkins AS, Weinstein 

IB. Construction and characterization of a retroviral vector demonstrating efficient 

expression of cloned cDNA sequences. DNA 7:219-225, 1988. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: T4-pMV7 from 

Dr. Richard Axel." Also include the appropriate references cited above in any 

publications. The CD4 gene is in pT4B (catalog #157). 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

T8-pMV7 




Provided: 1 mL glycerol stock (HB101). Ampicilin resistant. 

Description: 

A 1.5 kb cDNA insert from pT8F1 (Catalog #179) was cloned into the EcoRI site of 

pMV7. pT8F1 encodes the human CD8 receptor and was originally cloned from the 

human T-cell leukemia Fro 2.2. T8-pMV7 contains two LTR repeats of Moloney murine 

sarcoma virus spanning a unique EcoRI site. Also contains the bacterial neomycin 

phosphotransferase gene (neo) fused to the HSV tk promoter, both located downstream 

of the cloning site. 

T8-pMV7 is a recombinant retroviral expression vector expressing the human CD8 

receptor in mammalian cells. pMV7 contains two LTR repeats of Moloney murine sarcoma 

virus spanning a unique EcoRI cloning site. pMV7 also contains the bacterial neomycin 

phosphotransferase gene (neo) fused to the HSV thymidine kinase promoter (tk), both 

located downstream of the cloning site. 

Plasmid Map 

Special 


T8-pMV7 contains full length cDNA insert encoding CD8. Transfection of T8-pMV7 to a 

retrovirus helper cell line (AM or psi 2) results in the production of replication-defective 

recombinant retrovirus. The CD8 gene is also in pT8F1 (catalog #179). 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Richard Axel. 




REV: 11/08/2015 Page 1 of 2

References: 

Maddon PJ, Dalgleish AG, McDouga, JS, Clapham PR, Weiss RA, Axel R. The T4 gene 

encodes the AIDS virus receptor and is expressed in the immune system and the brain. 

Cell 47:333-348, 1986. 

Kirschmeier PT, Housey GM, Johnson MD, Perkins AS, Weinstein IB. Construction and 

characterization of a retroviral vector demonstrating efficient expression of cloned cDNA 

sequences. DNA 7:219-225, 1988. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: T8-pMV7 from 

Dr. Richard Axel." Also include the references cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pc.Rh-CD4 



Release Category: D 

Provided: 1 mL transformed TOP-10 bacteria (glycerol stock). 

Description: This expression vector produces rhesus CD4 

Cloning Site: 

The insert is cloned into the CMV promoter driven expression vector pcDNAI/Amp as a 

BamHI-XhoI 1.4 kb fragment. 

Cloning Vector: pcDNAI/Amp, 4.8 kb. 

Special 


Expressed CD4 can be detected by FACS analysis. 

Other available clones in this set: pcRh-CXCR4 (Cat #3601), pcRh-CCR5 (Cat #3602) 

The GenBank accession number for Rh-CCR5 is U73739; the GenBank accession 

number for Rh-CXCR4 is U73740. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Preston Marx. 

References: 

Chen Z, Zhou P, Ho DD, Landau NR, Marx PA. Genetically divergent strains of simian 

immunodeficiency virus use CCR5 as a coreceptor for entry. J Virol 71:2705–2714, 

1997. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: (specify clone) 

from Dr. Preston Marx." Also include the reference cited above in any publications. 

Research Chart: 

Clone 

Cat. 

No. 

Lot No. 

pc.Rh-CD4 3600 4 99040 

pc.Rh-CXCR4 3601 

3 97100 4 

99041 

pc.Rh-CCR5 3602 4 99042 

Description 

Rhesus CD4 is cloned into pcDNA-I amp as a 

BamHI-XhoI fragment. 

Rhesus CXCR4 is cloned into pcDNA-I amp as 

an EcoRI-XhoI fragment. 

Rhesus CCR5 is cloned into pcDNA-I amp as a 





REV: 11/08/2015 Page 2 of 2

Chemokine Receptors >> CCR2

DATA SHEET 

Reagent: 

pcCCR2B 

Catalog 

Number: 

3322 


Release 

Category: 

C 

Provided: 29 µg purified plasmid DNA in TE buffer at 5.8 mg/mL. 

Description: 

Chemokine receptor expression vector, CCR2b. The cDNA encoding CCR2b was amplified 

from activated PBMC RNA using primers hybridizing to the 5' and 3' untranslated regions of 

CCR2b. The PCR product was digested with BamHI and SalI and cloned into the BamHI 

and XhoI sites of pcDNAI/amp (SalI and XhoI are compatible ends for ligation). Constructs 

are selected for by adding Ampicillin (100 µg/mL) to the growth medium. Chloramphenicol 

(100 µg/ml) should be added 4.5–5.5 hours (OD 600 = 1.0) after seeding the culture. The 

plasmid can be transformed into E. coli strain HB101. 

CCR-2b primer sequences: 

CCR-2B.5'BamHI (35mer): 5'-GCTCAGGATCCTGAGACAAGCCACAAGCTGAACAG-3'. 

CCR-2B.3'XhoI (34mer): 5'-GTGCCTCTAGACTGAATGCGTGAGCCCTTTGCTC-3'. 

Cloning Site: BamHI and XhoI (5'-3'). The size of the insert is approximately 1.1 kb. 

Cloning Vector: pcDNAI/amp (Invitrogen). 

Special 


Plasmid Map 

Recommended 

Storage: 

-20°C 

Contributor: Dr. Nathaniel Landau, Aaron Diamond AIDS Research Center, The Rockefeller University. 




REV: 11/08/2015 Page 1 of 2

References: 

Morgenstern JP, Land H. Advanced mammalian gene transfer: high titre retroviral vectors 

with multiple drug selection markers and a complementary helper-free packaging cell line. 

Nucleic Acids Res 18:3587-3596, 1990. 

Deng H, Liu R, Ellmeier W, Choe S, Unutmaz D, Burkhart M, Di Marzio P, Marmon S, 

Sutton RE, Hill CM, Davis CB, Peiper SC, Schall TJ, Littman DR, Landau NR. Identification of 

a major co-receptor for primary isolates of HIV-1. Nature 381:661-666, 1996. 

NOTE: 

Acknowledgment for publications should read "The following reagent was obtained through 

the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcCCR2b (Cat #3322) from 

Dr. Nathaniel Landau." Also include the references cited above in any publications. 

Patent-pending. Requests from commercial organizations must be directed to the 

New York University Medical Center, ATTN: Office of Industrial Liaison, 550 First 

Avenue, New York, NY 10016, TEL: 212-263-8178. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pBABE.CCR2B 

Catalog 

Number: 

3328 


Release 

Category: 

C 

Provided: 9.5 µg purified plasmid DNA in TE buffer at 1.9 mg/mL. 

Description: 

Chemokine receptor expression vector, CCR2b. The cDNA encoding CCR2b was amplified 

from activated PBMC RNA using primers hybridizing to the 5' and 3' untranslated regions 

of CCR2b. The PCR product was digested with BamHI and SalI and cloned into 

pBABE-puro. Constructs are selected for by adding Ampicillin (100µg/mL) to the growth 

medium. Chloramphenicol (100 µg/ml) should be added 4.5–5.5 hours (OD 600 = 1.0) 

after seeding the culture. The plasmid can be transformed into E. coli strain HB101. 

CCR-2b primer sequences: 

CCR-2B.5'BamHI (35mer): 5'-GCTCAGGATCCTGAGACAAGCCACAAGCTGAACAG-3'. 

CCR-2B.3'SalI (34mer): 5'-GTGCCTCTAGACTGAATGCGTGAGCCCTTTGCTC-3'. 

Plasmid Map 

Cloning Site: BamHI and SalI (5'-3'). The size of the insert is approximately 1.1 kb. 

Cloning Vector: pBABE-puro (Morgenstern & Land, 1990). 

Recommended 

Storage: 

-20°C 





REV: 11/08/2015 Page 1 of 2

References: 





Sutton RE, Hill CM, Davis CB, Peiper SC, Schall TJ, Littman DR, Landau NR. Identification 

of a major co-receptor for primary isolates of HIV-1. Nature 381:661-666, 1996. 

NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pBABE.CCR2b (Cat #3328) 

from Dr. Nathaniel Landau." Also include the references cited above in any publications. 







REV: 11/08/2015 Page 2 of 2


DATA SHEET 

Reagent: 

pBABE.CCR3 



Release 

Category: 

C 

Provided: 100 µL (1 µg) purified plasmid DNA in TE buffer at 10 µg/mL. 

Description: 

Chemokine receptor expression vector, CCR3. The cDNA encoding CCR3 was amplified 


of CCR3. The PCR product was first digested with EcoRI and XbaI (blunted) and cloned 

into pcDNA, then excised and cloned into the EcoRI and SalI (blunted) sites of 

pBABE-puro. Constructs are selected for by adding Ampicillin (100ug/mL) to the growth 


after seeding the culture. 

CCR-3 primer sequences: 

CCR-3.5'EcoRI (34mer): 5'-GACTCGAATCCTTCTTCTATCACAGGGAGAAGTG-3'. 

CCR-3.3'XbaI (32mer): 5'-GTGCCTCTAGACTGGAAGTTTGAAGGACTGTT-3'. 

Cloning Site: EcoRI and SalI (5'-3'). The size of the insert is approximately 1.1 kb. 


Special 


Plasmid Map 

Recommended 

Storage: 

-20°C 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pBABE.CCR3 (Cat# 3329) 








REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcCCR3 



Release 

Category: 

C 


Description: 



of CCR3. The insert was digested with EcoRI and XbaI and cloned into pcDNAI/amp 

(Invitrogen). Constructs are selected for by adding Ampicillin (100 µg/mL) to the growth 


after seeding the culture. The plasmid can be transformed in E. coli strain HB101. 


CCR-3.5'EcoRI (34mer): 5'-GACTCGAATCCTTCTTCTATCACAGGGAGAAGTG-3'. 

CCR-3.3'XbaI (32mer): 5'-GTGCCTCTAGACTGGAAGTTTGAAGGACTGTT-3'. 

Cloning Site: EcoRI and XbaI (5'-3'). The size of the insert is approximately 1.1 kb. 


Special 


Plasmid Map 

Recommended 

Storage: 

-20°C 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcCCR3 (Cat #3323) from 








REV: 11/08/2015 Page 2 of 2


DATA SHEET 

Reagent: 

pcCCR4 

Catalog 

Number: 

3324 


Release 

Category: 

C 


Description: 



CCR4. The insert was digested with BamHI and SalI and cloned into the BamHI and XhoI 

sites of pcDNAI/amp (SalI and XhoI are compatible ends for ligation). Constructs are 

selected for by adding Ampicillin (100 µg/mL) to the growth medium. Chloramphenicol 

(100 µg/ml) should be added 4.5–5.5 hours (OD 600 = 1.0) after seeding the culture. The 



CCR-4.5'BamHI (33mer): 5'-CGTCGGATCCGCAAGCTGCTTCTGGTTGGGCCC-3'. 

CCR-4.3'SalI (34mer): 5'-CGGCGTGTCGACGAATGTGGAAAAGTTCATTGAC-3'. 



Special 


Plasmid Map 

Recommended 

Storage: 

-20°C 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcCCR4 (Cat #3324) from 








REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pBABE.CCR4 



Release 

Category: 

C 


Description: 



of CCR4. The PCR product was digested with BamHI and SalI and cloned into the 

pBABE-puro. Constructs are selected for by adding Ampicillin (100 µg/mL) to the growth 

medium. Chloramphenicol (100 µg/mL) should be added 4.5–5.5 hours (OD 600 = 1.0) 

after seeding the culture. The plasmid can be transformed into E. coli strain HB101. 


CCR-4.5'BamHI (33mer): 5'-CGTCGGATCCGCAAGCTGCTTCTGGTTGGGCCC-3'. 

CCR-4.3'SalI (34mer): 5'-CGGCGTGTCGACGAATGTGGAAAAGTTCATTGAC-3'. 



Special 


Plasmid Map 

Recommended 

Storage: 

-20°C 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pBABE.CCR4 (Cat #3330) 








REV: 11/08/2015 Page 2 of 2


DATA SHEET 

Reagent: 

pBABE.CCR5 



Release 

Category: 

C 


Description: Chemokine receptor expression vector, CCR5. 



Special 


The cDNA encoding CCR5 was amplified from activated PBMC RNA using primers 

hybridizing to the 5' and 3' untranslated regions of CCR5. The insert was digested with 

BamHI and SalI and cloned into pBABE-puro. Constructs are selected for by adding 

Ampicillin (100 µg/mL) to the growth medium. Chloramphenicol (100 µg/ml) should be 

added 4.5–5.5 hours (OD 600 = 1.0) after seeding the culture. 


CCR-5.5'BamHI (29mer): 5'-CTCGGATCCGGTGGAACAAGATGGATTAT-3'. 

CCR-5.3'SalI (28mer): 5'-CTCGTCGACATGTGCACAACTCTGACTG-3'. 

Plasmid map and sequence file lot 080090 

Contributor provided sequence information 

Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 


with multiple drug selection markers and a complementary helper-free packaging cell 

line. Nucleic Acids Res 18:3587-3596, 1990. 




NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pBABE.CCR5 (Cat 

#3331) from Dr. Nathaniel Landau." Also include the references cited above in any 

publications. 

Patent-pending. Requests from commercial organizations must be directed to 

the New York University Medical Center, ATTN: Office of Industrial Liaison, 550 

First Avenue, New York, NY 10016, TEL: 212-263-8178. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcCCR5 



Release 

Category: 

C 

Provided: 1 mL glycerol stock (E. coli strain, HB101). 

Description: 



of CCR5. The insert was digested with BamHI and SalI and cloned into the BamHI and 

XhoI sites of pcDNAI/amp (SalI and XhoI are compatible ends for ligation). Constructs are 


(100 µg/ml) should be added 4.5–5.5 hours (OD600 = 1.0) after seeding the culture. 


CCR-5.5'BamHI (29mer): 5'-CTCGGATCCGGTGGAACAAGATGGATTAT-3'. 

CCR-5.3'SalI (28mer): 5'-CTCGTCGACATGTGCACAACTCTGACTG-3'. 



Special 


Plasmid Map 

Recommended 

Storage: 

-70°C 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcCCR5 (Cat 


publications. 





Expression Vector Cat. No. 

pc.CCR-1 3321 

pBABE.CCR-1 3327 

pc.CCR-2B 3322 

pBABE.CCR-2B 3328 

pc.CCR-3 3323 


pc.CCR-4 3324 


pc.CCR-5 3325 

pc.Fusin 3326 


pBABE.Fusin 3332 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcCCR5 



Release 

Category: 

C 


Description: Chemokine receptor, CCR5 expression vector. 



Special 


The cDNA encoding CCR5 was amplified from activated PBMC RNA using primers 

hybridizing to the 5' and 3' untranslated regions of CCR5. The insert was digested with 

BamHI and SalI and cloned into the BamHI and XhoI sites of pcDNAI/amp (SalI and XhoI 

are compatible ends for ligation). Constructs are selected for by adding Ampicillin (100 

µg/mL) to the growth medium. Chloramphenicol (100 µg/ml) should be added 4.5–5.5 

hours (OD600 = 1.0) after seeding the culture. 

CCR5 primer sequences: 

CCR5.5'BamHI (29mer): 5'-CTCGGATCCGGTGGAACAAGATGGATTAT-3'. 

CCR5.3'SalI (28mer): 5'-CTCGTCGACATGTGCACAACTCTGACTG-3'. 


Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 



Nucleic Acids Res 18:3587-3596, 1990. Abstract 



of a major co-receptor for primary isolates of HIV-1. Nature 381:661-666, 1996. Abstract 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcCCR5 (Cat 


publications. 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pc.Rh-CCR5 





Description: This expression vector produces rhesus CCR5. 

Cloning Site: 

The insert is cloned into the CMV promoter-driven expression vector pcDNAI/Amp as a 

1.1 kb BamHI-XhoI fragment. 


Special 


Expressed CCR5 can be detected by FACS analysis. 

Other available clones in this set: pcRh-CD4 (Cat #3600), pcRh-CXCR4 (Cat #3601) 



Recommended 

Storage: 

-70°C. 


References: 


immunodeficiency virus use CCR5 as a coreceptor for entry. J Virol 71:2705-2714, 

1997. 




REV: 11/08/2015 Page 1 of 2

NOTE: 





Clone 

Cat. 

No. 

Lot No. 

pc.Rh-CD4 3600 4 99040 


3 97100 4 

99041 

pc.Rh-CCR5 3602 4 99042 

Description 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pBABE.Rh-CCR5 




Provided: 1 ml ampicillin-resistant, transformed TOP-10 bacteria (glycerol stock). 

Description: 

Rhesus CCR5 is cloned into pBABE.puro's BamHI–SalI sites as a BamHI–XhoI 

fragment. 

Cloning Site: BamHI–SalI; Insert size 6.2 Kb. 

Cloning Vector: pBABE.puro 

Special 


Expressed CCR5 can be detected by FACS analysis. The GenBank accession number 

for Rh-CCR5 is U73739. 

Recommended 

Storage: 

-70°C. 


References: 



1997. 

NOTE: 



pBABE.Rh-CCR5 from Dr. Preston Marx." Also include the reference cited above in any 

publications. 




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Chemokine Receptors >> CXCR4

DATA SHEET 

Reagent: 

pBABE.Rh-CXCR4 




Provided: 1 mL ampicillin-resistant, transformed TOP-10 bacteria (glycerol stock). 

Description: 

Rhesus CXCR4 is cloned into pBABE-puro's EcoRI–SalI sites as an EcoRI–XhoI 

fragment. 

Cloning Site: EcoRI–SalI; Insert size 6.2 Kb. 

Cloning Vector: pBABE-puro 

Special 


Expressed CXCR4 can be detected by FACS analysis. The GenBank accession number 

for Rh-CXCR4 is U73740. 

Recommended 

Storage: 

-70°C. 


References: 



1997. 

NOTE: 



pBABE.Rh-CXCR4 from Dr. Preston Marx." Also include the reference cited above in 





REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcCXCR4 



Release 

Category: 

C 

Provided: 1 vial containing 5 μg of purified DNA in TE buffer 

Description: 

CXCR4 expression vector. The cDNA encoding CXCR4 was amplified from activated PBMC 

RNA using primers hybridizing to the 5' and 3' untranslated regions of CXCR4. The insert 

was digested with HindIII and XhoI and cloned into pcDNAI/amp. Constructs are selected 

for by adding Ampicillin (100 µg/mL) to the growth medium. Chloramphenicol (100 

µg/ml) should be added 4.5–5.5 hours (OD600 = 1.0) after seeding the culture. 

CXCR4 primer sequences: 

CXCR4.5'HindIII (30mer): 5'-GGCTAAAGCTTCAAAGTGACGCCGAGGGCC-3'. 

CXCR4.3'XhoI (30mer): 5'-CGTCCTCGAGCATCTGTGTTAGCTGGAGTG-3'. 

Cloning Site: HindIII and XhoI (5'-3'). The size of the insert is approximately 1.3 kb. 


Special 


Alternate name: pcFUSIN 

Plasmid Map 

Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 


with multiple drug selection markers and a complementary helperfree packaging cell 

line. Nucleic Acids Res 18:3587–3596, 1990. 



of a major co-receptor for primary isolates of HIV-1. Nature 381:661–666, 1996. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcCXCR4 (Cat# 

3326)from Dr. Nathaniel Landau." Also include the references cited above in any 

publications. 



First Avenue, New York, NY 10016, Tel: 212-263-8178. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pBABE.CXCR4 



Release 

Category: 

C 

Provided: 1 mL glycerol stock, HB101 cells. 

Description: 

CXCR4 expression vector. The cDNA encoding CXCR4 was amplified from activated PBMC 

RNA using primers hybridizing to the 5' and 3' untranslated regions of CXCR4. The PCR 

product was first cloned into the PCRII vector, digested with EcoRI, and cloned into the 

EcoRI site of pBABE puro. Constructs are selected for by adding Ampicillin (100ug/mL) to 

the growth medium. Chloramphenicol (25 µg/ml) should be added 4.5–5.5 hours (OD 600 

= 1.0) after seeding the culture. 

CXCR4 primer sequences: 

FUSIN.5'(22mer): 5'-AAGTGACGCCGAGGGCCTGAGT-3'. 

FUSIN.3'(22mer): 5'-GCCTAGACACACATCAATATGA-3'. 

Cloning Site: EcoRI. The size of the insert is approximately 1.3 kb. 


Special 


Alternate name: pBABE.FUSIN 

Plasmid Map 

Recommended 

Storage: 

-70°C 





REV: 11/08/2015 Page 1 of 2

References: 


with multiple drug selection markers and a complementary helper-free packaging cell 

line. Nucleic Acids Res 18:3587-3596, 1990. 




NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pBABE.CXCR4 

(Cat# 3332) from Dr. Nathaniel Landau." Also include the references cited above in any 

publications. 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pc.Rh-CXCR4 



4 99041 



Description: This expression vector produces rhesus CXCR4. 

Cloning Site: 

The insert is cloned into the CMV promoter driven expression vector pcDNAI/Amp as a 

1.1 kb EcoRI-XhoI fragment. 


Special 


Expressed CXCR4 can be detected by FACS analysis. 

Other available clones in this set: pcRh-CD4 (Cat #3600), pcRh-CCR5 (Cat #3602) 



Recommended 

Storage: 

-70°C. 


References: 


immunodeficiency virus use CCR5 as a coreceptor for entry. J Virol 71:2705-2714, 

1997. 




REV: 11/08/2015 Page 1 of 2

NOTE: 





Clone 

Cat. 

No. 

Lot No. 

pc.Rh-CD4 3600 4 99040 


3 97100 4 

99041 

pc.Rh-CCR5 3602 4 99042 

Description 










REV: 11/08/2015 Page 2 of 2


DATA SHEET 

Reagent: 

pcCCR1 

Catalog 

Number: 

3321 


Release 

Category: 

C 


Description: 

Chemokine receptor expression vector, CCR1. The cDNA encoding the CCR1 was amplified 


CCR1. The PCR product was digested with BamHI and SalI and cloned into the BamHI and 

XhoI sites of pcDNAI/amp (XhoI and SalI are compatible ends for ligation). Constructs are 


(100 µg/ml) should be added 4.5-5.5 hours (OD 600 = 1.0) after seeding the culture. The 



CCR-1.5'BamHI (34mer): 5'-GCTCAGGATCCGCCCAGAAACAAAGACTTCACGG-3'. 

CCR-1.3'SalI (34mer): 5'-CGATCGGTCGACGTTCTATGTTCCCCAGGATTCC-3'. 


Cloning Vector: pCDNAI/amp (Invitrogen). 

Special 


Plasmid Map 

Recommended 

Storage: 

-20°C 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcCCR1 (Cat# 3321)from 








REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pBABE.CCR1 



Release 

Category: 

C 


Description: 



of CCR1. The PCR product was digested with BamHI and SalI and cloned into pBABE-puro. 

Constructs are selected for by adding Ampicillin (100ug/mL) to the growth medium. 

Chloramphenicol (100 µg/ml) should be added 4.5–5.5 hours (OD 600 = 1.0) after 

seeding the culture. 


CCR-1.5'BamHI (34mer): 5'-GCTCAGGATCCGCCCAGAAACAAAGACTTCACGG-3'. 

CCR-1.3'SalI (34mer): 5'-CGATCGGTCGACGTTCTATGTTCCCCAGGATTCC-3'. 

Plasmid Map 

Cloning Site: BamHI and SalI. The size of the insert is approximately 1.6 kb. 


Recommended 

Storage: 

-20°C 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pBABE.CCR1 (Cat# 3327) 








REV: 11/08/2015 Page 2 of 2

Chemokines >> MIP-1Alpha_MIP-1Beta

DATA SHEET 

Reagent: 

pET32a/Rhesus MIP-1β 


Lot Number: 1 12/9/96 


Provided: 

Plasmid clone transfected into E. coli AD494 (DE3)pLysS as a frozen 30% glycerol 

stock. 

Description: 

Contains the sequence coding for the mature form of rhesus macaque MIP-1b cloned 

into the KpnI-BamHI site of pET32a (Novagen, Madison WI).Rhesus macaque MIP-1ß 

was obtained by RT-PCT of activated rhesus PBMCs. 

Protocol for Purification of Recombinant Rhesus MIP-1α, MIP-1β and RANTES 

Cloning Site: KpnI / BamHI 

Cloning Vector: pET32a (Novagen, Madison WI) 

Special 


This clone contains the sequence coding for the mature form of rhesus macaque 

chemokine MIP-1ß. An amino terminal histidine tag (6x) and S-Tag are linked in frame 

to the recombinant gene (227 aa fusion protein) via an enterokinase cleavage site 

that releases the 69 aa biologically active protein. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Francois Villinger and Dr. Aftab Ansari. 




REV: 11/08/2015 Page 1 of 2

NOTE: 

The E. coli host strain AD494 should not be used for commercial purposes without 

prior consent of the Brookhaven National Laboratories 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pET32a / 

Rhesus MIP-1ß Expression Vector from Dr. Francois Villinger." 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pET32a / Rhesus MIP-1α Expression Vector 




Provided: 

Plasmid clone transfected into E. coli AD494 (DE3) pLysS as a frozen 30% glycerol 

stock. 

Description: 

Contains the sequence coding for the mature form of rhesus macaque MIP-1a cloned 

into the KpnI-BamHI site of pET32a (Novagen, Madison WI). Rhesus macaque MIP-1a 

was obtained by RT-PCT of activated rhesus PBMCs. 

Protocol for the Purification of Recombinant Rhesus MIP-1α, MIP-1β and RANTES 



Special 



chemokine MIP-1a. An amino terminal histidine tag (6x) and S-Tag are linked in frame 

to the recombinant gene (230 aa fusion protein) via an enterokinase cleavage site 

that releases the 72 aa biologically active protein. 

Recommended 

Storage: 

-70°C. 





REV: 11/08/2015 Page 1 of 2

NOTE: 

The E. coli host strain AD494 should not be used for commercial purposes without 

prior consent of the Brookhaven National Laboratories. 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pET32a / 

Rhesus MIP-1a Expression Vector from Dr. Francois Villinger." 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pET32a/Human MIP-1ß Expression Vector 




Provided: 

Plasmid clone transfected into E. coli AD494 (DE3) pLysS as a frozen 30% glycerol 

stock. 

Description: 

Contains the sequence coding for the mature form of human MIP-1b cloned into the 

KpnI-BamHI site of pET32a (Novagen, Madison WI). 

Protocol for the Purification of Recombinant Human MIP-1β 

Cloning Site: KpnI–BamHI. 

Cloning Vector: PET32a (Novagen, Madison WI). 

Special 


This clone contains the sequence coding for the mature form of human MIP-1ß. An 

amino terminal histidine tag (6x) and S-tag are linked in frame to the recombinant 

gene (227 aa) fusion protein) via an enterokinase cleavage site that releases the 69 aa 

biologically active protein. The human MIP-1ß was obtained by RT-PCR from RNA of 

activated human PBMCs. 

Contributor: Dr. Francois Villinger. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pET32a/Human 

MIP-1ß Expression Vector from Dr. Francois Villinger." 

The E. coli host strain AD494 should not be used for commercial purposes without prior 

consent of the Brookhaven National Laboratories. 





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REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pET32a/Mangabey MIP-1β 




Provided: Plasmid clone transfected into E. coli AD494 (DE3) pLysS as a frozen 30% glycerol stock. 

Description: 

Contains the sequence coding for the mature form of mangabey MIP-1b cloned into the 

KpnI-BamHI site of pET32a (Novagen, Madison WI). 

Protocol for the Purification of Recombinant Mangabey MIP-1β 

Cloning Site: KpnI–BamHI. 

Cloning Vector: PET14B (Novagen, Madison WI). 

Special 


This clone contains the sequence coding for the mature form of mangabey chemokine 

macrophage inflammatory protein 1ß. Mangabey MIP-1ß was obtained by RT-PCR from 

RNA of activated mangabey PBMCs. An amino terminal histidine tag (6x) and S-Tag are 

linked in frame to the recombinant gene (227 aa fusion protein) via an enterokinase 

cleavage site that releases the 69 aa biologically active protein. 


References: 

Center DN, Kornfeld H, Cruikshank WW. Interleukin 16 and its function as a CD4 ligand. 

Immunol Today 17:476, 1996. 




REV: 11/08/2015 Page 1 of 2

NOTE: 





pET32a/Mangabey MIP-1ß expression vector from Dr. Francois Villinger." Also include 






REV: 11/08/2015 Page 2 of 2

Chemokines >> Rantes

DATA SHEET 

Reagent: 

pET32a/Rhesus RANTES 




Provided: Plasmid clone transfected into E. coli AD494 (DE3)pLysS as a frozen 30% glycerol stock. 

Description: 

Contains the sequence coding for the mature form of rhesus macaque RANTES cloned 

into the KpnI-BamHI site of pET32a (Novagen, Madison WI). 

Rhesus macaque RANTES was obtained by RT-PCR from RNA of activated rhesus PBMC. 

The sequence information, production and purification protocols are attached. Note: 

There is constraint on the aminoterminus of B-chemokines which is why the insert is 

cloned after an enterokinase cutting site to avoid addition of any extraneous residues. 

The primer used for subcloning encompassed the EK site with the KpnI site. Thus one 

has to use EK in order to obtain a biologically active molecule. 

Protocol for Purification of Recombinant Rhesus MIP-1α, MIP-1β and RANTES 

Sequence 



Special 



chemokine RANTES. An amino terminal histidine tag (6x) and S-Tag are linked in frame 

to the recombinant gene (226 aa fusion protein) via an enterokinase cleavage site that 

releases the 68 aa biologically active protein. 

Recommended 

Storage: 

-70°C. 




REV: 11/08/2015 Page 1 of 2


NOTE: 




through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pET32a / Rhesus 

RANTES Expression Vector from Dr. Francois Villinger." 





REV: 11/08/2015 Page 2 of 2

Chemokines >> SDF-1α

DATA SHEET 

Reagent: 

pQE30T gly-met SDF-1α/CXCL12 



Release 

Category: 

C 

Provided: 2 μg of purified plasmid DNA. 

Description: 

See references 1-3 . The pQE30 plasmid from QIAGEN was modified to contain a tobacco 

etch virus (TEV) protease cleavage site following the hexa-histidine tag and preceding the 

multiple cloning site. The DNA encoding mature SDF-1α was cloned into the BamHI (5′) 

and HindIII (3′) sites. Subsequently, site directed mutagenesis was used to modify the 

TEV protease site from ENLYFQGS to ENLYFQGM. This results in the DNA coding for 

modified TEV protease site preceding the DNA coding for mature SDF-1α. Since mature 

SDF-1α protein has no Met residues, SDF-1α expressed with this vector can be subjected 

to CNBr cleavage to remove the hexa-histidine tag. This leaves mature SDF-1α with a 

native N-terminus. XL-1 blue E. coli should be used for propagation or production of more 

plasmid. SG 13009 E. coli with pREP4 should be used for expression. 

Sequence 

Special 


This clone is unique in that the mature SDF-1α sequence is preceded by a Met residue. 

Since the mature SDF-1α sequence has no Met residues CNBr cleavage can be used to 

generate the native N-terminus of SDF-1α, which is essential for signaling. 

Recommended 

Storage: 

-20°C 

Contributor: Dr. Brian F. Volkman 




REV: 11/08/2015 Page 1 of 2

References: 

Veldkamp, C. T., Peterson, F. C., Hayes, P. L., Mattmiller, J. E., Haugner, J. C., 3rd, de la 

Cruz, N. & Volkman, B. F. (2007). On-column refolding of recombinant chemokines for 

NMR studies and biological assays. Protein Expr. Purif. 52, 202-209. 

Veldkamp, C. T., Seibert, C., Peterson, F. C., Sakmar, T. P. & Volkman, B. F. (2006). 

Recognition of a CXCR4 sulfotyrosine by the chemokine stromal cell-derived factor-1α 

(SDF-1α/CXCL12). J. Mol. Biol. 359, 1400-1409. 

Veldkamp, C. T., Peterson, F. C., Pelzek, A. J. & Volkman, B. F. (2005). The 

monomer-dimer equilibrium of stromal cell-derived factor-1 (CXCL 12) is altered by pH, 

phosphate, sulfate, and heparin. Protein Sci.14, 1071-1081. 

NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH (Cat#11435) from Dr. 

Volkman." Also include the reference cited above in any publications. 


Category C Reagents (Cat #11435) must contact Dr. Brian F. Volkman, 

Department of Biochemistry, Medical College of Wisconsin, 8701 Watertown 

Plank Rd, Milwaukee WI 53202, bvolkman@mcw.edu, before the reagent can be 

released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pQE30T gly-met SDF-1α/CXCL12 



Release 

Category: 

C 

Provided: 1 vial containing 5µg of purified DNA in TE buffer 

Description: 

See references 1-3 . The pQE30 plasmid from QIAGEN was modified to contain a tobacco 

etch virus (TEV) protease cleavage site following the hexa-histidine tag and preceding the 

multiple cloning site. The DNA encoding mature SDF-1α was cloned into the BamHI (5′) 

and HindIII (3′) sites. Subsequently, site directed mutagenesis was used to modify the 

TEV protease site from ENLYFQGS to ENLYFQGM. This results in the DNA coding for 

modified TEV protease site preceding the DNA coding for mature SDF-1α. Since mature 

SDF-1α protein has no Met residues, SDF-1α expressed with this vector can be subjected 

to CNBr cleavage to remove the hexa-histidine tag. This leaves mature SDF-1α with a 

native N-terminus. XL-1 blue E. coli should be used for propagation or production of more 

plasmid. SG 13009 E. coli with pREP4 should be used for expression. 

Sequence 

Special 


This clone is unique in that the mature SDF-1α sequence is preceded by a Met residue. 

Since the mature SDF-1α sequence has no Met residues CNBr cleavage can be used to 

generate the native N-terminus of SDF-1α, which is essential for signaling. 

Recommended 

Storage: 

-20°C 

Contributor: Dr. Brian F. Volkman 




REV: 11/08/2015 Page 1 of 2

References: 

Veldkamp, C. T., Peterson, F. C., Hayes, P. L., Mattmiller, J. E., Haugner, J. C., 3rd, de la 

Cruz, N. & Volkman, B. F. (2007). On-column refolding of recombinant chemokines for 

NMR studies and biological assays. Protein Expr. Purif. 52, 202-209. 

Veldkamp, C. T., Seibert, C., Peterson, F. C., Sakmar, T. P. & Volkman, B. F. (2006). 

Recognition of a CXCR4 sulfotyrosine by the chemokine stromal cell-derived factor-1α 

(SDF-1α/CXCL12). J. Mol. Biol. 359, 1400-1409. 

Veldkamp, C. T., Peterson, F. C., Pelzek, A. J. & Volkman, B. F. (2005). The 

monomer-dimer equilibrium of stromal cell-derived factor-1 (CXCL 12) is altered by pH, 

phosphate, sulfate, and heparin. Protein Sci.14, 1071-1081. 

NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH (Cat#11435) from Dr. 

Volkman." Also include the reference cited above in any publications. 


Category C Reagents (Cat #11435) must contact Dr. Brian F. Volkman, 

Department of Biochemistry, Medical College of Wisconsin, 8701 Watertown 

Plank Rd, Milwaukee WI 53202, bvolkman@mcw.edu, before the reagent can be 

released. 




REV: 11/08/2015 Page 2 of 2

Cytokines >> IL-16

DATA SHEET 

Reagent: 

pET14B/Rhesus IL-16 




Provided: Plasmid clone transfected into E. coli BBL-21 as a frozen 30% glycerol stock. 

Description: 

Contains a fragment coding for the biologically active form of rhesus macaque IL-16 

cloned into the NdeI–BamHI site of pET14B (Novagen, Madison WI). 

Protocol for the Purification of Recombinant MACAQUE IL16 

Special 


Rhesus macaque IL-16 was obtained by RT-PCR from RNA of activated rhesus PBMCs. 

An amino-terminal histidine tag (65) is linked in frame to the recombinant gene via a 

thrombin cleavage site which leaves additional G-S-H residues upstream of the IL-16 

sequence. These amino acids do not seem to affect the biological activity of the 

molecule that is contained within the last 130 aa. 


NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pET14B/Rhesus 

IL-16 Expression Vector from Dr. Francois Villinger." 




REV: 11/08/2015 Page 1 of 1

DC-SIGN-Related >> DC-SIGN

DATA SHEET 

Reagent: 

pcDNA-PtDC-SIGN 



Release 

Category: 

B 

Provided: 1 vial transformed JM109 (glycerol stocks). 

Description: 

The pigtailed macaque DC-SIGN homologue was cloned by reverse transcriptase PCR 

(RT-PCR) using SIGN-1 (5'-AGA GTG GGG TGA CAT GAG TGA CTC-3') and SIGN-END 

(5'-GTG AAG TTC TGC TAC GCA GGA G-3') primers. Both oligo(dT) priming and random 

hexamer priming were used for cDNA synthesis. A portion of each cDNA reaction was used 

for PCR amplification under the following conditions: 94degreeC for 3 min followed by 35 

cycles of 94degreeC for 30 s, 56degreeC for 30 s, and 72degreeC for 2 min. This 

procedure yielded a 1.1-kb fragment from both the oligo(dT)- and random 

hexamer-primed cDNA reactions. Products were subsequently blunt end cloned into 

pBluescript KS(+) and were sequenced. The pigtailed macaque DC-SIGN cDNA was 

excised from pKS+ with EcoRI and SalI and cloned into pcDNA3 using the EcoRI and XhoI 

sites. Consequently the SalI and XhoI sites were destroyed. The sequence has been 

deposited in GenBank under the number AF343727. 

Cloning Site: 

The pigtailed macaque DC-SIGN sequence was inserted via the EcoRI and XhoI sites of 

pcDNA3. The size of the insert is approximately 1.15 kb. 


The cloning vector is pcDNA3 (Invitrogen); the size of the insert plus vector is 

approximately 5.55 kb. 

Special 


DC-SIGN is a C-type lectin, which efficiently binds and transmits HIV-1, HIV-2 and SIV to 

receptor positive cells. Pigtailed macaque DC-SIGN supported HIV-1, HIV-2, and SIV 

binding and transmission and DC-SIGN expression. 

Recommended 

Storage: 

-70°C. 




REV: 11/08/2015 Page 1 of 2

Contributor: Dr. Jason T. Kimata. 

References: Baribaud F, Pohlmann S, Sparwasser T, Kimata MT, Choi YK, Haggarty BS, Ahmad N, 

Macfarlan T, Edwards TG, Leslie GJ, Arnason J, Reinhart TA, Kimata JT, Littman DR, Hoxie 

JA, Doms RW. Functional and antigenic characterization of human, rhesus macaque, 

pigtailed macaque, and murine DC-SIGN. J Virol 75:10281-10289, 2001. 

NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcDNA-PtDC-SIGN from Dr. 

Jason T. Kimata." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3-DC-SIGN 



Release 

Category: 

A 

Provided: 10 µL purified plasmid DNA at 0.7 µg/µL in TE buffer, pH 8.0. 

Description: 

DC-SIGN cDNA was amplified from human dendritic cells and cloned between the BamHI, 

EcoRI restriction sites of pcDNA3. The primers used to amplify DC-SIGN and to introduce 

the necessary restrictions sites were p5-DC 

(5’-CCGGATCCAGAGTGGGGTGACATGAGTG-3’) and p3-DC 

(5’-CCGAATTCGGAAGTTCTGCTACGCAGGAG-3’). The sequence obtained differs at two 

positions from the published sequence. Glu 241 is encoded by GAA instead of GAG and Ser 

333 is encoded by TCG instead of TCA. 

Cloning Site: 

The DC-SIGN sequence (GenBank Accession #M98457) was inserted via BamHI (5’) and 

EcoRI (3’) into pcDNA3. The size of the insert is approximately 1.2 kb. 


The cloning vector is pcDNA3 (Invitrogen), the size of the insert plus vector is 


Special 


DC-SIGN is a C-type lectin, which efficiently binds and transmits HIV-1, HIV-2 and SIV to 

receptor positive cells. This clone can be used for DC-SIGN expression in mammalian 

cells. For example, transfection of 293T cells with this clone results in robust DC-SIGN 

surface expression. Other cell types which could be used for expression are HeLa and 

COS cells. pcDNA3 carries both an ampicillin and neomycin resistance gene. 

Recommended 

Storage: 

-70°C. 

Contributor: Drs. S. Pöhlmann, F. Baribaud, F. Kirchhoff and R.W. Doms. 




REV: 11/08/2015 Page 1 of 2

References: Pöhlmann S, Baribaud F, Lee B, Leslie GJ, Sanchez MD, Hiebenthal-Millow K, Münch J, 

Kirchhoff F, Doms RW. DC-SIGN interactions with human immunodeficiency virus type 1, 

type 2 and simian immunodeficiency virus. J Virol 75:4664-4672, 2001. 

NOTE: 


through the NIH AIDS Reagent Program, NIAID, NIH: pcDNA3-DC-SIGN from Drs. S. 

Pöhlmann, F. Baribaud, F. Kirchhoff and R.W. Doms." Also include the reference cited 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pMX DC-SIGN 




Provided: 1 vial of transformed DH5a bacteria in LB broth containing 20% glycerol. 

Description: 

Contains a ~1.2 kb insert encoding DC-SIGN (GenBank Accession #M98457), cloned 

into the BamHI-SalI site of the MLV retroviral vector pMX. Confers ampicillin resistance. 

Special 


Clone pMX DC-SIGN is a high-efficiency transducing vector for expression of human 

DC-SIGN in mammalian cells. There is no selectable marker, and expression needs to be 

monitored by using anti-DC-SIGN antibody or gp120 binding. 

Recommended 

Storage: 

-80°C. 

Contributor: Mr. Douglas Kwon and Dr. Dan Littman. 

References: 

Onishi M, Kinoshita S, Morikawa Y, Shibuya A, Phillips J, Lanier LL, Gorman DM, Nolan 

GP, Miyajima A, Kitamura T. Applications of retrovirus-mediated expression cloning. Exp 

Hematol 24:324-329, 1996. Geijtenbeek TB, Kwon DS, Torensma R, van Vliet SJ, van 

Duijnhoven GC, Middel J, Cornelissen IL, Nottet HS, KewalRamani VN, Littman DR, 

Figdor CG, van Kooyk Y. DC-SIGN, a dendritic cell-specific HIV-1-binding protein that 

enhances trans-infection of T cells. Cell 100:587-97, 2000. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pMX DC-SIGN 

from Dr. Douglas Kwon and Dr. Dan Littman." 




REV: 11/08/2015 Page 1 of 2




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DC-SIGN-Related >> L-SIGN and SIGNR

DATA SHEET 

Reagent: 

pcDNA3-L-SIGN6 




Provided: 0.5 ml glycerol stocks. 

Description: 

The insert was derived by RT-PCR from human placental mRNA. It contains the full 

coding sequence of L-SIGN cDNA. The clone has ampicillin and neomycin resistance 

markers. The corresponding sequence is available in GenBank: Accession #AF290887 

(nt 39 to 1184). A restriction map of the insert is attached. 

Restriction Enzyme Mapping 

Cloning Site: 

TOPO-Cloning site; orientation allows transcription of the L-SIGN mRNA under the 

CMV promoter. Insert is 1146 nt. 

Cloning Vector: pcDNA3.1/V5/His-TOPO (Invitrogen). Cloning vector including insert is 6669 nt. 

Special 


This clone represents L-SIGN cDNA containing 6 repeats in exon 4 in contrast to a 

more common variant containing 7 repeats. The plasmid clone can be used for 

expression of L-SIGN protein in eukaryotic cells under CMV promoter. 

Host Strain: TOP10 One Shot R cells (Invitrogen). 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Mary Carrington. 




REV: 11/08/2015 Page 1 of 2

References: 

Soilleux EJ, Barten R, Trowsdale J. DC-SIGN; a related gene, DC-SIGNR; and CD23 

form a cluster on 19p13. J Immunol 165(6):2937-2942, 2000. 

Bashirova AA et al. A DC-SIGN related protein is highly expressed on human liver 

sinusoidal endothelial cells and promotes HIV-1 infection. J Exp Med 193(6):671-678, 

2001. 

NOTE: 


through the NIH AIDS Reagent Program, NIAID, NIH: pcDNA3-L-SIGN6 from Dr. Mary 

Carrington." Also include the appropriate references cited above in any publications. 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3-DC-SIGNR 




Provided: 1 ml transformed DH5α (glycerol stocks). 

Description: 

DC-SIGNR (DC-SIGNRelated) is a C-type lectin that binds ICAM-3 and HIV gp120 and 

transmits HIV-1, HIV-2 and SIV to receptor positive cells. DC-SIGNR is expressed on 

sinusoidal endothelial cells in the liver and lymph nodes. 

Cloning Site: 

The complete DC-SIGNR sequence (GenBank Accession #AF245219) was inserted via 

BamHI (5’) and EcoRI (3’) into pcDNA3. The size of the insert is approximately 1.2 kb. 


The cloning vector is pcDNA3 (Invitrogen); the size of the insert plus vector is 


Special 


This clone can be used for DC-SIGNR expression in mammalian cells. Efficient surface 

expression is achieved in 293T cells. 

Recommended 

Storage: 

-70°C. 

Contributor: Drs. Stefan Pöhlmann, Elizabeth Soilleux, Frédéric Baribaud and Robert W. Doms. 

References: 

Pohlmann S, Soilleux EJ, Baribaud F, Leslie GJ, Morris LS, Trowsdale J, Lee B, Coleman 

N, Doms RW. DC-SIGNR, a DC-SIGN homologue expressed in endothelial cells, binds 

to human and simian immunodeficiency viruses and activates infection in trans. Proc 

Natl Acad Sci USA 98:2670-2675, 2001. 




REV: 11/08/2015 Page 1 of 2

NOTE: 



pcDNA3-DC-SIGNR from Drs. Stefan Pöhlmann, Elizabeth Soilleux, Frédéric Baribaud 

and Robert W. Doms." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

EIAV >> Rev

DATA SHEET 

Reagent: 

pRS-ERev 





Description: 

Equine infectious anemia virus rev (ERev) expression vector. Expression of ERev in 

mammalian cells is driven by the RSV promoter. 

Cloning Site: NcoI-XhoI 

Cloning Vector: pBluescript KS+ 

Special 


Constructed by PCR amplification of the rev ORF from the bicistronic tat/rev cDNA 

and inserting it into the expression plasmid pRSPA. 

Recommended 

Storage: 


Contributor: Dr. David Derse. 

References: 

Martarano L, Stephens R, Rice N, Derse D. Equine infectious anemia virus 

trans-regulatory protein Rev controls viral mRNA stability, accumulation, and 

alternative splicing. J Virol 68:3102-3111, 1994. Abstract 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pRS-ERev 

from Dr. David Derse." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

EIAV >> TAT

DATA SHEET 

Reagent: 

pRS-ETatM 


Lot Number: 1 


Provided: 1 vial transformed DH5a (ampicillin-resistant). 

Description: 

Equine infectious anemia virus tat (Etat) expression vector. Sequences cloned into 

pBluescript KS+. Contains the RSV promoter at the SacI site and SV40 polyA 

sequence at the KpnI site. ETat was derived by PCR amplification of cDNA. 

Sequence 

Cloning Site: NcoI-ApaI 

Cloning Vector: pBluescript KS+ 

Special 


Constitutive expression of ETat in mammalian cells is driven by the RSV promoter. 

GenBank No. M30138. 

Contributor: Dr. David Derse. 

References: 

Dorn P, DaSilva L, Martarano L, Derse D. Equine infectious anemia virus tat: insights 

into the structure, function, and evolution of lentivirus trans-activator proteins. J Virol 

64:1616-1624, 1990. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pRS-ETatM 

from Dr. David Derse." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

FIV >> DU

DATA SHEET 

Reagent: 

FIV-34F10 DU 


Lot Number: 9/23/97 


Provided: 1 vial ampicillin-resistant transformed Bu8049 (glycerol stock). 

Description: 

The FIV-34F10 deoxyuridine triphosphate (DU) coding region (nt 3998-4407) was 

amplified by PCR using primers to facilitate directional cloning into the NdeI-EcoRI 

sites of the pUC112Nde vector. 

Cloning Site: Nde I – Eco RI. 

Cloning Vector: pUC112 Nde. 

Special 


This clone expresses high levels of enzymatically active deoxyuridine triphosphatase 

(DU). The protein is suitable for structural, enzymological, and immunological 

studies. The infectious molecular clone FIV-34F10 is also available (Catalog #1236). 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. John H. Elder. 

References: 

Talbott RL, Sparger EE, Lovelace KM, Fitch WM, Pedersen NC, Luciw PA, Elder JH. 

Nucleotide sequence and genomic organization of feline immunodeficiency virus. Proc 


Elder JH, Lerner DL, Hasselkus-Light CS, Fontenot DJ, Hunter E, Luciw PA, Montelaro 

RC, Phillips TR. Distinct subsets of retroviruses encode dUTPase. J Virol 

66:1791-1794, 1992. 

Wagaman PC, Hasselkus-Light CS, Henson M, Lerner DL, Phillips TR, Elder JH. 




REV: 11/08/2015 Page 1 of 2


Molecular cloning and characterization of deoxyuridine triphosphatase from feline 

immunodeficiency virus (FIV). Virology 196:451-457. 

NOTE: 


through the NIH AIDS Reagent Program, NIAID, NIH: FIV-34F10 DU from Dr. John H. 

Elder." Also include the references cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

FIV-34F10 DU 




Provided: 1 vial of 5 µg (1 µg/µl) of purified DNA in TE buffer. 

Description: 

The FIV-34F10 deoxyuridine triphosphate (DU) coding region (nt 3998-4407) was 

amplified by PCR using primers to facilitate directional cloning into the NdeI-EcoRI 

sites of the pUC112Nde vector. 

Cloning Site: Nde I – Eco RI. 

Cloning Vector: pUC112 Nde. 

Special 


This clone expresses high levels of enzymatically active deoxyuridine triphosphatase 

(DU). The protein is suitable for structural, enzymological, and immunological 

studies. The infectious molecular clone FIV-34F10 is also available (Catalog #1236). 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. John H. Elder. 

References: 

Talbott RL, Sparger EE, Lovelace KM, Fitch WM, Pedersen NC, Luciw PA, Elder JH. 

Nucleotide sequence and genomic organization of feline immunodeficiency virus. Proc 


Elder JH, Lerner DL, Hasselkus-Light CS, Fontenot DJ, Hunter E, Luciw PA, Montelaro 

RC, Phillips TR. Distinct subsets of retroviruses encode dUTPase. J Virol 

66:1791-1794, 1992. 





REV: 11/08/2015 Page 1 of 2


Molecular cloning and characterization of deoxyuridine triphosphatase from feline 

immunodeficiency virus (FIV). Virology 196:451-457. 

NOTE: 


through the NIH AIDS Reagent Program, NIAID, NIH: FIV-34F10 DU from Dr. John H. 





REV: 11/08/2015 Page 2 of 2

FIV >> GAG

DATA SHEET 

Reagent: 

FIV-34F10 Gag 




Provided: 1 mL ampicillin-resistant transformed BL21.DE3 (glycerol stock). 

Description: 

The FIV-34F10 gag coding region was amplified by PCR from the start site (nt 630) to 

a region just 3' of the BamHI site at nt 2380. The 3' end was then digested with EcoRV 

at nt 2050 and HcII (in the pUC12NCO vector) and religated to interrupt the PR coding 

region. 

Special 


This clone expresses the unprocessed FIV-34F10 Gag polyprotein, which includes MA, 

CA, NC, and P2, and terminates within PR of Pol. The protein can be used for structural 

and immunological studies. GenBank #M25729. Protein is expressed with or without 

IPTG induction. The infectious molecular clone FIV-34F10 is also available. 

Contributor: Dr. John Elder. 

References: Talbott RL, et al. Proc Natl Acad Sci USA 86:5743, 1989. 

Elder JH, et al. J Virol 67:1869, 1993. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: FIV-34F10 Gag 

from Dr. John Elder." Also include the references cited above in any publications. 




REV: 11/08/2015 Page 1 of 1

FIV >> POL

DATA SHEET 

Reagent: 

FIV-34F10 PR 




Provided: 1 vial containing 5μg of purified DNA in TE buffer 

Description: 

The FIV-34F10 PR coding region was used in conjunction with clonable primers in a 

PCR reaction, then cloned into the NdeI–HindIII region of PT7-7. 

Special 


The PR coding region of FIV-34F10 protease was modified at three sites (G5→I; N55→T; C84→K) 

to produce an autoproteolysis-resistant PR. Protease expression is IPTG inducible. The protease 

will facilitate structural, enzymatic, and drug efficacy studies. GenBank #M25729. The infectious 

molecular clone FIV-34F10 is also available. 


References: Laco GS, et al. J Virol 71:5505, 1997. 

Talbott RL, et al. Proc Natl Acad Sci USA 86:5743, 1989. 


NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: FIV-34F10 PR 





REV: 11/08/2015 Page 1 of 1

DATA SHEET 

Reagent: 

FIV-34F10 RT 




Provided: 1 vial of 5 µg (1 µg/µl) of purified DNA in TE buffer.. 

Description: 

The FIV-34F10 pol region encoding RT/RNase H was PCR-amplified using clonable 

primers, then cloned into the NdeI-SalI sites of pUC112Nde. 

Special 


Due to high copy number, this clone expresses high levels of for structural, 

enzymatically active RT with or without IPTG induction. The protein is suitable 

enzymological, and immunological studies. The infectious molecular clone FIV-34F10 

is also available (Catalog #1236). 





NOTE: 


through the NIH AIDS Reagent Program, NIAID, NIH: FIV-34F10 RT from Dr. John 





REV: 11/08/2015 Page 1 of 1

DATA SHEET 

Reagent: 

FIV-34F10 Pol 




Provided: 1 vial ampicillin-resistant transformed BL21.DE3 (glycerol stock) 

Description: 

The FIV-34F10 pol gene was cloned into the PT7-7 vector as an EcoRI–BglII fragment 

(nt 1875–6460). An adaptor was added between NdeI and EcoRI to put pol in frame 

for translation. 

Cloning Vector: PT7-7, ampicillin resistance. 

Special 


This clone expresses all Pol proteins expressed by FIV PR, including PR, RT, DU, and 

IN. Expression is IPTG inducible. The polyprotein will facilitate structural, enzymatic, 

and drug efficacy studies. GenBank #M25381.1 for the complete FIV genome. The 

infectious molecular clone FIV-34F10 is also available. 





NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: FIV-34F10 Pol 





REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

FIV-34F10 PR 




Provided: 1 vial ampicillin-resistant transformed BL21.DE3 (glycerol stock). 

Description: 

The FIV-34F10 PR coding region was used in conjunction with clonable primers in a 

PCR reaction, then cloned into the NdeI–HindIII region of PT7-7. 

Special 


The PR coding region of FIV-34F10 protease was modified at three sites (G5→I; N55→T; C84→K) 

to produce an autoproteolysis-resistant PR. Protease expression is IPTG inducible. The protease 

will facilitate structural, enzymatic, and drug efficacy studies. GenBank #M25729. The infectious 

molecular clone FIV-34F10 is also available. 


References: Laco GS, et al. J Virol 71:5505, 1997. 

Talbott RL, et al. Proc Natl Acad Sci USA 86:5743, 1989. 


NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: FIV-34F10 PR 





REV: 11/08/2015 Page 1 of 1

DATA SHEET 

Reagent: 

FIV-34F10 RT 




Provided: 1 vial ampicillin-resistant transformed DH5a (glycerol stock). 

Description: 

The FIV-34F10 pol region encoding RT/RNase H was PCR-amplified using clonable 

primers, then cloned into the NdeI-SalI sites of pUC112Nde. 

Special 


Due to high copy number, this clone expresses high levels of for structural, 

enzymatically active RT with or without IPTG induction. The protein is suitable 

enzymological, and immunological studies. The infectious molecular clone FIV-34F10 

is also available (Catalog #1236). 





NOTE: 


through the NIH AIDS Reagent Program, NIAID, NIH: FIV-34F10 RT from Dr. John 





REV: 11/08/2015 Page 1 of 1

DATA SHEET 

Reagent: 

FIV-34F10 Pol 




Provided: 1 vial containing 5 μg of purified DNA in TE buffer. 

Description: 

The FIV-34F10 pol gene was cloned into the PT7-7 vector as an EcoRI–BglII fragment 

(nt 1875–6460). An adaptor was added between NdeI and EcoRI to put pol in frame 

for translation. 

Cloning Vector: PT7-7, ampicillin resistance. 

Special 


This clone expresses all Pol proteins expressed by FIV PR, including PR, RT, DU, and 

IN. Expression is IPTG inducible. The polyprotein will facilitate structural, enzymatic, 

and drug efficacy studies. GenBank #M25381.1 for the complete FIV genome. The 

infectious molecular clone FIV-34F10 is also available. 





NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: FIV-34F10 Pol 





REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

FIV >> REV

DATA SHEET 

Reagent: 

FIV-PPR Rev 




Provided: 1 vial ampicillin-resistant transformed BL21.DE3 (glycerol stock). 

Description: 

cDNA from FIV-PPR was prepared by reverse transcriptase of RNA from PPR-infected 

cells, and used as a target for PCR. Primers were selected to facilitate directional 

cloning of Rev into the BamHI–EcoRI sites of the pRSET B expression vector. 

Special 


This clone expresses functional FIV Rev for use in structural, enzymological, and 

immunological studies. Protein expression is IPTG inducible. The infectious molecular 

clone FIV-PPR is also available. 


References: Phillips TR, et al. J Virol 66:5464, 1992. 

Phillips TR, et al. J Virol 64:4605, 1990. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: FIV-PPR Rev 





REV: 11/08/2015 Page 1 of 1

HIV-1 >> ENV

DATA SHEET 

Reagent: 

p96ZM651gp140-CD5-opt 



Release 

Category: 

C 

Provided: 20 µg purified plasmid DNA precipitated in 1/10 volume of 3M sodium acetate and 3 

volumes of ethanol, total volume is 200 µl. 

Description: 

This expression vector produces gp140 Env protein which is a codon-optimized form of the 

96ZM651.8 clone (Cat# 6173). 

Cloning Site: EcoRI and BamHI cloning site, 5’→3’. The size of the insert is 2316 bp. 


The cloning vector is pcDNA3.1(-) (Invitrogen), the size of the cloning vector including the 

insert is 7720 bp. The clone is ampicillin and neomycin resistant. 

Special 


Derived from the 96ZM651.8 clone (Cat# 6173). For this construct, the env gene was 

artificially reconstructed by substituting original viral codons with those of highly 

expressed human genes (Hass J, et al. Curr Biol 6:315-324, 1996). The original signal 

peptide (aa 1-29) was replaced with the signal peptide from the human CD5 protein 

(MPMGSLQPLATLYLLGMLVASVLA). The env protein was truncated by introducing a stop 

codon right after the transmembrane domain (FAVLSINR). Besides these modifications, 

the env amino acid sequence is unchanged. 

The size of the translated env protein is 707 aa. 

Valuable for studies of protein expression and function, the generation of subtype specific 

immunological reagents, and the production of DNA based subunit vaccines directed 

against a broader spectrum of viruses. 

GenBank accession number: AY181197. 

Table of codon-optimized 96ZM651.8 genes 




REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-80°C. 

Contributor: Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn. 

NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: p96ZM651gp140-CD5-opt 

from Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn." 


must contact Hayes A. Lowe, J.D., UAB Research Foundation, The Office of 

Intellectual Property Management, AB 1120G, 1530 3rd Ave. S, Birmingham AL 

35294-0111, Tel: 205-975-0843 Fax: 205-934-5427, email: halowe@uab.edu, 

before the reagent can be released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

C 



Description: 

This expression vector produces gp120 Env protein which is a codon-optimized form of 

the 96ZM651.8 clone (Cat# 6173). 

Cloning Site: EcoRI and BamHI cloning site, 5'→3'. The size of the insert is 1716 bp. 


The cloning vector is pcDNA3.1(-) (Invitrogen), the size of the cloning vector including 

the insert is 7120 bp. The clone is ampicillin and neomycin resistant. 

Special 



artificially reconstructed by substituting original viral codons with those of highly 

expressed human genes (Hass J, et al. Curr Biol 6:315-324, 1996). The original signal 

peptide (aa 1-29) was replaced with the signal peptide from the human CD5 protein 

(MPMGSLQPLATLYLLGMLVASVLA). The env protein was truncated by introducing a stop 

codon right after the gp120/gp41 cleavage site (RVVEREKR). Besides these modifications, 

the env amino acid sequence is unchanged. 



immunological reagents, and the production of DNA based subunit vaccines directed 

against a broader spectrum of viruses. 






REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-80°C 


NOTE: 



p96ZM651gp120-CD5-opt from Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn." 









REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

p96ZM651gp160-opt 




Provided: 5 µg purified plasmid DNA, 1 µg/µL, in TE. 

Description: 






the insert is 8266 bp. This clone is ampicillin and neomycin resistant. 

Special 


The full-length env gene was artificially reconstructed by substituting original viral 

codons with those of highly expressed human genes (Hass J, et al. Curr Biol 

6:315-324, 1996). The env amino acid sequence is unchanged. Nine base pairs of 

wildtype viral sequence preceding the initiation codon were retained in this synthetic 

gene. 


Valuable for studies of protein expression and function, the generation of subtype 

specific immunological reagents, and the production of DNA based subunit vaccines 

directed against a broader spectrum of viruses. 







REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 



References: 

Gao F, Li Y, Decker JM, Peyerl FW, Bibollet-Ruche F, Rodenburg CM, Chen Y, Shaw DR, 

Allen S, Musonda R, Shaw GM, Zajac AJ, Letvin N, Hahn BH. Codon usage optimization 

of HIV type 1 subtype C gag, pol, env, and nef genes: in vitro expression and immune 

responses in DNA-vaccinated mice. AIDS Res Hum Retroviruses 2003 Sep;19(9):817-23. 

NOTE: 



p96ZM651gp160-opt from Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn." Also 



reagent must contact Hayes A. Lowe, J.D., UAB Research Foundation, The 

Office of Intellectual Property Management, AB 1120G, 1530 3rd Ave. S, 

Birmingham AL 35294-0111, Tel: 205-975-0843 Fax: 205-934-5427, email: 

halowe@uab.edu, before the reagent can be released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 


Catalog 

Number: 

8663 


Release 

Category: 

C 



Description: 






insert is 7648 bp. This clone is ampicillin and neomycin resistant. 

Special 



artificially reconstructed by substituting original viral codons with those of highly expressed 

human genes (Hass J, et al. Curr Biol 6:315-324, 1996). The env amino acid sequence is 

unchanged. Nine base pairs of wildtype viral sequence preceding the initiation codon were 

retained in this synthetic gene. Moreover, the env protein was truncated by introducing a 

stop codon right after the transmembrane domain (FAVLSINR). 



immunological reagents, and the production of DNA based subunit vaccines directed against 

a broader spectrum of viruses. 






REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-80°C 


References: 

Gao F, Li Y, Decker JM, Peyerl FW, Bibollet-Ruche F, Rodenburg CM, Chen Y, Shaw DR, Allen 

S, Musonda R, Shaw GM, Zajac AJ, Letvin N, Hahn BH. Codon usage optimization of HIV 

type 1 subtype C gag, pol, env, and nef genes: in vitro expression and immune responses in 

DNA-vaccinated mice. AIDS Res Hum Retroviruses 2003 Sep;19(9):817-23. 

NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: p96ZM651gp140-opt from 

Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn." Also include the reference cited above in 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 


Catalog 

Number: 

8664 


Release 

Category: 

C 



Description: 







Special 






retained in this synthetic gene. Moreover, the env protein was truncated by introducing a 

stop codon right after the gp120/gp41 cleavage site (RVVEREKR). 










REV: 11/08/2015 Page 1 of 2


Recommended 

Storage: 



References: 





NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: p96ZM651gp120-opt from 

Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn." Also include the reference cited above in 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

p96ZM651gp140-ct-opt 

Catalog 

Number: 

8665 


Release 

Category: 

C 



Description: 







Special 






retained in this synthetic gene. The cytoplasmic tail was truncated by introducing a stop 

codon 16 aa after the transmembrane domain (FQTLIPN). 










REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-80°C. 

Contributor: Drs. Yingying Li, Frederic Bibollet-Ruche, and Beatrice H. Hahn. 

References: 





NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: p96ZM651gp140-ct-opt from 

Drs. Yingying Li, Frederic Bibollet-Ruche, and Beatrice H. Hahn." 









REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pSyngp120IgG 



Release 

Category: 

B 

Provided: 1 vial ampicillin-resistant, transformed JM109. Glycerol stocks. 

Description: 

Encodes JR-FL gp120 fused to the human IgG1Fc gene, under control of the EF1a 

promoter. Contains a synthetic gene that has optimized human codons. Also encodes the 

leader sequence from CD5. Ampicillin resistant. Digestion with BamHI and HindIII yields 

major DNA fragments of 5.88 Kb, 1.71 Kb, and 1.61 Kb. 

Restriction Digest Information 

Special 


Synthetic human immunodeficiency virus type 1 gp120 sequence in which most wild-type 

codons were replaced with codons from highly expressed human genes (syngp120). In 

vitro expression of syngp120 is considerably increased in comparison to that of the 

respective wild-type sequence. In BALB/c mice, DNA immunization with syngp120 

resulted in significantly increased antibody titers and cytotoxic T-lymphocyte reactivity, 

suggesting a direct correlation between expression levels and the immune response. 

Moreover, syngp120 is characterized by rev-independent expression and a low risk of 

recombination with viral sequences. 

Recommended 

Storage: 

-70°C 

Contributor: Dr. Eun-Chung Park and Dr. Brian Seed. 




REV: 11/08/2015 Page 1 of 2

References: 

Haas J, Park E-C, Seed B. Codon usage limitation in the expression of HIV-1 envelope 

glycoprotein. Curr Biol 6:315–324, 1996. 

Andre S, Seed B, Eberle J, Schraut W, Bultmann A, Haas J. Increased immune response 

elicited by DNA vaccination with a synthetic gp120 sequence with optimized codon usage. 

J Virol 72:1497–1503, 1998. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pSyn gp120 IgG 

from Dr. Eun-Chung Park and Dr. Brian Seed." Also include the references cited above in 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pIIIenv3-1 




Provided: 1 ml (6.9 x 10 9 ampicillin-resistant transformed HB101 bacteria). 

Description: 

An HXB2 proviral fragment from nt 5496 (an artificial SalI site) to the 3'-terminal LTR 

was inserted into the SP64 polylinker SalI-XbaI site. The 3' HIV-1 LTR and SP64 

polylinker were inserted in place of the SV40 early promoter and the dhfr gene from 

SV2-dhfr. The Tat-responsive HIV-1 LTR is used to promote expression of HXB2 rev 

and env. 

Plasmid Map 

Cloning Vector: pSV2-dhfr (Subramani S, et al. Mol Cell Biol 1:854, 1981), SalI-Xbal site. 

Special 


The Tat-responsive HIV-1 LTR is used to promote expression of the HIV-1 (HXB2) rev 

and env genes. Induces syncytia when transfected into CD4-positive cell lines 

expressing the tat-III gene. Contains a defective vpu gene (the initiation codon is 

missing). 

Recommended 

Storage: 

70°C. 

Contributor: Dr. Joseph Sodroski. 

References: 

Sodroski J, Goh WC, Rosen C, Campbell K, Haseftine WA. Role of the HTLVIII/LAV 

envelope in syncytium formation and cytopathicity. Nature 322 470-474, 1986. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH:pIIIenv3-1 from 

Dr. Joseph Sodroski." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pDOLHIVenvR 


Lot Number: 2 


Provided: 

1 ml (1.47 x 10 8 ) of transformed bacteria in 67% superbroth, 33% glycerol solution 

(65% glycerol [v/v], 0.1 M MgSO 4 , 250 mM Tris, pH 8.0). 

Description: This construct was designed as a control for pDOLHIVenv (Catalog #324). 

Cloning Vector: pDOL (Korman AJ, et al. Proc Natl Acad Sci USA 84:2150, 1987. 

Neomycin resistant. The antibiotic ß-kanamycin can be substituted for neomycin. Use 

at a final concentration of 12.5 µg/ml ß-kanamycin for liquid culture; use at 25 µg/ml 

for plates. Keep plates wrapped in foil until use as the antibiotic is light-sensitive. 

Digestion with SalI or BamHI may produce extra bands caused by a site prone to 

nicking during digestion. 

Special 


pDOLHIVenvR contains the same SalI-XhoI region (open reading frames for env, tat, 

and rev) as pDOLHIVenv (Cat# 324), however, it has been inserted in the reverse 

direction. Thus no glycoprotein is produced. 

Plasmid Map 

Recommended 

Storage: 

-80°C. 

Contributor: Dr. Eric O. Freed and Dr. Rex Risser. 




REV: 11/08/2015 Page 1 of 2

References: 

Freed EO, Myers DJ, Risser R. Mutational analysis of the cleavage sequence of the 

human immunodeficiency virus type I envelope glycoprotein precursor gp160. J Virol 

63:4670-4675, 1989. 

NOTE: 


through the NIH AIDS Reagent Program, AIDS Program, NIAID, NIH: pDOLHIVenvR 

from Dr. Eric Freed and Dr. Rex Risser." Also include the reference cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pDOLHIVenv 



Release 

Category: 

A 

Provided: 5 μg plasmid DNA (1 μg μl, neomycin resistant). 

Description: 

The SalI-XhoI region of pNL4-3 (Catalog #114) was introduced into the SalI site of pDOL. 

pDOLHIVenv contains the open reading frames for the pNL4-3 env, tat, and rev coding 

regions. Expression is from the Moloney murine virus LTR. 

Plasmid Map 

Cloning Site: SalI. 

Cloning Vector: pDOL (Korman AJ, et al.Proc Natl Acad Sci USA 84:2150, 1987). 

Special 


The antibiotic ß-kanamycin can be substituted for neomycin. Use at a final concentration 

of 12.5 µg/ml ß-kanamycin for liquid culture; use at 25 µg/ml for plates. Keep plates 

wrapped in foil until use as the antibiotic is light-sensitive. This construct efficiently 

expresses envelope glycoproteins when transfected into HeLa T4 cells. These transfected 

cells form syncytia indistinguishable from those formed by HeLa T4 infected with HIV-1. 

Protein can be detected by immunoprecipitation and immunofluorescence. Digestion with 

SalI or BamHI may produce extra bands caused by a site prone to nicking during digestion. 

Recommended 

Storage: 

-70°C. 





REV: 11/08/2015 Page 1 of 2

References: 

Freed EO, Myers DJ, Risser R. Mutational analysis of the cleavage sequence of the human 

immunodeficiency virus type 1 envelope glycoprotein precursor gp160. J Virol 

63:4670-4675, 1989. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pDOLHIVenv from 

Dr. Eric Freed and Dr. Rex Risser." Also include the reference cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

HIV-1 clone BaL.01 




Provided: 5μg purified plasmid DNA at 1μg/μL 

Description: 

A PCR fragment containing full-length env and rev genes was derived from the genomic 

DNA of infected PBMC. The original virus (HIV-1 BaL) was obtained from NIH AIDS 

Research & Reference Reagent Program (cat #510). The env/rev cassette was cloned 

into pcDNA3.1D/V5-His TOPO © expression vector by a directional cloning approach. A 

single transformed ampicillin resistant E. coli colony was selected and expanded. 

Recombinant plasmid carries resistance genes for ampicillin and neomycin. 

Cloning Site: 

The HIV-1 env/rev cassette was cloned directly into the cloning site of pcDNA3.1D/V5-His 

TOPO © expression vector, in the correct orientation with the CMV promoter. The size of 


Cloning Vector: pcDNA3.1D/V5-His TOPO © 

Special 


The BaL.01 clone expresses a functional env/rev cassette and can be used to generate 

pseudotyped infectious virions. BaL.01 differs from BaL.26 (cat # 11446) by 16 amino 

acid positions in gp160. Ascession #DQ318210 

Sequence 

Recommended 

Storage: 

-20°C or lower 

Contributor: Dr. John Mascola 




REV: 11/08/2015 Page 1 of 2

References: 

Li Y, Svehla K, Mathy NL, Voss G, Mascola JR, Wyatt R. Characterization of antibody 

responses elicited by human immunodeficiency virus type 1 primary isolate trimeric and 

monomeric envelope glycoproteins in selected adjuvants. J. Virology, 80 (3); 1414-26 

(2006). 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HIV-1 clone 

BaL.01 (Cat#11445) from Dr. Mascola." Also include the reference cited above in any 

publications. 





reagent. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 





Provided: 5 µg purified plasmid DNA in TE buffer at 1 mg/mL. 

Description: 



Cloning Site: EcoRI and BamHI cloning site, 5’→3’. The size of the insert is 2719 bp. 




Special 


The envelope was derived from the 96ZM651.8 clone (Cat# 6173). This construct was 

codon-optimized for expression in human cells. The original signal peptide was replaced 

with that of the human CD5 protein (MPMGSLQPLATLYLLGMLVASVLA). The 

codon-optimized translated Env amino acid sequence is unchanged from the original 

viral amino acid sequence. 

The size of the translated Env protein is 863 aa. 




GenBank: AY181197 






REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 



References: 

Haas J, Park EC, Seed B. Curr Biol. 1996 Mar 1;6(3):315-24. Codon usage limitation in 

the expression of HIV-1 envelope glycoprotein. Abstract 

NOTE: 



p96ZM651gp160-CD5-opt from Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn." 


reagent must contact Hayes A. Lowe, J.D., UAB Research Foundation, The Office 

of Intellectual Property Management, AB 1120G, 1530 3rd Ave. S, Birmingham 

AL 35294-0111, Tel: 205-975-0843 Fax: 205-934-5427, email: 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA 89.6 env 




Provided: 1 vial of 0.5 μg of purified DNA at 25 ng/μl in TE buffer. 

Description: 

This plasmid will constitutively express 89.6 env from a CMV promotor. It can be used 

to pseudotype molecular clones missing env. 

A 3,180 bp HindIII-EcoRV fragment from p89.6 (cat# 3552) containing the env open 

reading frame was cloned into the HindIII and EcoRV sites of pcDNA3.1+. 

Cloning Site: 

Digestion with HindIII and EcoRV will release the 3,180 bp insert from the 5.4 kb 

vector. 

Cloning Vector: pcDNA3.1+ (Amp resistant) 

Special 


GenBank U39362 (wild type 89.6) 

The sequence file and map for this construct is HERE. 

Recommended 

Storage: 

-20°C 

Contributor: Drs. Kathleen Collins and Ronald Collman 

References: 

Carter CC, Onafuwa-Nuga A, McNamara LA, Riddell J 4th, Bixby D, Savona MR, Collins 

KL. HIV-1 infects multipotent progenitor cells causing cell death and establishing latent 

cellular reservoirs. Nat Med. 2010 Apr;16(4):446-51. doi: 10.1038/nm.2109. Epub 

2010 Mar 7. PubMed PMID: 20208541; PubMed Central PMCID: PMC2892382. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: Cat# 12845 

pcDNA 89.6 env from Dr. Kathleen Collins and Dr. Ronald Collman." Also include the 

reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pDOLHIVenv 




Provided: 5 μg plasmid DNA (0.228 µg/µl) in TE buffer. 

Description: This construct efficiently expresses pNL4-3 envelope glycoprotein. 

Cloning Site: SalI. 


The MuLV-based retrovirus vector pDOL (Korman AJ, et al.Proc Natl Acad Sci USA 

84:2150, 1987). 

Neomycin resistant. The antibiotic ß-kanamycin can be substituted for neomycin. Use 

at a final concentration of 12.5 µg/ml ß-kanamycin for liquid culture; use at 25 µg/ml 

for plates. Keep plates wrapped in foil until use as the antibiotic is light-sensitive. 

Digestion with SalI or BamHI may produce extra bands caused by a site prone to 

nicking during digestion. 

Special 


The SalI-XhoI region of pNL4-3 (Catalog #114) was introduced into the SalI site of 

pDOL. pDOLHIVenv contains the open reading frames for the pNL4-3 env, tat, and 

rev coding regions. Expression is from the Moloney murine virus LTR. 

Plasmid Map 

Recommended 

Storage: 

-80°C. 





REV: 11/08/2015 Page 1 of 2

References: 

Freed EO, Myers DJ, Risser R. Mutational analysis of the cleavage sequence of the 

human immunodeficiency virus type 1 envelope glycoprotein precursor gp160. J Virol 

63:4670-4675, 1989. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pDOLHIVenv 

from Dr. Eric Freed and Dr. Rex Risser." Also include the reference cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 





Provided: 5 µg of purified DNA at 0.175 µg/µl in TE buffer. 

Description: 

This construct bears a PCR fragment containing full-length env and rev genes from 

HIV-1 BaL (Cat# 510) derived from the genomic DNA of infected PBMC. 

Cloning Site: 

The HIV-1 env/rev cassette was cloned directly into the cloning site of 

pcDNA3.1D/V5-His TOPO© expression vector, in the correct orientation with the CMV 

promoter. The size of the insert is 2559 bp. 


pcDNA3.1D/V5-His TOPO©. The plasmid carries resistance genes for ampicillin and 

neomycin. 

Special 


Accession Number: DQ318211 

The BaL.26 clone expresses a functional env/rev cassette and can be used to 

generate pseudotyped infectious virions. BaL.26 differs from BaL.01 (Cat# 11445) by 

16 amino acid positions in gp160. 

Recommended 

Storage: 

-20°C or lower. 





REV: 11/08/2015 Page 1 of 2

References: 


responses elicited by human immunodeficiency virus type 1 primary isolate trimeric 

and monomeric envelope glycoproteins in selected adjuvants. J. Virology, 80 (3); 

1414-26 (2006). 

NOTE: 




publications. 




released. Please specify the name and a description of the intended use of 

the reagent. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pHXB2-env 





Description: 

HIV-1 gp160 is expressed from an SV40 promoter. No other HIV gene products are 

expressed. 

Cloning Site: 5' SmaI - 3' SalI 

Cloning Vector: pSV7d, Ampicillin resistant 

Special 


Contains a 2897 bp 5' SacI - 3' XhoI HXB2 env fragment from reagent #1067 

HIV-gpt (env coding sequences are nt 6224 - 8794). The 5' SacI insert site was filled 

in and fused to the pSV7d SmaI site. The 3' XhoI insert site was ligated to the SalI 

site of pSV7d. 

SV40 origin provides high levels of gp160 expression in COS cells. Expression is 

rev-dependent and transient. This expression vector has been used with HIV-gpt 

(catalog #1067) to cotransfect COS cells, producing infectious HIV virions. 

Source of Provirus: HIV-1 plasmid pHXB2gpt (Dr. A. Fisher and Dr. F. Wong-Staal). 



Recommended 

Storage: 





REV: 11/08/2015 Page 1 of 2

Contributor: Dr. Kathleen Page and Dr. Dan Littman. 

References: 

Page KA, Landau NR, Littman DR. Construction and use of a human immunodeficiency 

virus vector for analysis of virus infectivity. J Virol 64:5270-5276, 1990. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pHXB2-env 

from Dr. Kathleen Page and Dr. Dan Littman." Also include the reference cited above 

in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pS15-GFP 




Provided: 5 µg at 0.6 mg/mL of purified DNA in TE buffer. 

Description: 

An expression vector that produces a fusion protein between green fluorescent 

protein (GFP) and the N-terminal 15 amino acid sequence of c-Src (S15). The 

protein is efficiently packaged into HIV virions. 

Cloning Vector: Clontech pEGFP 

Recommended 

Storage: 

-20°C for long term storage. 

Contributor: Thomas J. Hope 

NOTE: 

Acknowledgment for publications should read "pS15-GFP cat#12480 was obtained 

through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH from Dr. 

Thomas J. Hope." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 1 of 1

DATA SHEET 

Reagent: 

pS15-mCherry 





Description: 

An expression vector that produces a fusion protein between mCherry and the 

N-terminal 15 amino acid sequence of c-Src (S15). The protein is efficiently packaged 

into HIV virions. 

Cloning Vector: Clontech pmCherry 

Recommended 

Storage: 



References: 

Campbell EM, Perez O, Melar M, Hope TJ. Labeling HIV-1 virions with two fluorescent 

proteins allows identification of virions that have productively entered the target cell. 

Virology. 2007 Apr 10;360(2):286-93. 

NOTE: 

Acknowledgment for publications should read "pS15-mCherry cat#12481 was 

obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH from 

Dr. Thomas J. Hope." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 1 of 1

DATA SHEET 

Reagent: 

pSyngp140JR-FL 



Release 

Category: 

B 

Provided: 1 vial ampicillin-resistant, transformed JM109. 

Description: 

Encodes JR-FL gp140 (env gene minus the transmembrane and C-terminal sequences), 

under control of the EF1a promoter. Contains a synthetic gene that has optimized human 

codons. Also encodes the leader sequence from CD5. Digestion with BamHI and HindIII 

yields major DNA fragments of 5.88 Kb, 2.18 Kb, and 1.71 Kb. 


Special 



codons were replaced with codons from highly expressed human genes. In vitro 

expression of syngp140 is considerably increased in comparison to that of the respective 

wild-type sequence. In BALB/c mice, DNA immunization with syngp140 resulted in 

significantly increased antibody titers and cytotoxic T-lymphocyte reactivity, suggesting a 

direct correlation between expression levels and the immune response. Moreover, 

syngp140 is characterized by rev-independent expression and a low risk of recombination 

with viral sequences. 

Recommended 

Storage: 

-70°C 





REV: 11/08/2015 Page 1 of 2

References: 





J Virol 72:1497–1503, 1998. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pSyn gp140 JR-FL 






REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pCAGGS SF162 gp160 




Provided: 5 μL purified plasmid DNA (1.2 μg/μl). 

Description: 

The SF162 gp160 insert was cut out of a SF162 3’ half-genome clone as a 3.3 kb 

EcoRI-BgIII fragment (nt 1-3287) and cloned into the pCAGGS vector at the same sites 

(Please note that the original BgIII cloning site is no longer present in this construct). 

The plasmid has a CMV enhancer and a chicken b-actin promoter as well as an SV40 

origin. It is ampicillin resistant. 

Click here to view sequence 

Cloning Site: 5′- EcoRI- BgIII -3′. The size of the insert is 3.3 kb. 

Cloning Vector: The cloning vector is pCAGGS (4.8 kb). 

Special 


Co-transfection of 293T cells with NL4-3 env plasmid yields an HIV-1 pseudotype virus 

that is R5-tropic and susceptible to neutralization by multiple antibodies. 

GenBank Accession #: EU123924 

Recommended 

Storage: 

4°C 

Contributor: Drs. Leonidas Stamatatos and Cecilia Cheng-Mayer. 




REV: 11/08/2015 Page 1 of 2

References: 

Stamatatos L., Lim M. and Cheng-Mayer C. Generation and structural analysis of soluble 

oligomeric envelope proteins derived from neutralization-resistant and 

neutralization-susceptible primary HIV-1 isolates. AIDS Res. And Hum. Retroviruses 

16: 981-994, 2000. 

Stamatatos L., Wiskerchen M. and Cheng-Mayer C. Effect of major deletions in the V1 

and V2 loops of a macrophage-tropic HIV-1 isolate on viral envelope structure, 

cell-entry and replication. AIDS Res. And Hum. Retroviruses 14: 1129-1139, 1998. 

Cheng-Mayer C., Liu R., Landau N. R. and Stamatatos L. Macrophage tropism of human 

immunodeficiency virus type 1 and utilization of the CC- CKR5 coreceptor. J. Virol. 

71:1657-1661, 1997. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pCAGGS SF162 

gp160 from Drs. L. Stamatatos and C. Cheng-Mayer." Also include the reference cited 






REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

HIV-1 (YU2) gp140 (-/GCN4) in pcDNA3.1 



Release 

Category: 

C 

Provided: 5 µg of 1.7 µg/µl purified plasmid DNA in TE pH8.0. 

Description: 

Once transfected into appropriate cells, the plasmid produces soluble, cleavage negative, 

trimeric gp140 derived from the neutralization resistant YU-2 virus. Trimerization is due 

to the inclusion of the GCN4 trimeric motif (MKQIEDKIEEILSKIYHIENEIARIKKLIGEV) which 

is positioned after lysine 675 in the gp140 delta675(-/GCN4) construct. Purification of the 

protein is possible through use of a Ni column. Please see the below references for 

additional information. 

Cloning Site: NheI (5’) / AflII (3’) 


The cloning vector is pcDNA3.1(Zeo/-) (Invitrogen), the size of the cloning vector 

including the insert is 7372 bp. 

Recommended 

Storage: 


Contributor: Dr. Joseph Sodroski 

References: 

Yang X, Wyatt R, Sodroski J. Improved elicitation of neutralizing antibodies against 

primary human immunodeficiency viruses by soluble stabilized envelope glycoprotein 

trimers. J Virol. 2001 Feb;75(3):1165-71. DOI: 10.1128/JVI.75.3.1165-1171.2001. Li Y, 

Svehla K, Mathy NL, Voss G, Mascola JR, et al. Characterization of antibody responses 

elicited by human immunodeficiency virus type 1 primary isolate trimeric and monomeric 

envelope glycoproteins in selected adjuvants. J Virol, 2006; 80:1414–1426 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HIV-1 (YU2) gp140 

(-/GCN4) in pcDNA3.1 from Dr. Joseph Sodroski." 


must contact Joseph Sodroski, phone: 617.632.3371, email: 

joseph_sodroski@dfci.harvard.edu, before the reagent can be released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pSyngp120JR-FL 



Release 

Category: 

B 

Provided: 1 vial of purified DNA with 5 μg/vial at 1 μg/μL. 

Description: 

Encodes JR-FL gp120 under control of the EF1a promoter. Also encodes the leader 

sequence from CD5. Digestion with BamHI and HindIII yields major DNA fragments of 

5.88 Kb, 1.71 Kb, and 1.61 Kb. 


Cloning Vector: CDM7-derived vector. Amp resistant. 

Special 



codons were replaced with codons from highly expressed human genes (syngp120). In 

vitro expression of syngp120 is considerably increased in comparison to that of the 

respective wild-type sequence. In BALB/c mice, DNA immunization with syngp120 

resulted in significantly increased antibody titers and cytotoxic T-lymphocyte reactivity, 

suggesting a direct correlation between expression levels and the immune response. 

Moreover, syngp120 is characterized by rev-independent expression and a low risk of 

recombination with viral sequences. 

Recommended 

Storage: 

-70°C 





REV: 11/08/2015 Page 1 of 2

References: 





J Virol 72:1497–1503, 1998. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pSyn gp120 JR-FL 






REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

Resurfaced stabilized core (RSC3) expression vector 



Release 

Category: 

C 

Provided: 

1 vial of 5 μg of purified DNA in 63 μL (0.08 mg/mL) of TE buffer (10 mM Tris, 1 mM 

EDTA, pH8.0). 

Description: 

This expression vector produces the protein RSC3 (cat# 12042). This protein was 

designed to preserve the antigenic structure of the CD4 binding site of gp120 while 

removing other antigenic regions. Resurfaced Stabilized Core 3 (RSC3) is based on the 

HXB2 Ds12F123 stabilized core. The variable regions 1 - 3 are removed. For more details, 

please see Wu et al. 2010. 

The size of the insert is about 1000 base pairs. The size of the cloning vector including 

the insert is 5503 base pairs. The expression vector contains a leader sequence, RSC3, 

Avi-tag, and a 6xHis tag driven by a CMV promoter. 

12279 Plasmid Sequence 

12279 Plasmid Map 

Cloning Site: 

Please see plasmid map for cloning site; orientation is 5’ to 3’. The size of the insert is 

989 base pairs. 

Cloning Vector: CMV8x/R 

Recommended 

Storage: 

-20 o C or less 

Contributor: Zhi-Yong Yang, Gary Nabel, and John Mascola 




REV: 11/08/2015 Page 1 of 2

References: Wu X, Yang ZY, Li Y, Hogerkorp CM, Schief WR, Seaman MS, Zhou T, Schmidt SD, Wu L, 

Xu L, Longo NS, McKee K, O'Dell S, Louder MK, Wycuff DL, Feng Y, Nason M, Doria-Rose 

N, Connors M, Kwong PD, Roederer M, Wyatt RT, Nabel GJ, Mascola JR. Rational design of 

envelope identifies broadly neutralizing human monoclonal antibodies to HIV-1. Science. 

2010 Aug 13;329(5993):856-61.Abstract 

Wu X, Zhou T, Zhu J, Zhang B, Georgiev I, Wang C, Chen X, Longo NS, Louder M, McKee 

K, O'Dell S, Perfetto S, Schmidt SD, Shi W, Wu L, Yang Y, Yang ZY, Yang Z, Zhang Z, 

Bonsignori M, Crump JA, Kapiga SH, Sam NE, Haynes BF, Simek M, Burton DR, Koff WC, 

Doria-Rose NA, Connors M; NISC Comparative Sequencing Program, Mullikin JC, Nabel 

GJ, Roederer M, Shapiro L, Kwong PD, Mascola JR. Focused evolution of HIV-1 

neutralizing antibodies revealed by structures and deep sequencing. Science. 2011 Sep 

16;333(6049):1593-602. Abstract 

(Reference for CMV/R plasmid): Barouch DH, Yang ZY, Kong W-P, Korioth-Schmitz B, 

Sumida SM, Truitt DM, Kishko MG, Arthur JC, Miura A, Mascola JR, Letvin NL, Nabel GJ. A 

human T-cell leukemia virus type 1 regulatory element enhances the immunogenicity of 

human immunodeficiency virus type 1 DNA vaccines in mice and nonhuman primates. J. 

Virol. 2005 Jul;79(14):8828-34 Abstract 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: RSC3 expression 

vector (Cat #12279) from Drs. Zhi-Yong Yang, Gary Nabel, and John Mascola.” Also 

include the references cited above in any publications. 


must contact: The Office of Technology Development, NIAID, 6610 Rockledge 

Drive, Suite 2800, MSC 6606, Bethesda, MD, 20892-6606, Tel: 301-496-2644, 

Fax: 301-402-7123, before the reagent can be released. 

Strictly limited to 2 aliquots per year. Larger requests require contributor 

approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

Resurfaced stabilized core Δ371I (RSC3 Δ371I) expression vector 



Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 

This expression vector (RSC3-Δ371I-His-Avi) produces the protein RSC3 Δ371I (cat# 

12043). This protein was designed to preserve the antigenic structure of the CD4 binding 

site of gp120 while removing other antigenic regions. Resurfaced Stabilized Core 3 (RSC3) 

is based on the HXB2 Ds12F123 stabilized core. The variable regions 1 - 3 are removed. 

The mutation bore on RSC3 Δ371I greatly diminishes the binding of CD4bs mAbs to the 

protein. For more details, please see Wu et al. 2010. 






Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2





2010 Aug 13;329(5993):856-61.Abstract 







16;333(6049):1593-602. Abstract 






NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: RSC3 Δ371I 

expression vector (Cat #12280) from Drs. Zhi-Yong Yang, Gary Nabel, and John Mascola.” 

Also include the references cited above in any publications. 






approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

Resurfaced stabilized core 3 G367R (RSC3 G367R) expression vector 



Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 

This expression vector (RSC3-G367R-His-Avi plasmid) produces the protein RSC3 G367R 

(cat# 12363). This protein was designed to preserve the antigenic structure of the CD4 

binding site of gp120 while removing other antigenic regions. Resurfaced Stabilized Core 3 

(RSC3) is based on the HXB2 Ds12F123 stabilized core. The variable regions 1 - 3 are 

removed. The mutation bore on RSC3/G367R greatly diminishes the binding of CD4bs 

mAbs such as b12 and b13 to the protein. For more details, please see Lynch et al. 2012. 


the insert is 5503 base pairs. The expression vector contains a leader sequence, 

RSC3/G367R, Avi-tag, and a 6xHis tag driven by a CMV promoter. 



Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 

Lynch RM, Tran L, Louder MK, Schmidt SD, Cohen M; CHAVI 001 Clinical Team Members, 

Dersimonian R, Euler Z, Gray ES, Abdool Karim S, Kirchherr J, Montefiori DC, Sibeko S, 

Soderberg K, Tomaras G, Yang ZY, Nabel GJ, Schuitemaker H, Morris L, Haynes BF, 

Mascola JR. The Development of CD4 Binding Site Antibodies during HIV-1 Infection. J 

Virol. 2012 Jul;86(14):7588-95. Abstract 

(References for RSC3) Wu X, Yang ZY, Li Y, Hogerkorp CM, Schief WR, Seaman MS, Zhou 

T, Schmidt SD, Wu L, Xu L, Longo NS, McKee K, O'Dell S, Louder MK, Wycuff DL, Feng Y, 

Nason M, Doria-Rose N, Connors M, Kwong PD, Roederer M, Wyatt RT, Nabel GJ, Mascola 

JR. Rational design of envelope identifies broadly neutralizing human monoclonal 

antibodies to HIV-1. Science. 2010 Aug 13;329(5993):856-61.Abstract 







16;333(6049):1593-602. Abstract 






NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: RSC3 G367R 








approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

Resurfaced stabilized core 3 Δ371I/P363N (RSC3 Δ371I/P363N) expression vector 



Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 

This expression vector (RSC3 Δ371I/P363N-His-Avi plasmid) produces the protein RSC3 

Δ371I/P363N (cat# 12362). This protein was designed to preserve the antigenic structure 

of the CD4 binding site of gp120 while removing other antigenic regions. Resurfaced 

Stabilized Core 3 (RSC3) is based on the HXB2 Ds12F123 stabilized core. The variable 

regions 1 - 3 are removed. The mutation bore on RSC3 Δ371I/P363N was designed to 

abolish the CD4bs of the protein. For more details, please see Lynch et al. 2012. 






Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 

















16;333(6049):1593-602. Abstract 






NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: RSC3 

Δ371I/P363N expression vector (Cat #12281) from Drs. Zhi-Yong Yang, Gary Nabel, and 

John Mascola.” Also include the references cited above in any publications. 






approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 











Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2





2010 Aug 13;329(5993):856-61.Abstract 







16;333(6049):1593-602. Abstract 






NOTE: 










approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 











Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2





2010 Aug 13;329(5993):856-61.Abstract 







16;333(6049):1593-602. Abstract 






NOTE: 










approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 












Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2





2010 Aug 13;329(5993):856-61.Abstract 







16;333(6049):1593-602. Abstract 






NOTE: 










approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 












Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2





2010 Aug 13;329(5993):856-61.Abstract 







16;333(6049):1593-602. Abstract 






NOTE: 










approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 












Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 

















16;333(6049):1593-602. Abstract 






NOTE: 










approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 












Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 

















16;333(6049):1593-602. Abstract 






NOTE: 










approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 












Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 

















16;333(6049):1593-602. Abstract 






NOTE: 










approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

C 

Provided: 


EDTA, pH8.0). 

Description: 












Cloning Site: 




Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 

















16;333(6049):1593-602. Abstract 






NOTE: 










approval. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 





Provided: 5 μg purified plasmid DNA at 1 μg/μL in TE buffer. 

Description: 

This construct bears a PCR fragment containing full-length env and rev genes from 

HIV-1 BaL (Cat# 510) derived from the genomic DNA of infected PBMC. 

Cloning Site: 

The HIV-1 env/rev cassette was cloned directly into the cloning site of 

pcDNA3.1D/V5-His TOPO © expression vector, in the correct orientation with the CMV 

promoter. The size of the insert is 2559 bp. 


pcDNA3.1D/V5-His TOPO © . The plasmid carries resistance genes for ampicillin and 

neomycin. 

Special 


The BaL.01 clone expresses a functional env/rev cassette and can be used to 

generate pseudotyped infectious virions. BaL.01 differs from BaL.26 (Cat# 11446) by 

16 amino acid positions in gp160. Ascension #DQ318210 

Sequence 

Recommended 

Storage: 

-20°C or lower 





REV: 11/08/2015 Page 1 of 2

References: 


responses elicited by human immunodeficiency virus type 1 primary isolate trimeric 

and monomeric envelope glycoproteins in selected adjuvants. J. Virology, 80 (3); 

1414-26 (2006). 

NOTE: 




publications. 





the reagent. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 





Provided: 10μl/vial purified plasmid DNA at 0.8μg/μl. 

Description: 

A PCR fragment containing full-length env and rev genes was derived from the genomic 

DNA of infected PBMC. The original virus (HIV-1 BaL) was obtained from NIH AIDS 

Research & Reference Reagent Program (cat #510). The env/rev cassette was cloned 

into pcDNA3.1D/V5-His TOPO© expression vector by a directional cloning approach. A 

single transformed ampicillin resistant E. coli colony was selected and expanded. 

Recombinant plasmid carries resistance genes for ampicillin and neomycin. 

Cloning Site: 

The HIV-1 env/rev cassette was cloned directly into the cloning site of pcDNA3.1D/V5-His 

TOPO© expression vector, in the correct orientation with the CMV promoter. The size of 


Cloning Vector: pcDNA3.1D/V5-His TOPO©. 

Special 


Accession Number: DQ318211 

The BaL.26 clone expresses a functional env/rev cassette and can be used to generate 

pseudotyped infectious virions. BaL.26 differs from BaL.01 (cat # 11445) by 16 amino 

acid positions in gp160. 

Recommended 

Storage: 

-20°C or lower. 





REV: 11/08/2015 Page 1 of 2

References: 


responses elicited by human immunodeficiency virus type 1 primary isolate trimeric and 

monomeric envelope glycoproteins in selected adjuvants. J. Virology, 80 (3); 1414-26 

(2006). 

NOTE: 




publications. 





reagent. 




REV: 11/08/2015 Page 2 of 2

HIV-1 >> GAG

DATA SHEET 

Reagent: 

p96ZM651gag-opt 




Provided: 19 µg purified plasmid DNA at a concentration of 1 µg/µl in TE. 

Description: 

This expression vector produces the Gag pre-protein which is a codon-optimized form of 






Special 


Derived from the 96ZM651.8 clone (Cat# 6173). For this construct, the full-length gag 

gene was artificially reconstructed by substituting original viral codons with those of 

highly expressed human genes (Hass J, et al. Curr Biol 6:315-324, 1996). The gag 

amino acid sequence is unchanged. Nine base pairs of wildtype viral sequence preceding 

the initiation codon were retained in this synthetic gene. 

The size of the translated gag protein is 494 aa. 










REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 



References: 





NOTE: 



p96ZM651gag-opt from Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn." Also include 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pGag-EGFP 



Release 

Category: 

C 


Description: 

This cDNA directs Rev-independent expression of an HIV-1 Gag-EGFP fusion protein. 

Gag-EGFP forms virus-like particles with an efficiency equivalent to that of Gag. 

Cloning Site: KpnI/BamHI 


pEGFP-NI (Kan R ). The size of the insert is 1530 bp. The size of the vector (including 

insert) is 6274 bp. 

Special 


The insert consists of Rev-independent HIV-1 Gag coding sequences (pCMV55M1-10 

obtained from Dr. George Pavlakis, NCI) fused in frame to EGFP. The clone was 

constructed as follows: 1) Rev independent Gag was amplified by PCR. The 5’ primer 

contained a Kpn I site and overlapped with the start of the coding region of Gag. The 3’ 

primer contained a Bam HI site, linkers that removed the Gag STOP codon, and bases 

that overlapped with the 3’ end of Gag. 2) The PCR product was digested with Kpn I and 

Bam HI and ligated into KpnI/BamHI cut pEGFP-N1. Note that the protein product 

contains a linker of 10 amino acids (NRNGDPPVAT) between the end of Gag and the 

beginning of EGFP. 

This construct allows imaging of intracellular Gag trafficking and localization in live cells. 


Recommended 

Storage: 





REV: 11/08/2015 Page 1 of 2

Contributor: 

Marilyn D. Resh, Ph.D. 

George Pavlakis, M.D., Ph.D. 

References: 

Hermida-Matsumoto, L. and Resh, M.D. (2000), Localization of Human Immunodeficiency 

Virus Type 1 Gag and Env at the Plasma Membrane by Confocal Imaging. J. Virol. 74: 

8670-8679. [Abstract] 

Lindwasser, O.W. and Resh, M.D. (2002) Myristoylation as a target for inhibiting HIV 

assembly: Unsaturated fatty acids block viral budding. Proc. Natl. Acad. Sci. USA. 99: 

13037-13042. [Abstract] 

Perlman, M. and Resh, M.D. (2006), Identification of an intracellular trafficking and 

assembly pathway for HIV-1 Gag. Traffic 7: 731-745. [Abstract] 

Schwartz, S., Campbell, M., Nasioulas, G., Harrison, J., Felber, B.K., Pavlakis, G.N. (1992) 

Mutational inactivation of an inhibitory sequence in human immunodeficiency virus type 1 

results in Rev-independent gag expression. J. Virol. 66: 7176-7182. [Abstract] 

Pavlakis et al. United States Patent 5,965,726; Method of eliminating inhibitory/instability 

regions of mRNA. 

Pavlakis et al. United States Patent 5,972,596; Nucleic acid constructs containing HIV 

genes with mutated inhibitory/instability regions and methods of using same. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pGag-EGFP 

(Cat#11468) from Dr. Marilyn Resh." Also include the reference cited above in any 

publications. 


Category C Reagent (Cat #11468) must contact Dr. George Pavlakis, Human 

Retrovirus Section, Vaccine Branch, Center for Cancer Research, National Cancer 

Institute, NCI-Frederick, PO Box B, Bldg. 535, Frederick MD 21702-1201, Tel: 

(301)846-1474; Fax:(301)846-6368; Email: gp22p@nih.gov, and Andrew D. 

Maslow, Director, Office of Industrial Affairs, Memorial Sloan-Kettering Cancer 

Center, 1275 York Avenue, Box 524, New York, NY 10021, Email: 

maslowa@mskcc.org, Phone: 212-639-6181, Fax: 212-717-3439, before the 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pPA-GFP-GAG 





Description: 

An expression vector that produces fluorescently (GFP) tagged HIV-1 Gag. Used to 

label HIV for viral tracking and imaging. Photo activation allows for discrimination 

between viral signal and high fluorescent background. 

Cloning Vector: Clontech ppaGFP, C1 

Special 


Sequence 

Recommended 

Storage: 



References: 

Gomez CY, Hope TJ. Mobility of human immunodeficiency virus type 1 Pr55Gag in 

living cells. J Virol. 2006 Sep;80(17):8796-806. PubMed PMID: 16912326; PubMed 

Central PMCID: PMC1563866. 

NOTE: 

Acknowledgment for publications should read "pPAGFP-Gag cat#12483 was obtained 






REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pGag-EGFP 



Release 

Category: 

C 

Provided: 3 μL purified plasmid DNA at 2.3 μg/μL (in 10mM Tris, 1mM EDTA, pH 7.4). Host: DH5a 

Description: 

The insert consists of Rev-independent HIV-1 Gag coding sequences [pCMV55M1-10 

obtained from Dr. George Pavlakis, NCI (Schwartz et al (1992) J. Virol. 66: 7176-7182] 

fused in frame to EGFP. The clone was constructed as follows: 1) Rev independent Gag 

was amplified by PCR. The 5’ primer contained a Kpn I site and overlapped with the start 

of the coding region of Gag. The 3’ primer contained a Bam HI site, linkers that removed 

the Gag STOP codon, and bases that overlapped with the 3’ end of Gag. 2) The PCR 

product was digested with Kpn I and Bam HI and ligated into KpnI/BamHI cut pEGFP-N1 

(Clontech). Note that the protein product contains a linker of 10 amino acids 

(NRNGDPPVAT) between the end of Gag and the beginning of EGFP. 

Click here to see the sequence file. 

Cloning Site: KpnI/BamHI 


pEGFP-NI (Kan R ). The size of the insert is 1530 bp. The size of the vector (including 

insert) is 6274 bp. 

Special 


This cDNA directs Rev-Independent expression of an HIV-1 Gag-EGFP fusion protein, 

thereby allowing imaging of intracellular Gag trafficking and localization in live cells. 

Gag-EGFP forms virus-like particles with an efficiency equivalent to that of Gag. 

Recommended 

Storage: 





REV: 11/08/2015 Page 1 of 2

Contributor: 

Marilyn D. Resh, Ph.D. 

George Pavlakis, M.D., Ph.D. 

References: 

Hermida-Matsumoto, L. and Resh, M.D. (2000), Localization of Human Immunodeficiency 

Virus Type 1 Gag and Env at the Plasma Membrane by Confocal Imaging. J. Virol. 74: 

8670-8679. [Abstract] 

Lindwasser, O.W. and Resh, M.D. (2002) Myristoylation as a target for inhibiting HIV 

assembly: Unsaturated fatty acids block viral budding. Proc. Natl. Acad. Sci. USA. 99: 

13037-13042. [Abstract] 

Perlman, M. and Resh, M.D. (2006), Identification of an intracellular trafficking and 

assembly pathway for HIV-1 Gag. Traffic 7: 731-745. [Abstract] 

Schwartz, S., Campbell, M., Nasioulas, G., Harrison, J., Felber, B.K., Pavlakis, G.N. (1992) 

Mutational inactivation of an inhibitory sequence in human immunodeficiency virus type 1 

results in Rev-independent gag expression. J. Virol. 66: 7176-7182. [Abstract] 

Pavlakis et al. United States Patent 5,965,726; Method of eliminating inhibitory/instability 

regions of mRNA. 

Pavlakis et al. United States Patent 5,972,596; Nucleic acid constructs containing HIV 

genes with mutated inhibitory/instability regions and methods of using same. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pGag-EGFP 

(Cat#11468) from Dr. Marilyn Resh." Also include the reference cited above in any 

publications. 


Category C Reagent (Cat #11468) must contact Dr. George Pavlakis, Human 

Retrovirus Section, Vaccine Branch, Center for Cancer Research, National Cancer 

Institute, NCI-Frederick, PO Box B, Bldg. 535, Frederick MD 21702-1201, Tel: 

(301)846-1474; Fax:(301)846-6368; Email: gp22p@nih.gov, and Andrew D. 

Maslow, Director, Office of Industrial Affairs, Memorial Sloan-Kettering Cancer 

Center, 1275 York Avenue, Box 524, New York, NY 10021, Email: 

maslowa@mskcc.org, Phone: 212-639-6181, Fax: 212-717-3439, before the 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pWISP93-93 




Provided: 1 vial of transformed DH5α bacteria in LB containing 20% glycerol ampicillin-resistant. 

Description: 

A 399 bp insert containing the HIV-1 matrix coding sequence from pNL4-3 was isolated 

by PCR. The first three codons were optimized for bacterial expression, with 

NdeI/BamHI sites and start and stop codons added. The insert was cloned into the 

NdeI/BamHI site of pET16b (NdeI = start). Size of cloning vector + insert is 6102 bp. 

Click here to see the plasmid map, etc. for this reagent. 

Click here to see the sequence file for this reagent. 

Special 


This clone is designed for bacterial expression of the HIV-1 matrix protein. Protein 

expression is induced with IPTG in BL21(DE3) competent bacteria in log growth phase. 

As with all pET systems, expression is driven by a T7 promoter. The host bacteria 

strain contains the T7 RNA polymerase gene under lacUV5 control. 

Recommended 

Storage: 

-80°C. 

Contributor: Dr. Wes Sundquist. 

References: 

Massiah MA, Starich MR, Paschall C, Summers MF, Christensen AM, Sundquist WI. 

Three-dimensional structure of the human immunodeficiency virus type 1 matrix 

protein. J Mol Biol 224:198-223, 1994. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pWISP93-93 

from Dr. Wes Sundquist." Also include the reference cited above in any publications. 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pWISP98-85 




Provided: 1 vial of transformed DH5a bacteria in LB containing 20% glycerol ampicillin-resistant. 

Description: 

A 699 bp insert containing the HIV-1 capsid coding sequence from pNL4-3 was isolated 

by PCR. The first three codons were optimized for bacterial expression, with 

NdeI-BamHI sites and start and stop codons added. The insert was cloned into the 

NdeI-BamHI site of pET11a (Novagen) (NdeI = start). Size of cloning vector + insert is 

6338 bp. 

Click here to see the plasmid map, etc. for this reagent. 

Click here to see the sequence file for this reagent. 

Special 


This clone is designed for bacterial expression of the HIV-1 capsid protein. Protein 

expression is induced with IPTG in BL21(DE3) competent bacteria (Novagen) in log 

growth phase. As with all pET systems, expression is driven by a T7 promoter. The 

host strain contains theT7 RNA polymerase gene under lacUV5 control. 

Recommended 

Storage: 

4°C. 

Contributor: Dr. Wes Sundquist. 

References: 

Yoo S, Myszka DG, Yeh C, McMurray M, Hill CP, Sundquist WI. Molecular recognition in 

the HIV-1 capsid/cyclophilin A complex. J Mol Biol 269:780-795, 1997. (Note: gene 

has been moved into pET11a from pET3 since this publication, but the protocol remains 

the same). 

Sundquist W, et al, Unpublished data. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pWISP98-85 

from Dr. Wes Sundquist." Also include the references cited above in any publications. 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

p55 GAG/GFP 





Description: 

p55 GAG/GFP expresses a full-length gag gene product fused to green fluorescent 

protein (GFP). The T7 promoter is utilized for expression by first infecting cells with 

recombinant vaccinia virus vTF 7-3, then transfecting the plasmid DNA. This clone 

allows fluorescent detection of Gag protein localization on living cells. 

Cloning Site: 

Gag gene: NcoI-BamHI (polylinker sites) 

GFP gene: BamHI-EagI sites 

Cloning Vector: pTM1. Amp resistance. 

Special 


An NcoI - BamHI fragment encompassing the gag gene from HXB2 was derived by PCR 

and inserted into the NcoI and BamHI sites of the pTM1 polylinker. An eGFP cassette 

was derived from plasmid pEFGP-N3 (Clontech) as a BamHI -NotI fragment, and 

ligated into pTM1 containing the gag cassette using the BamHI and Eagl sites. Size of 

cloning vector + insert is 6775bp. 



Recommended 

Storage: 


Contributor: Dr. Paul Spearman. 




REV: 11/08/2015 Page 1 of 2

References: 

Sandefur S, Varthakavi V, Spearman P. The I Domain is Required for Efficient Plasma 

Membrane Binding of Human Immunodeficiency Virus Type 1 Pr55Gag. J Virol 

72:2723-2732, 1998. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: p55 GAG/GFP 

from Dr. Paul Spearman." 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: p01-295 


Lot Number: 1 


Provided: 2.5 μg (25 μl) plasmid DNA in TE buffer. Propagate in DH5α. 

Description: 

The insert was derived from synthetic (human codon-optimized) DNA. HIV clade C 

gag coding region is present. This plasmid lacks a promoter and therefore the insert 

should be subcloned into a suitable vector for expression in mammalian cells. 

Cloning Site: 2.5 µg (25 µl) plasmid DNA in TE buffer. Propagate in DH5α. 


The cloning vector is pCR-Script-Amp (Stratagene), the size of the cloning vector 

including the insert is 4.7 kb. 

Special 


This clone is unique because of human codon optimization. The sequence has been 

mapped. 

Recommended 

Storage: 

4°C 

Contributor: Dr. Stephen Dewhurst. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: p01-295 from 

Dr. Stephen Dewhurst." 




REV: 11/08/2015 Page 1 of 2


reagent must contact the Office of Technology Transfer, University of 

Rochester Medical Center, Tel: 585-273-3743 before the reagent can be 

released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: p01-426 


Lot Number: 1 


Provided: 2.5 µg (25 µl) plasmid DNA in TE buffer. Propagate in DH5α. 

Description: 

The insert was derived from synthetic (human codon-optimized) DNA. SIVmac239 

Gag coding region is present. This plasmid lacks a promoter and therefore the insert 

should be subcloned into a suitable vector for expression in mammalian cells. 


Cloning Site: KpnI/SacI cloning site. The size of the insert is 1.7 kb. 

Cloning Vector: The cloning vector is pUC19, the size of the cloning vector including the insert is 4.7 

kb. 

Special 


This clone is unique because of human codon optimization. The sequence has been 

mapped. 

Recommended 

Storage: 

4°C. 

Contributor: Dr. Stephen Dewhurst. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: p01-426 from 

Dr. Stephen Dewhurst." 


reagent must contact the Office of Technology Transfer, University of 





REV: 11/08/2015 Page 1 of 2


released. 




REV: 11/08/2015 Page 2 of 2

HIV-1 >> NEF

DATA SHEET 

Reagent: 

pNef-ER 




Provided: 1 vial transformed DH5α 

Description: 

A Nef protein (NL4-3) that is readily expressed in T-cell lines such as Sup-T1 (Catalog 

#100) and whose function is inducibly activated. It is composed of a fusion between 

full-length Nef and the estrogen receptor hormone-binding domain (Nef-ER). The Nef-ER 

is kept in an inactive state due to steric hindrance, and addition of the 

membrane-permeable drug 4-hydroxytamoxifen (4-HT), which binds to the ER domain, 

leads to inducible activation of Nef-ER within cells. 

Click here for the sequence file. 

Click here for Research Support Documentation 

Cloning Site: Not I – Not I. The size of the insert is 2308 bp (Nef-ER-IRES-puro). 

Cloning Vector: The cloning vector is pEBB. The size of the cloning vector including the insert is 8276 bp. 

Special 


First description of a regulatable Nef. 

Recommended 

Storage: 

-70°C 

Contributor: Drs. Scott Walk, Kodi Ravichandran and David Rekosh. 




REV: 11/08/2015 Page 1 of 2

References: 

Walk SF, Alexander M, Maier B, Hammarskjold M-L, Rekosh DM, Ravichandran KS. 

Design and use of an inducibly activated human immunodeficiency virus type 1 Nef to 

study immune modulation. J Virol 75:834-843, 2001. 

NOTE: 


through the NIH AIDS Reagent Program, NIAID, NIH: pNef-ER from Drs. Scott Walk, 

Kodi Ravichandran and David Rekosh." Also include the reference cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

plconsnefSN 




Provided: 1 vial transformed STBL-2 cells. 

Description: 

Confers ampicillin resistance in bacteria and G418 resistance in animal cells. The 

artificially generated, full-length consensus nef gene was derived from analysis of 54 

HIV-1 patient isolates using overlapping PCR amplification, and cloned into the EcoRI 

site of the MuLV vector LXSN. 

Plasmid Map 

Sequence 

Cloning Site: EcoRI. 

Cloning Vector: MuLV vector LXSN. 

Special 


Confers ampicillin resistance in bacteria and G418 resistance in animal cells. For 

expression of Nef in mammalian cells following packaging using a MuLV packaging cell 

line. The artificially generated, full-length consensus nef gene (639 bp) was derived 

from analysis of 54 patient isolates of HIV-1 using overlapping PCR amplification. The 

nef gene is transducible by retrovirus mediated gene transfer. 

Recommended 

Storage: 

-80°C 

Contributor: 

Dr. Ron Swanstrom, University of North Carolina at Chapel Hill, Lineberger Cancer 

Center. 




REV: 11/08/2015 Page 1 of 2

References: Anderson S, Shugars DC, Swanstrom R, Garcia JV. J Virol 67:4923-4931, 1993. 

Miller AD, Rosman GJ. BioTechniques 7:980-990, 1989. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: plconsnefSN 

from Dr. Ron Swanstrom." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

plconsnefSN 





Description: Full-length consensus nef gene derived from the analysis of 54 HIV-1 patient isolates. 

Cloning Site: EcoRI. 


MuLV vector LXSN. Confers ampicillin resistance in bacteria and G418 resistance in 

animal cells. 

Special 


Full-length consensus nef gene (639 bp) was derived using overlapping PCR 

amplification. The nef gene is transducible by retrovirus mediated gene transfer. For 

expression of Nef in mammalian cells following packaging using a MuLV packaging cell 

line. 


Recommended 

Storage: 


Contributor: 


Center. 




REV: 11/08/2015 Page 1 of 2

References: 

Anderson S, Shugars DC, Swanstrom R, Garcia JV. Nef from primary isolates of 

human immunodeficiency virus type 1 suppresses surface CD4 expression in human 

and mouse T cells. Journal of Virology. 1993;67(8):4923-4931. Abstract 

Miller, A. D., & Rosman, G. J. (1989). Improved Retroviral Vectors for Gene Transfer 

and Expression. BioTechniques, 7(9), 980–990. Abstract 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: plconsnefSN 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1 SF2 Nef 




Provided: 5µL purified plasmid DNA at 1µg/µL. 

Description: 

The insert was cloned by PCR using primers containing EcoR I sites. The resulting 

product was cloned as an EcoR I fragment into pcDNA3.1. Ampicillin resistant. 

Cloning Site: 

The insert is cloned into EcoRI site of pcDNA3.1 and is 661bp The size of the insert is 

661bp. 

Cloning Vector: The size of the cloning vector including insert is 6089bp. 

Special 


Expresses high levels of HIV-1 SF2 Nef 

Host Strain: Grow in DH5α. 

Recommended 

Storage: 

-20°C 

Contributor: Drs. J. Victor Garcia and John Foster 

References: 

Alexa Raney, Lillian S. Kuo, Laura L. Baugh, John L. Foster, and J. Victor Garcia. 

Reconstitution and Molecular Analysis of an Active Human Immunodeficiency Virus Type 

1 Nef/p21-Activated Kinase 2 Complex. J. Virol. 2005 79: 12732-12741. [Abstract] 

[Full Text] [PDF] 

Eduardo O'Neill, Lillian S. Kuo, John F. Krisko, Diana R. Tomchick, J. Victor Garcia, and 

John L. Foster. Dynamic Evolution of the Human Immunodeficiency Virus Type 1 

Pathogenic Factor, Nef J. Virol. 2006 80: 1311-1320. [Abstract] [Full Text] [PDF] 

[Supplemental material] 




REV: 11/08/2015 Page 1 of 2


NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcDNA3.1SF2Nef 

(Cat#11431) from Dr. J. Victor Garcia." Also include the reference cited above in any 

publications. 


reagent must contact Dr. J. Victor Garcia at email address 

victor.garcia@utsouthwestern.edu and specify in the email the name of the 

reagent and a description of the intended use of the reagent. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

p96ZM651nef-opt 





Description: 

This expression vector produces Nef protein which is a codon-optimized form of the 

96ZM651.8 clone (Cat# 6173). 





Special 


Derived from the 96ZM651.8 clone (Cat# 6173). For this construct, the full-length nef 

gene was artificially reconstructed by substituting original viral codons with those of 

highly expressed human genes (Hass J, et al. Curr Biol 6:315-324, 1996). The nef 

amino acid sequence is unchanged. Nine base pairs of wildtype viral sequence preceding 

the initiation codon were retained in this synthetic gene. 

The size of the translated nef protein is 206 aa. 










REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 



References: 





NOTE: 



p96ZM651nef-opt from Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn." Also include 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pT7consnefhis6 




Provided: 5 &#956g of DNA at 0.6 &#956g/&#956l in TE buffer 

Description: 

Confers ampicillin resistance. The artificially generated consensus nef gene was 

derived from analysis of 54 HIV-1 patient isolates using overlapping PCR amplification. 

The full-length nef gene was cloned into the NdeI-PstI site of pT7-7 (from Stan Tabor), 

and is expressed using a T7 RNA polymerase promoter. 

Plasmid Map 

Sequence 

Cloning Site: NdeI to PstI. Insert is 0.6 kb. Total plasmid size is 3.5 kb. 

Cloning Vector: pT7-7 (from Stan Tabor). Vector is 2.9 kb. 

Special 


The full-length nef gene is expressed using a T7 RNA polymerase promoter. The Nef 

protein produced contains six additional C-terminal histidines to allow affinity 

purification using an NTA column. 

Contributor: 


Center. 

References: 

Shugars DC, Smith MS, Glueck DH, Nantermet PV, Seillier-Moiseiwitsch F, Swanstrom J 

R. J Virol 67:4639-4650, 1993. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the AIDS Reagent Program, Division of AIDS, NIAID, NIH: pT7consnefhis6 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1SF2NefF195I 




Provided: 5.0 µl, 1.0 µg/µl. 

Description: 


product was cloned as an EcoR I fragment into pcDNA3.1. Mutations were introduced by 

overlapping PCR mutagenesis and the resulting products cloned as Xho I/ BSPE I 

fragments. Ampicillin resistant. 

Cloning Site: The insert is cloned into EcoRI site of pcDNA3.1 The size of the insert is 661bp. 


Special 


Expresses a Pak-2 defective Nef. 


Recommended 

Storage: 

-20°C 


References: 



1 Nef/p21-Activated Kinase 2 Complex. J. Virol. 2005 79: 12732-12741. [Abstract] [Full 

Text] [PDF] 




REV: 11/08/2015 Page 1 of 2

NOTE: 



pcDNA3.1SF2NefF195I (Cat#11429) from Dr. J. Victor Garcia." Also include the 









REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1SF2NefF195I 





Description: 


product was cloned as an EcoR I fragment into pcDNA3.1. Mutations were introduced by 

overlapping PCR mutagenesis and the resulting products cloned as Xho I/ BSPE I 

fragments. Ampicillin resistant. 

Cloning Site: The insert is cloned into EcoRI site of pcDNA3.1 The size of the insert is 661bp. 


Special 




Recommended 

Storage: 

-20°C 


References: 



1 Nef/p21-Activated Kinase 2 Complex. J. Virol. 2005 79: 12732-12741. [Abstract] [Full 

Text] [PDF] 




REV: 11/08/2015 Page 1 of 2

NOTE: 



pcDNA3.1SF2NefF195I (Cat#11429) from Dr. J. Victor Garcia." Also include the 









REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1SF2NefF195R 





Description: 


product was cloned as an EcoR I fragment into pcDNA3.1. Ampicillin resistant. 

Cloning Site: The insert is cloned into the EcoR I site of pcDNA3.1 The size of the insert is 661bp. 


Special 




Recommended 

Storage: 

-20°C 

Contributor: Drs. J. Victor Garcia and Eduardo O’Neill 

References: 

Eduardo O'Neill, Lillian S. Kuo, John F. Krisko, Diana R. Tomchick, J. Victor Garcia, and 

John L. Foster. Dynamic Evolution of the Human Immunodeficiency Virus Type 1 

Pathogenic Factor, Nef J. Virol. 2006 80: 1311-1320. [Abstract] [Full Text] [PDF] 





REV: 11/08/2015 Page 1 of 2

NOTE: 



pcDNA3.1SF2NefF195R (Cat#11430) from Dr. J. Victor Garcia." Also include the 









REV: 11/08/2015 Page 2 of 2

HIV-1 >> POL

DATA SHEET 

Reagent: 

pINSD.His 



Release 

Category: 

C 

Provided: 5 µL purified plasmid DNA (1 µg/µL). 

Description: 

Contains the NL4-3 (Catalog #114) integrase coding sequence from pINSD (Catalog 

#2820) cloned into pET15B. 

Special 


HIV-1 integrase with an amino-terminal polyhistidine tag is expressed upon 

transformation of pINSD.His into E. coli BL21(DE3) and IPTG induction. The expressed 

integrase may be purified by Ni-affinity chromatography. The polyhistidine tag may be 

subsequently removed by cleavage with thrombin. The integrase is functional in in vitro 

integration assays. E. coli strain BL21(DE3) (Novagen; ph: 1-800-526-7319, Cat. 

#69367-1) must be used for integrase expression, but the plasmid may be unstable in 

this host. Backup stocks should be maintained in HB101. 

NL4-3 Integrase Sequence (pINSD-IN insert) 

pET15B vector 

Recommended 

Storage: 

-80°C. 

Contributor: Dr. Robert Craigie. 




REV: 11/08/2015 Page 1 of 2

References: 

Bushman FD, Engelman A, Palmer I, Wingfield P, Craigie R. Domains of the integrase 

protein of human immunodeficiency virus type 1 responsible for polynucleotidyl transfer 

and zinc binding. Proc Natl Acad Sci USA 90:3428-3432. 

Craigie R, Hickman AB, Engelman A. Integrase In: HIV Volume 2: A Practical Approach. 

J. Karn (Ed.), Oxford Univ Press, pp. 53-71, 1995. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH from Dr. Robert 

Craigie: pINSD.His." Also include the references cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pLJS10 



Release 

Category: 

B 

Provided: 1 vial of ampicillin-resistant transformed bacteria 

Description: 

The HXB2 integrase coding sequence from p22K56 was amplified by PCR, introducing 

ATG and GGG codons 5' to the integrase region. KpnI and NcoI sites were inserted 5' to 

the integrase fragment, and a BamHI site was inserted at the 3' end. The resultant DNA 

fragment was inserted into the KpnI-BamHI site of pUC18. The NcoI-BamHI fragment was 

then isolated and inserted into pET-8C. 

Plasmid Map 

Cloning Site: BamHI 

Cloning Vector: pET-8C 

Special 


HIV-1 integrase can be produced by transformation of this plasmid into E. coli 

BL21(DE3)pLysS bacteria and IPTG induction. Active integrase can be purified from this 

material. The integrase contains additional N-terminal Met and Gly amino acids. Under 

optimal conditions, 20% of the total protein produced is integrase. Store protein at -80°C; 

protein activity is stable for 1 year when stored at -80°C in 50 mM Tris (pH 7.5), 1 mM 

DTT, 50 mM NaCl, 10% glycerol. The protein is unstable after 1 week of storage at 4°C. 

Expressed integrase should be solubilized in 50 mM Tris (pH 7.5), 1 mM DTT, 50 mM 

NaCl, 10 mM MnCl2 for use in assays. 

Recommended 

Storage: 

-80°C. 




REV: 11/08/2015 Page 1 of 2

Contributor: Dr. James M. Groarke, Dr. Joseph V. Hughes, and Dr. Frank J. Dutko. 

References: 

Hong T, Murphy E, Groarke J, Drilca K. Human immunodeficiency virus type 1 DNA 

integration: fine structure target analysis using synthetic oligonucleotides. J Virol 

67:1127-1131, 1993. 

Sioud M, Drilca K. Prevention of human immunodeficiency virus type 1 integrase 

expression in Escherichia coli by a ribozyme. Proc Natl Acad Sci USA 88:7303-7307, 

1991. 

NOTE: 


through the NIH AIDS Reagent Program,Division of AIDS, NIAID, NIH: pLJS10 from Drs. 

JM Groarke, JV Hughes, and FJ Dutko." Also include the references cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pPolo 




Provided: 

1 ml (6.8 x 10 9 ) of ampicillin-resistant transformed HB101 bacteria. Cells are 

suspended in LB medium supplemented with 50 µg/ml ampicillin and 15% glycerol. 

Description: 

Derived from BH10. Contains a 4978 bp BglII insert (nt 2093-7071) cloned into 

pEXc13, a pBR322 derivative. Pol ORF extends from nt 2093-5126. 

Special 


Contains a BglII insert of 4978 bp from HIV-1BH10, seq. 2093-7071 cloned into pExc13 

(derived from pBR322). Pol ORF extends from 2093-5126. Contains ampr marker. Pol 

sequences are induced following removal of tryptophan from growing cultures. Induced 

proteins are detected optimally 2-3 hours after induction (37°C). Produces protease 

and reverse transcriptase. 

Plasmid Map 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Bruce Korant. 

References: 

Danley DE, Geoghegan KF, Scheld KG, Lee S, Merson JR, Hawrylik SJ, Rickett GA, 

Ammirati MJ, Hobart PM. Crystallizable HIV-1 protease derived from expression of the 

viral pol gene in Escherichia coli. Biochem Biophys Res Commun 165:1043-1050, 

1989. 




REV: 11/08/2015 Page 1 of 2

Ivanoff LA, Towatari T, Ray J, Korant BD, Petteway SR Jr. Expression and site-specific 

mutagenesis of the poliovirus 3C protease in Escherichia coli. Proc Natl Acad Sci USA 

83:5392-5396, 1986. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pPolo from Dr. 

Bruce Korant." Also include the references cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pINSD 


Lot Number: 1 


Provided: 1 vial ampicillin-resistant transformed HB101 bacteria. 

Description: 

Bases 4230-5093 of NL4-3 (Catalog #114) were modified to contain an NdeI site and 

initiator ATG at the 5' end and a BamHI site and TAG termination codon at the 3' end, 

then cloned into pET-2C. pINSD has three base changes from NL4-3 that do not affect 

the IN coding region: A4313-G, A4673-G, and A4676-G. 

Click here to see the plasmid map for this reagent. 

Sequence. 

Special 


HIV-1 integrase is expressed upon transformation of pINSD into E. coli BL21(DE3) and 

IPTG induction. The integrase is functional in in vitro integration assays. E. coli strain 

BL21(DE3) (Novagen; ph: 1-800-526-7319, Cat. #69367-1) must be used for 

expression of integrase, but the plasmid may be unstable in this host. Backup stocks 

should be maintained in HB101. 

Recommended 

Storage: 

-90°C. 

Contributor: Dr. Alan Engelman and Dr. Robert Craigie. 

References: 

Engelman A, Craigie R. Identification of conserved amino acid residues critical for 

human immunodeficiency virus type 1 integrase function in vitro. J Virol 66:6361-6369, 

1992. 

Craigie R, Hickman AB, Engelman A. Integrase In: HIV Volume 2: A Practical Approach. 

J. Karn (Ed.), Oxford Univ Press, pp. 53-71, 1995. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH from Dr. Alan 

Engelman and Dr. Robert Craigie: pINSD." Also include the reference cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

p96ZM651pol-opt 




Provided: 20 μg purified plasmid DNA precipitated in 1/10 volume of 3M sodium acetate and 3 

volumes of ethanol, total volume is 200 μl. 

Description: 

This expression vector produces RT and IN from pol which is a codon-optimized form of 




The cloning vector is pcDNA3.1(-) (Invitrogen). This clone is ampicillin and neomycin 

resistant. 

Special 


Derived from the 96ZM651.8 clone (Cat# 6173). For this construct, the RT and IN 

fusion (no PR) gene was artificially reconstructed by substituting original viral codons 

with those of highly expressed human genes (Hass J, et al. Curr Biol 6:315-324, 1996). 

The pol amino acid sequence is unchanged. A Kozak (GCCGCC) sequence and a 

methionine codon (ATG) were added immediately prior to the first codon of the RT. 

The size of the translated pol protein is 849 aa. 









REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-80°C. 


References: 





NOTE: 



p96ZM651pol-opt from Drs. Yingying Li, Feng Gao, and Beatrice H. Hahn." Also include 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pINSD.His.Sol 



Release 

Category: 

C 

Provided: 1 vial ampicillin-resistant transformed HB101 bacteria. 

Description: 

Contains the HIV-1NL4-3 integrase coding sequence from pINSD (Catalog #2820) cloned into pET15B. 

Modified to introduce the amino acid substitutions Phe 185→Lys and Cys 280→Ser. 

Special 


HIV-1 integrase with an amino-terminal polyhistidine tag and F185K and C280S amino 

acid substitutions is expressed upon transformation of pINSD.His.Sol into E. coli 

BL21(DE3) and IPTG induction. The two amino acid substitutions greatly improve the 

solubility of HIV-1 integrase without affecting in vitro activities. The expressed integrase 

may be purified by Ni-affinity chromatography with a high yield (5-10 mg/liter of culture) 

from the soluble fraction of E. coli lysates. The polyhistidine tag may be subsequently 

removed by cleavage with thrombin. The integrase is functional in in vitro integration 

assays. E. coli strain BL21(DE3) (Novagen; Tel: 1-800-526-7319) must be used for 

integrase expression, but the plasmid may be unstable in this host. Backup stocks should 

be maintained in HB101. 

Recommended 

Storage: 

-80°C. 

Contributor: Dr. Robert Craigie. 

References: 

Jenkins TM, Engelman A, Ghirlando R, Craigie R. A soluble active mutant of HIV-1 

integrase: involvement of both the core and carboxyl-terminal domains in 

multimerization. J Biol Chem 271:7712-7718, 1996. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pINSD.His.Sol 

from Dr. Robert Craigie." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

HIV-1 >> REV

DATA SHEET 

Reagent: 

pCV1 




Provided: 1 ml (5 x 10 8 ) of tetracycline-resistant transformed bacteria. 

Description: 

Derived from pCV, a 7.0 kb mammalian expression vector containing hybrid 

regulatory sequences. Contains 1.5 kb of pBR322 sequences adjacent to cDNA 

sequences. The 1.8 kb insert encodes both Tat and Rev. 

Plasmid Map 

Cloning Site: PstI. 

Special 


Tetracycline resistant. PstI cut gives two fragments of 3.3 kb and 7.0 kb. 3.3 kb = 

1.8 + 1.5 kb. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Flossie Wong-Staal. 

References: 

Arya SK., Guo C, Josephs SF, Wong-Staal, F. Trans-activator gene of human 

T-lymphotropic virus type III (HTLV-III). Science 229:69-73, 1985. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pCV-1 from 

Dr. Flossie Wong-Staal." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcRev 




Provided: 5μg purified plasmid DNA (0.7μg/μl) 

Description: 

Derived from pBC12/CMV/IL-2 (cat#11416) by cleavage with Hind III and Xho I followed 

by insertion of a Rev cDNA from the HXB3 strain of HIV-1 IIIB. 

Cloning Site: 

The insert is cloned into HIND III (5′) to Xho I (3′) sites of pBC12/CMV. Insert size is 

170bp. The size of the cloning vector plus insert is 3959bp. 

Cloning Vector: pBC12/CMV (ampicillin resistant). 

Special 


Expresses HIV-1 Rev. 

Recommended 

Storage: 

-20°C 


References: 

Malim MH, Hauber J, Fenrick R, Cullen BR. Immunodeficiency virus rev trans-activator 

modulates the expression of the viral regulatory genes. Nature. 335(6186):181-3, 

1988. [Abstract] [PDF] 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcRev 

(Cat#11415) from Dr. Bryan R. Cullen." Also include the reference cited above in any 

publications. 


must contact the donor, Dr. Bryan R. Cullen, at the Department of Molecular 








REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pCMV-rev (pRev-1) 




Provided: 5 µg purified plasmid DNA (0.934 µg/µL) in TE. 

Description: 

A 722 bp fragment from pCV-1 containing HIV-1 rev cDNA was obtained by Bsu36I 

digestion. This fragment was blunt end ligated into the BamHI site of pCMV to 

generate the 4972 bp plasmid pCMV-rev. This clone has ampicillin resistance. 

Cloning Site: BamHI 

Cloning Vector: pCMV 

Special 


Expresses rev protein upon transfection into mammalian cells. Expression is driven by 

the simian CMV immediate early promoter. The rev cDNA is identical to the rev 

sequences cloned into plasmid pSV-rev (Catalog #1444), and is positioned between 

the CMV promoter and the rabbit ß-globin splice and poly A signals. 

Source of Pro Virus: pCV-1 (cDNA clone, Catalog #303). 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Marie-Louise Hammarskjöld and Dr. David Rekosh. 

References: 

Lewis N, Williams J, Rekosh D, Hammarskjöld M-L. Identification of a cis-acting 

element in human immunodeficiency virus type 2 (HIV-2) that is responsive to the 

HIV-1 rev and human T-cell leukemia virus types I and II rex proteins. J Virol 

64:1690-1697, 1990. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pCMV-rev from 

Dr. Marie-Louise Hammarskjöld and Dr. David Rekosh." Also include the reference 

cited above in any publications. 

Corporate requests should be directed in writing to Dr. David Rekosh or Dr. 

Marie-Louise Hammarskjöld at the University of Virginia, Department of 

Microbiology, HSC Box 441, Charlottesville, VA 22908. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

HIV-1 HXB2 Rev Expression Vector (pHGB1-Rev HXB2) 





Description: This expression vector produces HXB2 Rev. 

Cloning Site: 5’: NcoI 3’: XhoI 

Cloning Vector: Vector (total): 5958 bp Insert: 358 bp 

Special 


Rev from HIV-1 HXB2 fused to an N-terminal GB1 tag for solubility and a hexahistidine 

tag for purification using Ni-NTA chromatography. Rev can be cleaved from these tags 

by tobacco etch virus, resulting in a Rev with an extra Gly-Ala dipeptide at the 

N-terminus. 

It is suggested to use E. coli BL21(DE3) for protein production. 

Recommended 

Storage: 


Contributor: Dr. Alan Frankel 

References: 

Daugherty MD, D'Orso I, Frankel AD. A solution to limited genomic capacity:using 

adaptable binding surfaces to assemble the functional HIV Rev oligomer on RNA. Mol 

Cell. 2008 Sep 26;31(6):824-34. doi: 10.1016/j.molcel.2008.07.016. PMID: 18922466; 

PMCID: PMC2651398. 

Daugherty MD, Booth DS, Jayaraman B, Cheng Y, Frankel AD. HIV Rev response 

element (RRE) directs assembly of the Rev homooligomer into discrete asymmetric 

complexes. Proc Natl Acad Sci U S A. 2010 Jul 13;107(28):12481-6. doi: 




REV: 11/08/2015 Page 1 of 2


10.1073/pnas.1007022107. Epub 2010 Jun 28. PMID: 20616058; PMCID: 

PMC2906596. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HIV-1 HXB2 Rev 

Expression Vector (pHGB1-Rev HXB2) from Dr Alan Frankel." Also include the reference 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

HIV-1 HXB2 Rev Expression Vector (pHGB1-Rev HXB2) 





Description: This expression vector produces HXB2 Rev. 

Cloning Site: 5’: NcoI 3’: XhoI 

Cloning Vector: Vector (total): 5958 bp Insert: 358 bp 

Special 


Rev from HIV-1 HXB2 fused to an N-terminal GB1 tag for solubility and a hexahistidine 

tag for purification using Ni-NTA chromatography. Rev can be cleaved from these tags 

by tobacco etch virus, resulting in a Rev with an extra Gly-Ala dipeptide at the 

N-terminus. 

It is suggested to use E. coli BL21(DE3) for protein production. 

Recommended 

Storage: 


Contributor: Dr. Alan Frankel 

References: 

Daugherty MD, D'Orso I, Frankel AD. A solution to limited genomic capacity:using 

adaptable binding surfaces to assemble the functional HIV Rev oligomer on RNA. Mol 

Cell. 2008 Sep 26;31(6):824-34. doi: 10.1016/j.molcel.2008.07.016. PMID: 18922466; 

PMCID: PMC2651398. 

Daugherty MD, Booth DS, Jayaraman B, Cheng Y, Frankel AD. HIV Rev response 

element (RRE) directs assembly of the Rev homooligomer into discrete asymmetric 





REV: 11/08/2015 Page 1 of 2


10.1073/pnas.1007022107. Epub 2010 Jun 28. PMID: 20616058; PMCID: 

PMC2906596. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HIV-1 HXB2 Rev 

Expression Vector (pHGB1-Rev HXB2) from Dr Alan Frankel." Also include the reference 





REV: 11/08/2015 Page 2 of 2

HIV-1 >> TAT

DATA SHEET 

Reagent: 

pGST-Tat 1 86R 



Release 

Category: 

B 


Description: 

See attached Table. These constructs contain full length wild type or mutant HIV-1 

(HXB2) or HIV-2 (ROD) tat genes. The clones contain a thrombin proteolytic site between 

the Schistosoma japonicum glutathione S-transferase (GST) sequence and the tat insert. 

Tat is expressed as a GST fusion protein. Constructs cloned into the pGEX2TK vector 

contain the phosphorylation site for the catalytic subunit of camp-dependent heart 

muscle kinase. GST-Tat fusions in this vector can thus be labeled directly in vitro to high 

specific activities with (g32P)ATP and commercially available kinase (Sigma). 

Table. HIV Tat Expression Vectors 

Cloning Vector: pGEX2T or pGEX2TK (Pharmacia). 

Special 


The HIV-1 constructs encode Tat in full length (86 aa), first exon (72 aa), activation 

domain (first 48 aa), or mutated forms. HIV-2 Tat is encoded as full length (130 aa), one 

exon (99 aa), or mutated protein. The GST-Tat fusion proteins can be easily purified 

from E. coli lysates using a single-step procedure under non-denaturing conditions (see 

protocol, page 8). Tat can be cleaved from the fusion proteins and purified by thrombin 

proteolytic digestion. 

Protocol: Preparation and Purification of HIV-1/2 Tat 

Recommended 

Storage: 

-80°C. 

Contributor: Dr. Andrew Rice. 




REV: 11/08/2015 Page 1 of 2

References: 

Herrmann CH, Rice AP. Lentivirus Tat proteins specifically associate with a cellular protein 

kinase, TAK, that hyperphosphates the carboxyl-terminal domain of the large subunit of 

RNA polymerase II: candidate for a Tat cofactor. J Virol 69:1612-1620, 1995. 

Rhim H, Echetebu CO, Herrmann CH, Rice AP. Wild type and mutant HIV-1 and HIV-2 Tat 

proteins expressed in Escherechia coli as fusions with glutathione S-transferase. J 

Acquired Immune Defic Syndr 7:1116-1121, 1994. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: (specify reagent) 

from Dr. Andrew Rice." Also include the references cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-80°C. 





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References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA3.1+/Tat101-flag(PEV280) 



Release 

Category: 

C 


Description: 

The plasmid expresses HIV-1 tat protein, a 101 amino acid protein, under the control of 

the CMV promoter. 


The cloning vector is pcDNA 3.1(+). The size of the vector containing the insert is 

approximately 5.7 kb and the size of the insert is 2.3 Kb. pcDNA3.1(+) is ampicillin 

resistant. 

Special 


A PCR-amplified cDNA fragment corresponding to Tat101 of the HIV-1 Tat gene was 

amplified with the use of the HIV-1 cDNA clone pCV1 as a template and primers 

containing an additional HindIII site for cloning. The amplified fragment was cloned into 

the unique HindIII site of pcDNA3.1(+). Because pCV1 cDNA encodes a truncated 86– 

amino acid form of Tat, a mutation (TAG to TCG, changing codon 86 from a stop to a 

serine) was introduced in the primer used in the PCR reaction to reopen the Tat reading 

frame to its full length. 


Recommended 

Storage: 


Contributor: Dr. Eric Verdin 




REV: 11/08/2015 Page 1 of 2

References: 

Ott M, Emiliani S, Van Lint C, Herbein G, Lovett J, Chirmule N, McCloskey T, Pahwa S and 

Verdin E. Immune hyperactivation of HIV-1-infected T cells mediated by Tat and the 

CD28 pathway. Science 275:1481-148, 1997. 

NOTE: 



pcDNA3.1+/tat101-flag from Dr Eric Verdin." Also include the reference cited above in 



must contact Joan Bruland at jbruland@gladstone.ucsf.edu and specify in the 

email the name of the reagent and a description of the intended use of the 

reagent. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pC-Tat.BL43.CC 



Release 

Category: 

C 

Provided: 1 vial containing 5µg of purified DNA in TE buffer (0.6 µg/µl) 

Description: 

HIV-1 subtype C Tat mammalian expression vector and its isogenic mutants in the 

cysteine-rich domain (pC-TatBL43.CS, pC-TatBL43.CC and pC-TatBL43.SC). The 

full-length Tat was amplified using RT-PCR from total RNA extracted from co-cultured 

PBMC of a primary Indian clinical sample BL43/02. The Tat expression cassette was 

cloned into the pcDNA3.1+ vector under the control of the CMV promoter. From the wild 

type Tat expression vector (pC-TatBL43.CS) two isogenic Tat mutants in the cysteine-rich 

domain were generated using site-directed mutagenesis. The mutations are located at 

positions 30 and 31 of Tat, respectively. 

HIV-1 subtype C Tat bacterial expression vectors are also available: pE-TatBL43.CS (Cat 

#11791), pE-TatBL43.CC (Cat #11792) and pE-TatBL43.SC (Cat #11793). 

Cloning Site: The 0.3 kb insert was directionally cloned between EcoRI and XbaI sites of the vector. 

Cloning Vector: pcDNA3.1+ (Invitrogen) 

Special 


The cysteine-rich domain regulates several important function of Tat including 

transactivation, monocyte chemotactic activity, apoptosis and others. The wild type Tat 

vector as well as the two isogenic control vectors will be useful in studying biological 

functions of Tat of subtype C origin. 

The GenBank Accession No. FJ765005.1. 

Plasmid Map 




REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-70°C 

Contributor: Dr. Udaykumar Ranga 

References: Ranga U, Shankarappa R, Siddappa NB, Lakshmi R, Nagendran R, Mahalingam M, 

Mahadevan A, Narayana J, Satishchandra P, Shankar SK and Prasad VR. ‘Tat protein of 

Human Immunodeficiency Virus Type-1 subtype C viruses is a defective chemokine’ J. 

Virol, 78 (5), 2586-2590, 2004. 

M. Mishra, S. Vetrivel, N. B. Siddappa, U. Ranga and P. Seth, Clade Specific Neurotoxicity 

of HIV Tat. Significance of the Dicysteine C30C31 Motif, Annals of Neurology, 63, 366 - 

376 (2008) 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pCTatBL43 from 

Dr. Udaykumar Ranga.”Also include the reference cited above in any publications. Contact 

the contributor for more details. 


must contact Dr. Udaykumar Ranga, Jawaharlal Nehru Centre for Advanced 

Scientific Research, Jakkur Post, Bangalore, 560064, India. Tel: (080) 228-2831 

Email:udaykumar@jncasr.ac.in, before the reagent can be released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pC-Tat.BL43.SC 



Release 

Category: 

C 


Description: 













Special 







Plasmid Map 




REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-70°C 





Virol, 78 (5), 2586-2590, 2004. 



376 (2008) 

NOTE: 












REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pE-Tat.BL43.SC 



Release 

Category: 

C 


Description: 

HIV-1 subtype C Tat bacterial expression vector and its isogenic mutants in the 

cysteine-rich domain (pE-TatBL43.CS, pE-TatBL43.CC and pE-TatBL43.SC). The 



cloned into the pET21b vector. From the wild type Tat expression vector 

(pE-TatBL43.CS), two isogenic Tat mutants in the cysteine-rich domain were generated 

using site-directed mutagenesis. Tat from all the vectors is expressed as a fusion protein 

of a HIS-tag at the C-term. 

HIV-1 subtype C Tat mammalian expression vectors are also available: pC-TatBL43.CS 

(Cat #11786), pC-TatBL43.CC (Cat #11785) and pC-TatBL43.SC (Cat #11787). 

Cloning Site: The 0.3 kb insert was directionally cloned between NdeI and XhoI sites of the vector. 

Cloning Vector: pET21b(Novagen) 

Special 


The constructs encode full-length Tat of 101 aa. 


Plasmid Map 

Recommended 

Storage: 

-70°C 




REV: 11/08/2015 Page 1 of 2





Virol, 78 (5), 2586-2590, 2004. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pE-TatBL43.SC 

from Dr. Udaykumar Ranga.” Also include the reference cited above in any publications. 

Contact the contributor for more details. 








REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 1 48D P18IS 



Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 1 86R P18IS 



Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pGST-Tat 1 86R TK 



Release 

Category: 

B 

Provided: 1 mL ampicillin-resistant, transformed BL21 bacteria (glycerol stock). 

Description: 










Special 









Recommended 

Storage: 

-80°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







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DATA SHEET 

Reagent: 

GST-Tat 1 72R C22G 



Release 

Category: 

B 

Provided: Each clone is provided as 1 vial of ampicillin-resistant, transformed BL21 bacteria. 

Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







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DATA SHEET 

Reagent: 

pE-Tat.BL43.SC 



Release 

Category: 

C 

Provided: 

0.5 µg/µL purified plasmid DNA in TE. The plasmid can be propagated in DH5a. The 

plasmids are ampicillin resistant. 

Other available clones: pE-Tat.BL43.CS (Cat #11791) and pE-Tat.BL43.CC (Cat #11792). 

Description: 













Special 




Plasmid Map 




REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-70°C 





Virol, 78 (5), 2586-2590, 2004. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pE-TatBL43.SC 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pC-Tat.BL43.SC 



Release 

Category: 

C 

Provided: 

0.5 µg/µL purified plasmid DNA in TE. This plasmid can be propagated in DH5a on 

ampicillin plates. 

Other available clones: pC-Tat.BL43.CC (Cat #11785) and pC-Tat.BL43.CS (Cat #11786). 

Description: 













Special 







Plasmid Map 




REV: 11/08/2015 Page 1 of 2

Plasmid Map 

Recommended 

Storage: 

-70°C 





Virol, 78 (5), 2586-2590, 2004. 



376 (2008) 

NOTE: 












REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pC-Tat.BL43.CS 



Release 

Category: 

C 

Provided: 1 vial containing 5µg of purified DNA in TE buffer (0.6 µg/µl) 

Description: 













Special 







Plasmid Map 




REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-70°C 





Virol, 78 (5), 2586-2590, 2004. 



376 (2008) 

NOTE: 












REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pE-Tat.BL43.CS 



Release 

Category: 

C 

Provided: 



Other available clones: pE-Tat.BL43.CC (Cat #11792) and pE-Tat.BL43.SC (Cat #11793). 

Description: 













Special 




Plasmid Map 




REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-70°C 





Virol, 78 (5), 2586-2590, 2004. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pE-TatBL43.CS 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pE-Tat.BL43.CC 



Release 

Category: 

C 

Provided: 



Other available clones: pE-Tat.BL43.CS (Cat #11791) and pE-Tat.BL43.SC (Cat #11793). 

Description: 













Special 




Plasmid Map 




REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-70°C 





Virol, 78 (5), 2586-2590, 2004. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pE-TatBL43.CC 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pTatC6H-1 




Provided: 2 ug plasmid DNA per vial (ampicillin/kanamycin marker) 

Description: 

The full-length tat gene is expressed in pDS56, RB, 6X His. Purification of recombinant 

Tat protein (86 aa) can be accomplished using metal chelate chromatography under 

denaturing conditions. 

Purification of rTat Protein 

Special 


A one liter bacterial culture yields up to 5–10 mg of active Tat protein. Following 

renaturation, active Tat can be used for biological and immunological assays. 

Bacterial Host: M15::pDM1.1 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Abhay Patki and Dr. Michael Lederman. 

References: Patki AH, Lederman MM. Cell Immunol 169:40, 1996 

Purvis SF, et al. AIDS Res Hum Retroviruses 11:443, 1995. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH:pTatC6H-1 

from Dr. Abhay Patki and Dr. Michael Lederman." Also include the reference cited 





REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pSV2tat72 




Provided: 1 ml ampicillin-resistant transformed HB101 bacteria. 

Description: 

Produces Tat (residues 1-72) using the SV40 early promoter. Constructed by 

replacing the dhfr gene in pSV2-dhfr with a synthetic gene encoding Tat. 

Plasmid Map 

Sequence 

Cloning Site: HindIII–BglII 

Cloning Vector: pSV2-dhfr (Subramani, S., et al. Mol. Cell. Biol. 1:854, 1981). 

Special 


Contact contributor for additional information. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Alan Frankel. 

References: 

Frankel AD, Pabo CO. Cellular uptake of the tat protein from human 

immunodeficiency virus. Cell 55:1189–1193, 1988. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pSV2tat72 

from Dr. Alan Frankel." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pGEM2Tat Expression System 




Provided: 1 vial transformed BL21 (DE3) Lys S bacteria. 

Description: 

A 295 bp ARV-2B fragment encoding the first exon of tat was joined to a pET3A 

transcription terminator and this construct cloned as a HincII-EcoRV fragment 

downstream of the pGEM2 T7 promoter. The pGEM2 polylinker SmaI site was 

destroyed during the cloning process. pGEM2Tat contains a b-lac marker. 

Plasmid Map 

Special 


Allows production of high levels of Tat after induction with 0.4 mM IPTG at O.D. = 

0.4-0.8. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Richard Gaynor. 

References: 

Rosenberg AH, Nade BN, Chui D-S, Lin S-W, Dunn JJ, Studier FW. Vectors for selective 

expression of cloned DNAs by T7 RNA polymerase. Gene 56:125, 1987. 

Garcia JA, Hardch D, Pearson L, Mitsuyasu R, Gaynor R. Functional domains required 

for tat-induced transcriptional activation of the HIV-1 long terminal repeat. EMBO J 

7:3143-3147, 1988. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pGEM2Tat 

Expression System from Dr. Richard Gaynor." 




REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 1 48D TK 


Lot Number: 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 1 86R D2/36 TK 


Lot Number: 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 1 48D P18IS 


Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 1 48D 


Lot Number: 4 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 2 130R 


Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 1 48D C22G 


Lot Number: 5 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 



Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 1 72R 


Lot Number: 3 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 



Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 1 86R C22G 


Lot Number: 3 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 




Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

HIV-1 >> VIF

DATA SHEET 

Reagent: 

pcDNA-HVif 




Provided: 5 µg purified plasmid DNA in TE, 1 µg/µL. The plasmid was amplified in JM 109 bacteria. 

Description: 

The pcDNA-HVif vector encodes the Vif protein of HIV-1 NL4-3 under the control of the 

CMV promoter. HVif is a partially codon-optimized version of the native vif gene. Codon 

optimization was achieved by changing the first 84 vif codons to conform to the 

reported codon usage of highly expressed human genes. The composition of the insert 

has been confirmed by DNA sequencing. DNA sequence information available: 

Click here to see the sequence document. 

Click here to see the plasmid map document. 

Cloning Site: 

The 268 bp insert was cloned in sense orientation into the EcoRl-PflMl restriction sites of 

plasmid pcDNA-Vif. (See reference Nguyen et al for more details) 

Cloning Vector: Invitrogen pcDNA3.1(-). The size of the cloning vector including the insert is 6084 bp. 

Special 


The plasmid expresses high levels of the HIV-1 Vif protein independently of other viral 

proteins. 

Recommended 

Storage: 

-20°C (or below). 

Contributor: Dr. Stephan Bour and Dr. Klaus Strebel 




REV: 11/08/2015 Page 1 of 2

References: Nguyen, K-L, Llano, M., Akari, H., Miyagi, E., Poeschla, E. M., Strebel, K. and Bour, S. 

Codon optimization of the HIV-1 vpu and vif genes stabilizes their messenger RNA and 

allows for highly efficient Rev-independent expression. Virology 319:163, 2004. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcDNA-HVif from 

Dr. Stephan Bour and Dr. Klaus Strebel." Also include the reference cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

HIV-1 >> VPU

DATA SHEET 

Reagent: 

pcDNA-Vphu 



Release 

Category: 

B 

Provided: 5 µg purified plasmid DNA, 1.3 µg/µL. The plasmid was amplified in JM 109 bacteria. 

Description: 

The pcDNA-Vphu vector encodes the Vpu protein of HIV-1 NL4-3 under the control of the 

CMV promoter. Vphu is a codon-optimized version of the native vpu gene. Codon 

optimization was achieved by changing vpu codons to conform to the reported codon 

usage of highly expressed human genes. In addition, the Vpu initiation codon was 

optimized according to the Kozak context rules. Finally, the internal env initiation codon 

was inactivated. Expression is from the cytomegalovirus early promoter and is 

constitutive. The composition of the insert has been confirmed by DNA sequencing. 


Cloning Site: 

The 294 bp insert was cloned in sense orientation into the Mscl-Acc65l restriction sites of 

plasmid pcDNA-Vpu. (See reference Nguyen et al for more details). 


Special 


The plasmid expresses high levels of the HIV-1 Vpu protein independently of other viral 

proteins. 

Recommended 

Storage: 

-20°C (or below). 





REV: 11/08/2015 Page 1 of 2




NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcDNA-Vphu from 

Dr. Stephan Bour and Dr. Klaus Strebel." Also include the reference cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcDNA-Vphu 





Description: 

The pcDNA-Vphu vector encodes the Vpu protein of HIV-1 NL4-3 under the control of 

the CMV promoter. Vphu is a codon-optimized version of the native vpu gene. The 

plasmid expresses high levels of the HIV-1 Vpu protein independently of other viral 

proteins. 

Cloning Site: 

The 294 bp insert was cloned in sense orientation into the Mscl-Acc65l restriction sites 

of plasmid pcDNA-Vpu. 


Special 


Codon optimization was achieved by changing vpu codons to conform to the reported 

codon usage of highly expressed human genes. In addition, the Vpu initiation codon was 

optimized according to the Kozak context rules. Finally, the internal env initiation codon 

was inactivated. Expression is from the cytomegalovirus early promoter and is 

constitutive. The composition of the insert has been confirmed by DNA sequencing. 


Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2




NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pcDNA-Vphu 

from Dr. Stephan Bour and Dr. Klaus Strebel." Also include the reference cited above in 





REV: 11/08/2015 Page 2 of 2

HIV-1 >> VPR

DATA SHEET 

Reagent: 

pEGFP-Vpr 





Description: 

The plasmid expresses a fusion protein that includes humanized eGFP at the amino 

terminus and Vpr at the carboxy terminus and is packaged into HIV virions. Plasmid can 

be use to generate fluorescent virions for attachment assays. 

Cloning Site: 

pNL4-3 Vpr was inserted into the HindIII and Xbal sites of the Clontech Vector 

pEGFP-C3 The size of the insert is 300 bp. 


The cloning vector is pEGFP-C3. The size of the cloning vector including the insert is 

4989 bp. Neomycin resistance. 

Special 


The Vpr coding region was amplified using PCR, from a full-length pNL4-3 clone. The 5′ 

oligonucleotide contained a HindIII restriction site that allowed fusion of the eGFP open 

reading frame with that of Vpr. The 3′ oligonucleotide contained a stop codon as well as 

an Xbal restriction site. The mammalian coding sequences include humanized GFP 

(eGFP) fused with Vpr under control of a CMV-IE promoter 

Sequence File. 

Recommended 

Storage: 

-80°C 

Contributor: Dr. Warner C. Greene 




REV: 11/08/2015 Page 1 of 2

References: 

Schaeffer, E., Geleziunas, R., and Greene, Warner C. Human Immunodeficiency Virus 

Type I Nef Functions at the Level of Virus Entry by Enhancing Cytoplasmic Delivery of 

Virions. J of Virol. 75(6): 2993-3000, 2001. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pEGFP-Vpr (cat# 

11386) from Dr. Warner C. Greene." Also include the reference cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pGFP-VPR 




Provided: 5 μg at 0.6 mg/mL of purified DNA in TE buffer. 

Description: 

An expression vector that produces green fluorescent protein (GFP) tagged HIV-1 

Vpr. Used to label HIV for viral tracking and imaging. 

Cloning Vector: Clontech pEGFP 

Recommended 

Storage: 



References: 

McDonald D, Vodicka MA, Lucero G, Svitkina TM, Borisy GG, Emerman M, Hope TJ. 

Visualization of the intracellular behavior of HIV in living cells. J Cell Biol. 2002 Nov 

11;159(3):441-52. 

NOTE: 

Acknowledgment for publications should read "pGFP-VPR cat#12478 was obtained 






REV: 11/08/2015 Page 1 of 1

DATA SHEET 

Reagent: 

pmCherry-VPR 





Description: 

An expression vector that produces fluorescently (mCherry) tagged HIV-1 Vpr. Used 

to label HIV for viral tracking and imaging. 

Cloning Site: Cherry into pCR2.1, then digested out with BglII and NheI. 

Cloning Vector: Clontech pCherry 

Recommended 

Storage: 

-20 o C for long term storage. 


References: 

LeBlanc JJ, Perez O, Hope TJ. Probing the Structural States of Human 

Immunodeficiency Virus Type 1 Pr55gag by Using Monoclonal Antibodies . Journal of 

Virology 2008;82(5):2570-2574. 

NOTE: 

Acknowledgment for publications should read "pmCherry-VPR cat#12479 was 

obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH from 

Dr. Thomas J. Hope." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 1 of 1

DATA SHEET 

Reagent: 

pPA-GFP-VPR 





Description: 

An expression vector that produces green fluorescent protein (GFP) tagged HIV-1 

Vpr. Used to label HIV for viral tracking and imaging. Photo activation allows for 

discrimination between viral signal and high fluorescent background. 

Cloning Vector: Clontech ppaGFP, C1 

Special 


Sequence 

Recommended 

Storage: 



References: 

Campbell EM, Perez O, Melar M, Hope TJ. Labeling HIV-1 virions with two fluorescent 

proteins allows identification of virions that have productively entered the target cell. 

Virology. 2007 Apr 10;360(2):286-93. 

NOTE: 

Acknowledgment for publications should read "pPA-GFP-VPR cat#12482 was obtained 






REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pCMV-FLAG.DCAF-1_ISO1 




Provided: 5 µg purified plasmid DNA. 

Description: 

Expression vector for DCAF-1, also known as VprBP. Insert was cloned in fragments 

from a human cDNA library, and was assembled in pcDNA-3 using accession 

NM_014703 as a guideline. 

Cloning Site: 

HindIII-XhoI 

Insert size: 4,558 bp. 

Cloning Vector: pCMV-FLAG 

Special 


Expression vector is for mammalian cells. Protein produced is DCAF-1, otherwise 

known as VprBP. 

Plasmid Sequence 

Recommended 

Storage: 

-20 °C 

Contributor: Dr. Vicente Planelles 

References: 

Angers, S., Li, T., Yi, X., MacCoss, M.J., Moon, R.T., Zheng, N. Molecular architecture 

and assembly of the DDB1-CUL4A ubiquitin ligase machinery. Nature Letters, 443; 

pp. 590-593 (2006). 

DeHart, J.L., Zimmerman, E.S., Ardon, O., Monteiro-Filho, C.M.R., Arganaraz, E.R., 

Planelles, V. HIV-1 Vpr activates the G2 checkpoint through manipulation of the 

ubiquitin proteasome system. Virology Journal (2007). 




REV: 11/08/2015 Page 1 of 2

ubiquitin proteasome system. Virology Journal (2007). 

NOTE: 



pCMV-FLAG.DCAF-1(Cat# 11690) from Dr. Vicente Planelles." Also include the 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pEGFP-Vpr 




Provided: 15 μg purified plasmid DNA at 1 μg/μL. 

Description: The Vpr coding region was amplified using PCR, from a full-length pNL4-3 clone. The 5 ′ 

oligonucleotide contained a HindIII restriction site that allowed fusion of the eGFP open 

reading frame with that of Vpr. The 3′ oligonucleotide contained a stop codon as well as 

an Xbal restriction site. The mammalian coding sequences include humanized GFP (eGFP) 

fused with Vpr under control of a CMV-IE promoter and a neomycin resistance gene 

under the control of an SV40 promoter. 

Sequence File. 

Cloning Site: 

pNL4-3 Vpr was inserted into the HindIII and Xbal sites of the Clontech Vector pEGFP-C3 



The cloning vector is pEGFP-C3. The size of the cloning vector including the insert is 

4989 bp. 

Special 


The plasmid expresses a fusion protein that includes humanized eGFP at the amino 

terminus and Vpr at the carboxy terminus and is packaged into HIV virions. Plasmid can 

be use to generate fluorescent virions for attachment assays. 

Recommended 

Storage: 

-20°C 

Contributor: Dr. Warner C. Greene 




REV: 11/08/2015 Page 1 of 2

References: 

Schaeffer, E., Geleziunas, R., and Greene, Warner C. Human Immunodeficiency Virus 

Type I Nef Functions at the Level of Virus Entry by Enhancing Cytoplasmic Delivery of 

Virions. J of Virol. 75(6): 2993-3000, 2001. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pEGFP-Vpr (cat# 

11386) from Dr. Warner C. Greene." Also include the reference cited above in any 

publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pMM310 



Release 

Category: 

C 

Provided: 5 μL of purified plasmid DNA (1 μg/μL). 

Description: 

The pMM310 construct encodes Escherichia coli β-lactamase (BlaM) fused to the amino 

terminus of HIV-1 Vpr. The BlaM (β-lactamase) gene sequence was amplified by PCR with 

HindIII and BamHI sites in primers. The Vpr coding sequence was amplified from the 

HIV-1 genome with BamHI and AvaI sites in PCR primers. Both PCR products were cut 

with HindIII, BamHI and AvaI and cloned into HindIII-AvaI digested pcDNA3.1/zeo. 

BlaM-vpr is constitutively expressed in most mammalian cells. Expression of BlaM-vpr is 

driven by the CMV promoter. 

Sequence 

Plasmid Map 

Cloning Site: HindIII - AvaI, (5'→3') 

Cloning Vector: pcDNA3.1/zeo 

Special 


Ability to generate HIV virions containing active β-lactamase for use in entry assays. 

When loaded with CCF2, a fluorogenic substrate of β-lactamase, infected target cells can 

be detected by a shift in emitted fluorescence. 

Recommended 

Storage: 

-20°C or below 

Contributor: Dr. Michael Miller (Merck Research Laboratories) 




REV: 11/08/2015 Page 1 of 2

References: 

Tobiume M, Lineberger JE, Lundquist CA, Miller MD, and Aiken C. Nef Does Not Affect the 

Efficiency of Human Immunodeficiency Virus Type 1 Fusion with Target Cells. J. Virology 

77(19); 10645-50, 2003. [Abstract]. 

An in-depth description of a similar system: Cavrois M, De Noronha C, Greene WC. A 

sensitive and specific enzyme-based assay detecting HIV-1 virion fusion in primary T 

lymphocytes. Nat Biotechnol. 2002 Nov;20(11):1151-4. Epub 2002 Sep 30. [Abstract] 

NOTE: 



pMM310(Cat#11444) from Dr. Michael Miller." Also include the reference cited above in 



must contact D. Gabrielle Brouillette, Research Contracts Management, Merck 

Research Laboratories, 770 Sumneytown Pike, West Point, PA 19486 Tel: 

215-652-4374 Fax: 215-652-3143, e-mail: gabriella_brouillette@merck.com 





REV: 11/08/2015 Page 2 of 2

HIV-2 >> Tat

DATA SHEET 

Reagent: 

GST-Tat 2 90D 


Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pGal4Tat2 (K70A) 




Provided: Each clone provided as 1 ml ampicillin-resistant, transformed bacteria. 

Description: 

These constructs contain mutated HIV-2 tat genes. Inserts were constructed using PCR 

and/or recombinant PCR. Construction of the vector containing the Gal4 DNA binding 

domain fused to mutated HIV-2 tat-2 genes is described in Sadowski et al. 

Special 


These plasmids encoding Gal4 DNA binding/Tat-2 fusion proteins are designed for 

transient expression assays in mammalian cells. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Sandra Tong. 

References: 

Pagtakhan AS, Tong-Starksen SE. Interactions between Tat of HIV-2 and transcription 

factor Sp1. Virology 238: 221-30, 1997. 

Sadowski I, Ptashne M. A vector for expressing GAL4 (1-147) fusions in mammalian 

cells. Nucleic Acids Res 17:7539, 1989. 

NOTE: 



from Dr. Sandra Tong." Also include the reference cited above in any publications. 


Clone 

Cat. 

No. 

Lot 

Description 




REV: 11/08/2015 Page 1 of 2

pGal4Tat2 

(K70A) 

3639 

1 

7/15/97 

Substitution mutation of core region (K70→A) in 

full-length tat-2 cDNA. 

pGal4Tat2 

(R81.84A) 

3640 

1 

7/15/97 

Substitution mutation of basic residues (R81.84) 

to alanine in full-length tat-2 cDNA. 

pGal4Tat2 

(26.45) 

3641 

1 

7/15/97 

Two exons; internal deletion of residues 26.45, 

inclusive. 

pGal4Tat2 

(46.130) 

3645 

1 

7/15/97 

Two exons; residues 1.45 deleted. 

pGal4Tat2 

(18.130) 

3646 

1 

7/15/97 


pGal4Tat2 

(1.80) 

3647 

1 

7/15/97 

One exon; residues 81.130 deleted. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pGal4Tat2 (R81-84A) 





Description: 




Special 




Recommended 

Storage: 

-70°C. 


References: 





NOTE: 





Clone 

Cat. 

No. 

Lot 

Description 




REV: 11/08/2015 Page 1 of 2

pGal4Tat2 

(K70A) 

3639 

1 

7/15/97 



pGal4Tat2 

(R81.84A) 

3640 

1 

7/15/97 



pGal4Tat2 

(26.45) 

3641 

1 

7/15/97 


inclusive. 

pGal4Tat2 

(46.130) 

3645 

1 

7/15/97 


pGal4Tat2 

(18.130) 

3646 

1 

7/15/97 


pGal4Tat2 

(1.80) 

3647 

1 

7/15/97 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: pGal4Tat2 (26-45) 





Description: 




Special 




Recommended 

Storage: 

-70°C. 


References: 





NOTE: 





Clone 

Cat. 

No. 

Lot 

Description 




REV: 11/08/2015 Page 1 of 2

pGal4Tat2 

(K70A) 

3639 

1 

7/15/97 



pGal4Tat2 

(R81.84A) 

3640 

1 

7/15/97 



pGal4Tat2 

(26.45) 

3641 

1 

7/15/97 


inclusive. 

pGal4Tat2 

(46.130) 

3645 

1 

7/15/97 


pGal4Tat2 

(18.130) 

3646 

1 

7/15/97 


pGal4Tat2 

(1.80) 

3647 

1 

7/15/97 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 






Description: 




Special 




Recommended 

Storage: 

-70°C. 


References: 





NOTE: 





Clone 

Cat. 

No. 

Lot 

Description 




REV: 11/08/2015 Page 1 of 2

pGal4Tat2 

(K70A) 

3639 

1 

7/15/97 



pGal4Tat2 

(R81.84A) 

3640 

1 

7/15/97 



pGal4Tat2 

(26.45) 

3641 

1 

7/15/97 


inclusive. 

pGal4Tat2 

(46.130) 

3645 

1 

7/15/97 


pGal4Tat2 

(18.130) 

3646 

1 

7/15/97 


pGal4Tat2 

(1.80) 

3647 

1 

7/15/97 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 






Description: 




Special 




Recommended 

Storage: 

-70°C. 


References: 





NOTE: 





Clone 

Cat. 

No. 

Lot 

Description 




REV: 11/08/2015 Page 1 of 2

pGal4Tat2 

(K70A) 

3639 

1 

7/15/97 



pGal4Tat2 

(R81.84A) 

3640 

1 

7/15/97 



pGal4Tat2 

(26.45) 

3641 

1 

7/15/97 


inclusive. 

pGal4Tat2 

(46.130) 

3645 

1 

7/15/97 


pGal4Tat2 

(18.130) 

3646 

1 

7/15/97 


pGal4Tat2 

(1.80) 

3647 

1 

7/15/97 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 






Description: 




Special 




Recommended 

Storage: 

-70°C. 


References: 





NOTE: 





Clone 

Cat. 

No. 

Lot 

Description 




REV: 11/08/2015 Page 1 of 2

pGal4Tat2 

(K70A) 

3639 

1 

7/15/97 



pGal4Tat2 

(R81.84A) 

3640 

1 

7/15/97 



pGal4Tat2 

(26.45) 

3641 

1 

7/15/97 


inclusive. 

pGal4Tat2 

(46.130) 

3645 

1 

7/15/97 


pGal4Tat2 

(18.130) 

3646 

1 

7/15/97 


pGal4Tat2 

(1.80) 

3647 

1 

7/15/97 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat2 (R81-84A) 


Lot Number: 1 

Release 

Category: 

A 


Description: 

These constructs contain mutated HIV-2 tat genes. A thrombin proteolytic cleavage site is 

located between the Schistosoma japonicum glutathione S-transferase (GST) sequence 

and the tat insert. 

Cloning Site: BamHI–EcoRI. 

Cloning Vector: pGEX2T. 

Special 


Mutated HIV-2 Tat is expressed as a GST-Tat fusion protein, which can be easily 

expressed as described in the attached protocol. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Sandra Tong-Starksen. 

References: 


factor Sp1. Virology 238:221-230, 1997. 




REV: 11/08/2015 Page 1 of 4

NOTE: 




Limited to one aliquot per laboratory. 


Clone 

GST-Tat2 

(R81–84A) 

GST-Tat2 

(46–130) 

GST-Tat2 

(1–80) 

Cat. 

No. 

3642 

Description 

Two exons; substitution mutation of the basic residues 

(R81-84) to alanine in full-length tat2 cDNA. 

3643 Two exons; aa 1-45 deleted. 

3644 One exon; aa 81-130 deleted. 

Preparation and Purification of HIV-2 Tat from Bacterial Expression Systems 

Dr. Andrew P. Rice, Dr. Christine H. Herrmann, and Dr. Hyangshuk Rhim, Division of 

Molecular Virology, Baylor College of Medicine, One Baylor Plaza, Room 824D, Houston, 

TX 77030 

Reagents: 

LB-amp Medium 

Luria Broth containing 50 μg/ml ampicillin 

GST-Tat 

Catalog #3642–3644 

Expression Plasmids 

IPTG 

EBC Buffer 

EBC-DTT Buffer 

EBC-DTT-SDS 

Buffer 

Protease Inhibitors 

Lysozyme 

Glutathione 

Sepharose beads 

Thrombin Cleavage 

Buffer 

Human Thrombin 

3X Freeze Solution 

Other Reagents 

Isopropylthio-β-galactoside (BRL Catalog #5529). Make 100 mM 

stock solution using distilled water. 

50 mM Tris-HCl (pH 8.0), 120 mM NaCl, 0.5% NP-40 

EBC containing 5 mM DTT (make fresh) 

EBC-DTT buffer containing 0.075% SDS (make fresh) 

Prepare at the following final concentrations: Aprotinin (2 µg/ml), 

Leupeptin (1 βg/ml), PMSF (50 βg/ml). Stocks should be made 

fresh in 4 ml final volume (see step 6 below). 

16 mg/ml stock 

Pharmacia Catalog #17-0756-01 

50 mM Tris-HCl (pH 7.6), 20 mM KCl, 1 mM DTT 

Sigma catalog #T-0310 (supplied as a resuspension) 

60% glycerol containing 15 mM DTT (make fresh) 

SDS-PAGE Gel and buffers; Coomassie blue; protein standard 

markers 

Culture and Preparation of GST-Tat Extracts: 

1. Inoculate 50 ml of LB-amp medium with the glycerol stock of E. coli harboring 




REV: 11/08/2015 Page 2 of 4


the desired GST-Tat expression plasmid. 

2. Place the culture in a shaking 37°C incubator and leave overnight. 

3. The next day, dilute the culture 1:10 in LB-amp to give 500 ml final volume. 

Continue the incubation for 3 more hours. 

4. Add 0.5 ml of IPTG to give a 0.1 mM final concentration. Continue the incubation 

for 1.5 more hours. 

5. Transfer the culture to centrifuge tubes and centrifuge at 5000 rpm for 10 

minutes at 4°C. 

6. Pour the medium off and retain the bacterial pellet. Resuspend the pellet in 4 ml 

of EBC-DTT buffer containing protease inhibitors, and 2 mg/ml lysozyme, if 

desired. Note that lysozyme should not be used if the GST-Tat preparation is to 

be used in RNA binding experiments, as there is some concern that residual 

lysozyme may interfere with RNA binding. 

7. Place the tubes on ice for 15 minutes. 

8. Sonicate the tubes for 30 seconds, and repeat twice. Place the tubes on ice 

between the sonications. 

9. Transfer the solubilized bacteria to 1.5 ml microfuge tubes. Centrifuge at 12,000 

rpm for 15 minutes at 4°C using a microfuge. 

10. Transfer the supernatant to fresh tubes and make 1 ml aliquots. The 

supernatant can be stored at -70°C, and will remain stable for at least one year 

at this temperature. 

Estimation of Protein Concentration: 

1. Thaw the stock supernatant by placing the tube briefly at 37°C. 

2. Add 250 μl EBC-DTT Buffer to 50 μl of each E. coli extract. 

3. Add 25 μl of equilibrated glutathione-sepharose beads. [Equilibrate beads as 

follows: Pellet the beads (supplied by the manufacturer in 80% solution) by 

centrifuging at 12,000 rpm for 10 seconds in a microfuge. Pour off the 

supernatant. Add 500-1000 μl EBC-DTT, invert the tube several times (do not 

vortex). Pellet the beads as described above and repeat the washing. 

Resuspend the final bead pellet to give a 50% suspension in EBC-DTT.] 

4. Incubate the samples 15-30 minutes on a rocker placed in a cold room (4°C). 

5. Wash the beads by centrifuging for 10 seconds at 12,000 rpm in a microfuge, 

discard the supernatant, then add 500 μl of EBC-DTT-SDS buffer and invert the 

tube several times. Repeat the centrifugation and wash step one time. 

6. Resuspend the final pellet in volume of SDS-PAGE buffer. 

7. Load the samples onto a 15%DSD PAGE gel with 2.5 μg marker standards. If 

desired, load 1 μl of the unbound E. coli extracts. 

8. After completing the electrophoresis, stain the gel with Coomassie blue. 

9. Estimate the protein concentration by comparing the GST-Tat bands with 

marker standards and/or previous preparations. 

Thrombin Digestion: 

1. Transfer the beads into a 15 ml conical tube and add 50-fold volume of thrombin 




REV: 11/08/2015 Page 3 of 4


cleavage buffer. Centrifuge the sample at 3000 rpm for 3 minutes at room 

temperature. 

2. Discard the supernatant and resuspend the pellet in 100 μl of thrombin cleavage 

buffer per ml of E. coli lysate + 10 μl (equal to 5 units or 1.5 μg) of the human 

thrombin. Incubate at room temperature for 1 hour with gentle mixing. 

3. To elute the cleaved Tat protein from the GST-bound sepharose, incubate the 

beads at 37°C for 3 minutes then briefly centrifuge the sample at 12,000 rpm 

for 30 seconds at room temperature using a microfuge. 

4. Collect the eluate and repeat step 3 two times. 

5. Pool the protein eluates. Adjust their concentrations in 1X freeze solution to give 

final concentrations of 20% glycerol and 7 mM DTT. Store the preparations at 

-70°C. 




REV: 11/08/2015 Page 4 of 4

DATA SHEET 

Reagent: GST-Tat2 (46-130) 


Lot Number: 1 

Release 

Category: 

A 


Description: 






Special 




Recommended 

Storage: 

-70°C. 


References: 






REV: 11/08/2015 Page 1 of 4

NOTE: 






Clone 

GST-Tat2 

(R81–84A) 

GST-Tat2 

(46–130) 

GST-Tat2 

(1–80) 

Cat. 

No. 

3642 

Description 








TX 77030 

Reagents: 

LB-amp Medium 


GST-Tat 

Catalog #3642–3644 


IPTG 

EBC Buffer 


EBC-DTT-SDS 

Buffer 


Lysozyme 

Glutathione 



Buffer 


















markers 






REV: 11/08/2015 Page 2 of 4















































REV: 11/08/2015 Page 3 of 4



temperature. 










-70°C. 




REV: 11/08/2015 Page 4 of 4

DATA SHEET 

Reagent: 



Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 2 84D 


Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: GST-Tat 2 99R D8/47 


Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: GST-Tat 2 99R D8/33 


Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 2 99R 


Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

GST-Tat 2 99R 8182A 


Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 



Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 



Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: GST-Tat2 (1-80) 


Lot Number: 1 

Release 

Category: 

A 


Description: 






Special 




Recommended 

Storage: 

-70°C. 


References: 






REV: 11/08/2015 Page 1 of 4

NOTE: 






Clone 

GST-Tat2 

(R81–84A) 

GST-Tat2 

(46–130) 

GST-Tat2 

(1–80) 

Cat. 

No. 

3642 

Description 








TX 77030 

Reagents: 

LB-amp Medium 


GST-Tat 

Catalog #3642–3644 


IPTG 

EBC Buffer 


EBC-DTT-SDS 

Buffer 


Lysozyme 

Glutathione 



Buffer 


















markers 






REV: 11/08/2015 Page 2 of 4















































REV: 11/08/2015 Page 3 of 4



temperature. 










-70°C. 




REV: 11/08/2015 Page 4 of 4

DATA SHEET 

Reagent: 

GST-Tat 2 99R TK 


Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 





REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 



Lot Number: 2 

Release 

Category: 

B 


Description: 










Special 









Recommended 

Storage: 

-90°C. 

Contributor: Dr. Andrew Rice 




REV: 11/08/2015 Page 1 of 2

References: 







NOTE: 







REV: 11/08/2015 Page 2 of 2

IL-2 >> All

DATA SHEET 

Reagent: 

pBC12 (CMV) IL-2 




Provided: 5 μg purified plasmid DNA (1 μg/μL). 

Description: Annonations for pBC12 (CMV) IL-2 

Cloning Site: 

The insert is cloned into HIND III/BAMH I sites of pBC12/CMV. Insert size is 683bp. The 

size of the cloning vector plus insert is 4732bp. 

Cloning Vector: pBC12/CMV (ampicillin resistant). 

Special 


Expresses secreted human IL-2. 

Recommended 

Storage: 

-20°C 


References: 

Cullen BR. Trans-activation of human immunodeficiency virus occurs via a bimodal 

mechanism. Cell 1986 Sep 26; 46 (7): 973-82. [Abstract] 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pBC12 (CMV) IL-2 

(Cat#11416) from Dr. Bryan R. Cullen." Also include the reference cited above in any 

publications. 






REV: 11/08/2015 Page 1 of 2









REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pLiv7 




Provided: 100ng/µl plasmid DNA in TE buffer, 10µl/vial 

Description: 

Expression vector containing the hepatic control region of the human apolipoprotein E3 

gene (originally described and constructed by John M. Taylor, see attached map). 

map 


pBS-SK. The size of the vector including the insert is approximately 8.5Kb. The size of 

the insert is approximately 5.9Kb. Cloning sites are shown in the map below. 

Special 


Drives liver-specific gene expression. 

Recommended 

Storage: 

-20°C 

Contributor: Dr. Simon Hui 

References: 

Miyake J. H., Doung X. T., Strauss W., Moore G. L., Castellani L. W., Curtiss L. K., 

Taylor J. M. and Davis R. A. Increased Production of Apolipoprotein B-containing 

Lipoproteins in the Absence of Hyperlipidemia in Transgenic Mice Expressing 

Cholesterol 7 - Hydroxylase. J. Biol. Chem. 276: 23304-23311, 2001. 

Simonet W. S., Bucay N., Lauer S. J., and Taylor J. M. A far-downstream 

hepatocyte-specific control region directs expression of the linked human 

apolipoprotein E and C-I genes in transgenic mice. J. Biol. Chem. 268: 8221-8229, 

1993. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH from Dr. Simon 

Hui: pLiv." Also include the references cited above in any publications. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pFT-1-LEDGF 


Lot Number: 1 


Provided: 10ul, 1ug of plasmid DNA in TE buffer, propagate in XL1-Blue or DH5-α 

Description: 

Codes for Human LEDGF/p75; expression in BL21(DE3) is induced by addition of IPTG. 

LEDGF/p75 is a host factor that aids HIV proviral integration. 

Sequence 

Plasmid Map 

Special 


Vector for expressing His-tagged human LEDGF/p75 in bacteria under control of the T7 

promoter. The His-tag can be removed using PreScission Protease. 

Contributor: Dr. Alan Engelman 

References: Vandegraaff N, et. al. Virology. 2006 Mar 15;346(2):415. 

Click here for abstract. 

NOTE: 

Click here for sequence document. 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pFT-1-LEDGF 

(Cat#11396) from Dr. Alan Engelman." Also include the reference cited above in any 

publications. 

Requests from commercial organizations should be directed to Nancy Grodin 

at Nancy_Grodin@dfci.harvard.edu 




REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

CMVR VRC03 H 




Provided: 1.4 μg/μl plasmid DNA in TE, pH 8.0. 

Description: 

The leader, variable and constant regions of VRC03 heavy chain are expressed under 

control of the CMV promoter. 

Cloning Vector: CMV/R 

Special 


The plasmid expresses the IgG heavy chain of the HIV-1 gp120 mAb, VRC03. With 

catalog #12038, CMVR VRC03 L, the VRC03 mAb (catalog #12032) can be expressed in 

293F cells and purified using a protein-A column (see protocol and references). VRC03 

is a broadly neutralizing HIV-1 antibody; active against all major subtypes. 

Sequence File 

Protocol 

Recommended 

Storage: 

−20°C 

Contributor: Xueling Wu, Zhi-Yong Yang, Yuxing Li, Gary Nabel, and John Mascola. 

References: 

Wu X, Yang ZY, Li Y, Hogerkorp CM, Schief WR, Seaman MS, Zhou T, Schmidt SD, Wu 

L, Xu L, Longo NS, McKee K, O'Dell S, Louder MK, Wycuff DL, Feng Y, Nason M, 

Doria-Rose N,Connors M, Kwong PD, Roederer M, Wyatt RT, Nabel GJ, Mascola JR. 

Rational design of envelope identifies broadly neutralizing human monoclonal antibodies 

to HIV-1. Science 329(5993):856-61, 2010. 

Reference for CMV/R vector: 

Barouch DH, Yang ZY, Kong W-P, Korioth-Schmitz B, Sumida SM, Truitt DM, Kishko MG, 




REV: 11/08/2015 Page 1 of 2


Arthur JC, Miura A, Mascola JR, Letvin NL, Nabel GJ. A human T-cell leukemia virus type 

1 regulatory element enhances the immunogenicity of human immunodeficiency virus 

type 1 DNA vaccines in mice and nonhuman primates. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: CMVR VRC03 H, 

from Dr. John Mascola." Also include the reference cited above in any publications. 


reagent must contact: The Office of Technology Development, NIAID, 6610 

Rockledge Drive, Suite 2800, MSC 6606, Bethesda, MD, 20892-6606, Tel: 

301-496-2644, Fax: 301-402-7123, before the reagent can be released. 

Patent Pending. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

CMVR VRC03 L 




Provided: 1.2 μg/μl plasmid DNA in TE, pH 8.0. 

Description: 

The leader, variable and constant regions of VRC03 light chain are expressed under 


Cloning Site: See sequence. 


Special 






Sequence File 

Protocol 

Recommended 

Storage: 

−20°C 





REV: 11/08/2015 Page 1 of 2

References: 





to HIV-1. Science 329(5993):856-61, 2010. 






NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: CMVR VRC03 L, 










REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

10E8 mAb Heavy chain expression vector (CMVR) 



Release 

Category: 

C 

Provided: 5 μg of plasmid DNA. 

Description: 

The heavy chain expression vector for the broadly neutralizing antibody, 10E8. The 

expression vector codes for a signal peptide sequence, variable and constant regions of 

IgG1 heavy chain expressed under control of the HCMV (human cytomegalovirus 

immediate-early) promoter. 


Size of insert is about 500 bp. The vector is the same as VRC01 (Cat# 12035). The 

sequence and map are provided in the link below. The vector's selection drug is 

Kanamycin at 50μg/ml. 

Special 


10E8 is a broadly neutralizing HIV-1 antibody against all major subtypes. This plasmid 

expresses the IgG1 heavy chain of the HIV-1 gp41 mAb 10E8. With 10E8 light chain 

plasmid (Cat# 12291), or the 7H6 light chain plasmid (Cat# 12293), the 10E8 mAb 

(Cat# 12294) or the 7H6 mAb (Cat# 12295) can be expressed in 293T cells and purified 

using a protein-A column (please see references). 10E8 tends to precipitate at 4°C or 

after -20°C storage, please warm it up at 37°C for 1 hour before use. 

GenBank #JX645769. 

Sequence and vector map 

Recommended 

Storage: 

-20°C 

Contributor: Jinghe Huang, Leo Laub, and Mark Connors 




REV: 11/08/2015 Page 1 of 2

References: Jinghe Huang, Gilad Ofek, Leo Laub, Mark K. Louder, Nicole A. Doria-Rose, Nancy S. 

Longo, Hiromi Imamichi, Robert T. Bailer, Bimal Chakrabarti, Shailendra K. Sharma S. 

Munir Alam, Tao Wang, Yongping Yang, Baoshan Zhang, Stephen A. Migueles, Richard 

Wyatt, Barton F. Haynes, Peter D. Kwong, John R. Mascola and Mark Connors. Broad and 

potent neutralization of HIV-1 by a gp41-specific human antibody. Nature (2012) 

doi:10.1038/ nature11544 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: mAb 10E8 heavy 

chain, from Dr. Mark Connors." Also include the reference cited above in any 

publications. Scientists at for-profit institutions or who intend commercial use of this 

reagent must contact: The Office of Technology Development, NIAID, 6610 Rockledge 

Drive, Suite 2800, MSC 6606, Bethesda, MD, 20892-6606, Tel:301-496-2644, Fax: 

301-402-7123, before the reagent can be released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

10E8 mAb Light chain expression vector (CMVR) 



Release 

Category: 

C 

Provided: 5.0 μg of plasmid DNA. 

Description: 

The light chain expression vector for the broadly neutralizing antibody, 10E8. The 


IgG light (lambda) chain expressed under control of the HCMV (human cytomegalovirus 



The size of the insert is about 400 bp. The vector is the same as VRC01 light chain (Cat# 

12036). The sequence and map are provided in the link below. The vector's selection 

drug is Kanamycin at 50 µg/ml. 

Special 


10E8 is a broadly neutralizing HIV-1 antibody against all major subtypes. This plasmid 

expresses the IgG1 heavy chain of the HIV-1 gp41 mAb 10E8. With 10E8 heavy chain 

plasmid (Cat# 12290), or the 7H6 light chain plasmid (Cat# 12293), the 10E8 mAb 

(Cat# 12294) or the 7H6 mAb (Cat# 12295) can be expressed in 293T cells and purified 

using a protein-A column (please see references). 10E8 tends to precipitate at 4°C or 

after -20°C storage, please warm it up at 37°C for 1 hour before use. 



Recommended 

Storage: 

-20°C 





REV: 11/08/2015 Page 1 of 2






doi:10.1038/ nature11544 

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

7H6 mAb Light chain expression vector (CMVR) 



Release 

Category: 

C 

Provided: 5 μg of plasmid DNA. 

Description: 

The light chain expression vector for the broadly neutralizing antibody, 7H6. The 


IgG light (lambda) chain expressed under control of the HCMV (human cytomegalovirus 



The size of the insert is about 400 bp. The vector is the same as VRC01 light chain (Cat# 

12036). The sequence and map are provided in the link below. The vector's selection 

drug is Kanamycin at 50 µg/ml. 

Special 


7H6 is a broadly neutralizing HIV-1 antibody against all major subtypes. This plasmid 

expresses the IgG (lambda) light chain of the HIV-1 gp41 mAb 7H6. The 7H6 antibody 

has the same heavy chain as 10E8 (Cat #12290). The 7H6 mAb (Cat# 12294) can be 

expressed in 293T cells and purified using a protein-A column (please see references). 

7H6 tends to precipitate at 4°C or after -20°C storage, please warm it up at 37°C for 1 

hour before use. 



Recommended 

Storage: 

-20°C 





REV: 11/08/2015 Page 1 of 2






doi:10.1038/ nature11544 

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

35O22 mAb Heavy chain expression vector (CMVR) 



Release 

Category: 

C 

Provided: 5 µg of purified DNA at 1 mg/mL. 

Description: 

The heavy chain expression vector for the broadly neutralizing antibody, 35O22. The 


IgG1 heavy chain expressed under control of the HCMV (human cytomegalovirus 



The vector is the same as VRC01 mAb Heavy chain expression vector (CMVR) (Cat# 

12035). The plasmid sequence is provided in the link below. The vector's selection drug is 

Kanamycin at 50 µg/ml. 

Sequence File 

Special 


35O22 is a broadly neutralizing HIV-1 antibody against all major subtypes that binds to 

gp120 and gp41. This plasmid expresses the IgG1 heavy chain of mAb 35O22 (Cat# 

12586). With the 35O22 mAb Light chain expression vector (Cat# 12585), 35O22 can be 

expressed in 293T cells and purified using a protein-A column (please see references). 

35O22 tends to precipitate at 4°C or after -20°C storage, please warm it up at 37°C for 1 

hour before use. 

Recommended 

Storage: 

-20°C 

Contributor: Jinghe Huang and Mark Connors 




REV: 11/08/2015 Page 1 of 2

References: 

Jinghe Huang, Byong H. Kang, Marie Pancera, Jeong Hyun Lee, Tommy Tong, Yu Feng, 

Hiromi Imamichi, Ivelin S. Georgiev, Gwo-Yu Chuang, Aliaksandr Druz, Nicole A. 

Doria-Rose, Leo Laub, Kwinten Sliepen, Marit J. van Gils, Alba Torrents de la Peña, Ronald 

Derking, Per-Johan Klasse, Stephen A. Migueles, Robert T. Bailer, Munir Alam, Pavel 

Pugach, Barton F. Haynes, Richard T. Wyatt, Rogier W. Sanders, James M. Binley, Andrew 

B. Ward, John R. Mascola, Peter D. Kwong & Mark Connors. Broad and potent HIV-1 

neutralization by a human antibody that binds the gp41–gp120 interface. Nature (2014) 

doi: 10.1038/nature13601 

For antibody purification, follow the protocol for 10E8. 

Jinghe Huang, Gilad Ofek, Leo Laub, Mark K. Louder, Nicole A. Doria-Rose, Nancy S. 





doi:10.1038/ nature11544 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: Cat# 12584 mAb 

35O22 heavy chain expression vector (CMVR), from Drs. Jinghe Huang and Mark 

Connors." Also include the reference cited above in any publications. Scientists at 

for-profit institutions or who intend commercial use of this reagent must contact: The 

Office of Technology Development, NIAID, 6610 Rockledge Drive, Suite 2800, MSC 6606, 

Bethesda, MD, 20892-6606, Tel:301-496-2644, Fax: 301-402-7123, before the reagent 

can be released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

35O22 mAb Light chain expression vector (CMVR) 



Release 

Category: 

C 

Provided: 5 µg of purified DNA at 1 mg/mL. 

Description: 

The light chain expression vector for the broadly neutralizing antibody, 35O22. The 


IgG1 light chain expressed under control of the HCMV (human cytomegalovirus 



The vector is the same as VRC01 mAb Heavy chain expression vector (CMVR) (Cat# 

12035). The plasmid sequence is provided in the link below. The vector's selection drug is 

Kanamycin at 50 µg/ml. 

Sequence File 

Special 


35O22 is a broadly neutralizing HIV-1 antibody against all major subtypes that binds to 

gp120 and gp41. This plasmid expresses the IgG1 light chain of mAb 35O22 (Cat# 

12586). With 35O22 heavy chain plasmid (Cat# 12584) 35O22 can be expressed in 293T 

cells and purified using a protein-A column (please see references). 35O22 tends to 

precipitate at 4°C or after -20°C storage, please warm it up at 37°C for 1 hour before use. 

Recommended 

Storage: 

-20°C 

Contributor: Jinghe Huang and Mark Connors 




REV: 11/08/2015 Page 1 of 2

References: 

Jinghe Huang, Byong H. Kang, Marie Pancera, Jeong Hyun Lee, Tommy Tong, Yu Feng, 

Hiromi Imamichi, Ivelin S. Georgiev, Gwo-Yu Chuang, Aliaksandr Druz, Nicole A. 

Doria-Rose, Leo Laub, Kwinten Sliepen, Marit J. van Gils, Alba Torrents de la Peña, Ronald 

Derking, Per-Johan Klasse, Stephen A. Migueles, Robert T. Bailer, Munir Alam, Pavel 

Pugach, Barton F. Haynes, Richard T. Wyatt, Rogier W. Sanders, James M. Binley, Andrew 

B. Ward, John R. Mascola, Peter D. Kwong & Mark Connors. Broad and potent HIV-1 

neutralization by a human antibody that binds the gp41–gp120 interface. Nature (2014) 

doi: 10.1038/nature13601 

For antibody purification, follow the protocol for 10E8. 

Jinghe Huang, Gilad Ofek, Leo Laub, Mark K. Louder, Nicole A. Doria-Rose, Nancy S. 





doi:10.1038/ nature11544 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: Cat# 12585 mAb 

35O22 light chain, from Drs. Jinghe Huang and Mark Connors." Also include the reference 

cited above in any publications. Scientists at for-profit institutions or who intend 

commercial use of this reagent must contact: The Office of Technology Development, 

NIAID, 6610 Rockledge Drive, Suite 2800, MSC 6606, Bethesda, MD, 20892-6606, 

Tel:301-496-2644, Fax: 301-402-7123, before the reagent can be released. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

CMVR VRC01 H 




Provided: 5μg at 1.0μg/μl purified DNA 

Description: 



Cloning Site: See plasmid map and sequence. 


Special 


The plasmid expresses the IgG heavy chain of the HIV-1 gp120 MAb, VRC01. With 

catalog #12036, CMVR VRC01 L, the VRC01 mAb (Catalog# 12033)can be expressed in 

293F cells and purified using a protein-A column (see protocol and references).VRC01 is 

a broadly neutralizing HIV-1 antibody; active against all major subtypes. 

Sequence File 

Protocol 

Recommended 

Storage: 

−20°C 

Contributor: Xueling Wu, Zhi-Yong Yang, Yuxing Li, Gary Nabel, and John Mascola 




REV: 11/08/2015 Page 1 of 2

References: 





to HIV-1. Science 329(5993):856-61, 2010. 






NOTE: 












REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

CMVR VRC01 L 




Provided: 5μg at 1.0 μg/μl purified DNA 

Description: 




Special 


The plasmid expresses the IgG light chain of the HIV-1 gp120 MAb, VRC01. With 

catalog #12035, CMVR VRC01 H, the VRC01 mAb (catalog# 12033) can be expressed 

in 293F cells and purified using a protein-A column (see protocol and references). 

VRC01 is a broadly neutralizing HIV-1 antibody; active against all major subtypes. 

Sequence File 

Protocol 

Recommended 

Storage: 

−20°C 

Contributor: Xueling Wu, Zhi-Yong Yang, Yuxing Li, Gary Nabel, and John Mascola 

References: 





to HIV-1. Science 329(5993):856-61, 2010. 






REV: 11/08/2015 Page 1 of 2





NOTE: 












REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

CMVR VRC03 H 





Description: 




Special 








Protocol 

Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 





to HIV-1. Science 329(5993):856-61, 2010. 






NOTE: 












REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

CMVR VRC03 L 





Description: 




Special 








Protocol 

Recommended 

Storage: 






REV: 11/08/2015 Page 1 of 2

References: 





to HIV-1. Science 329(5993):856-61, 2010. 






NOTE: 












REV: 11/08/2015 Page 2 of 2

SIV >> Tat

DATA SHEET 

Reagent: 

pCEP4SIVtat 



Release 

Category: 

C 

Provided: 1 vial containing 5μg of purified DNA in TE buffer 

Description: 

SIVmne Tat expression driven by CMV IE promoter in vector pCEP4. SIVmne mRNA 

coding for Tat was amplified from M. fascicularis PBMC using RT-PCR with specific 

primers beginning at the AUG initiation codon and ending at the UAG stop codon. The 

SIV tat gene is 396 bases in length, comprised of exon 1 (1-296) and exon 2 (297-396). 

The complete SIV Tat gene PCR product was cloned into pCRII (Invitrogen) and then 

subcloned into pCEP4 (Invitrogen) between the XhoI and BamHI sites. 

pCEP4SIVTat prokaryotic/eukaryotic vector description 

Special 


Expresses SIV Tat in eukaryotic cells after transfection. Can be made semi-permanent 

through maintenance of episomal plasmid by selection in hygromycin B. Expression of Tat 

can be indirectly measured by co-transfection with SIV or HIV LTR-ß-gal plasmid and 

measurement of ß-gal levels. Expression of Tat can be enhanced by cotransfection with 

plasmid expressing CMV IE protein (Peter Barry, UC, Davis). Reagent 293T-CEP4SIVtat 

(Cat# 8123) is a semi-permanent 293T cell line that constitutively expressed SIV Tat 

from this plasmid. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Richard Grant. 

References: 

Grant RF, Stevens Y, Wright N, Agy M, Thouless M, Morton W. Stable SIV Tat Cell Lines 

Increase Expression Of Transfected Genes. J Med Primatol 28:(abstract), 1999. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pCEP4SIVtat from 

Dr. Richard Grant." Also include the reference cited above in any publications. 

Requests from commercial organizations should be directed to Ariadna A. 

Santander, Program Operations Coordinator, Office of Technology Licensing, 

University of Washington, 1107 N.E 45th Street, Suite 200, Seattle, WA 98195. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pCEP4SIVtat 


Lot Number: 20 Aug 2002 

Release 

Category: 

C 

Provided: 1 vial of transformed E. coli JM109 in LB with 15% glycerol. 

Description: 

SIVmne Tat expression driven by CMV IE promoter in vector pCEP4. SIVmne mRNA 

coding for Tat was amplified from M. fascicularis PBMC using RT-PCR with specific 

primers beginning at the AUG initiation codon and ending at the UAG stop codon. The 

SIV tat gene is 396 bases in length, comprised of exon 1 (1-296) and exon 2 (297-396). 

The complete SIV Tat gene PCR product was cloned into pCRII (Invitrogen) and then 

subcloned into pCEP4 (Invitrogen) between the XhoI and BamHI sites. 

pCEP4SIVTat prokaryotic/eukaryotic vector description 

Special 


Expresses SIV Tat in eukaryotic cells after transfection. Can be made semi-permanent 

through maintenance of episomal plasmid by selection in hygromycin B. Expression of Tat 

can be indirectly measured by co-transfection with SIV or HIV LTR-ß-gal plasmid and 

measurement of ß-gal levels. Expression of Tat can be enhanced by cotransfection with 

plasmid expressing CMV IE protein (Peter Barry, UC, Davis). Reagent 293T-CEP4SIVtat 

(Cat# 8123) is a semi-permanent 293T cell line that constitutively expressed SIV Tat 

from this plasmid. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Richard Grant. 

References: 

Grant RF, Stevens Y, Wright N, Agy M, Thouless M, Morton W. Stable SIV Tat Cell Lines 

Increase Expression Of Transfected Genes. J Med Primatol 28:(abstract), 1999. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pCEP4SIVtat from 

Dr. Richard Grant." Also include the reference cited above in any publications. 

Requests from commercial organizations should be directed to Ariadna A. 

Santander, Program Operations Coordinator, Office of Technology Licensing, 

University of Washington, 1107 N.E 45th Street, Suite 200, Seattle, WA 98195. 




REV: 11/08/2015 Page 2 of 2

SIV >> Vpr

DATA SHEET 

Reagent: 

SIV pGEX-KG vpr 




Provided: 1 vial ampicillin-resistant DH5 bacteria. 

Description: 

Contains a PCR-amplified EcoRI-HindIII fragment encoding the entire SIVsmPBj1.9 vpr 

open reading frame cloned into pGEX-KG, a prokaryotic expression vector. 

Cloning Vector: pGEX-KG, a prokaryotic expression vector. 

Special 


Contains a 300 bp vpr insert. Expresses high levels of Vpr as a glutathione S-transferase 

fusion product after IPTG induction. The vpr open reading frame and vector junctions 

have been sequenced to insure fidelity and maintenance of the proper frame. The protein 

is easily purified on GST columns, and efficiently isolated by thrombin cleavage. 

Recommended 

Storage: 

-70°C. 

Contributor: Dr. Margaret Newman and Dr. Beatrice Hahn. 

References: 

Newman MA, McPherson SA, Hahn BH. Polyclonal rabbit antisera that detect the vpr 

protein of SIVmac and SIVsm on immunoblots of purified proteins. AIDS Res Hum 

Retroviruses 11:405-408, 1995. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pGEX-KG vpr 

from Dr. Margaret Newman and Dr. Beatrice Hahn." Also include the reference cited 


Scientist at for-profit institutions or who intend commercial use of Release 

Category C reagent (SIV pGEX-KG vpr, Cat# 2749), must contact Dr. Leona C. 




REV: 11/08/2015 Page 1 of 2

Category C reagent (SIV pGEX-KG vpr, Cat# 2749), must contact Dr. Leona C. 

Fitzmaurice, UAB Research Foundation, Office of Intellectual Property 

Management, AB770, 1530 3rd Ave S., Birmingham, AL, 35294-0107, Tel: 

205-934-7868, Fax: 205-934-5427, Email:fitzmaur@uab.edu, before the 





REV: 11/08/2015 Page 2 of 2

TRIM5α >> All

DATA SHEET 

Reagent: 

pLPCX-TRIM5α hu -HA (human) 

Catalog 

Number: 

10074 


Release 

Category: 

C 

Provided: 

5.0 µg of plasmid DNA in 5.0 µl of 10 mM Tris-HCl, pH 7.5 buffer. DH5α competent cells 

(Invitrogen) should be used for propagation. 

Description: 

The human TRIM5α open reading frame was amplified from a kidney cDNA library 

(Clontech). The predicted sequence of the TRIM5α differs in three amino acid residues from 

NCBI RefSeq NM-033034 (this sequence contains an isoleucine at position 76, a leucine at 

position 130, and a glutamine at position 136). 

The primers used for cloning HA-tagged Human TRIM5α into pLPCX are: 

Forward: 

gcggaattcgccaccATGTACCCATACGA 

CGTCCCAGACTACGCTGGCGGCgcttctggaatcctggttaatgtaaag 

Reverse: 

ccatcgatggcTCAAGCGTAGTCTGGGAC GTCGTATGGGTAGCCGCCagagcttggtgagcacagagtcatg 

Cloning Site: 

The insert is cloned between the EcoRI and CIaI sites. The EcoRI site lies at the 5’-end of 

the insert and the CIaI site at the 3’-end. The size of the insert is approximately 1.5 kb. 


The cloning vector is pLPCX (available from Clontech). pLPCX is an MLV retroviral vector. It 

contains a puromycin resistance gene and expression of TRIM5α is driven by the CMV 

promoter. The size of the vector containing the insert is approximately 6.8 kb. pLPCX is 

ampicillin resistant. 

Recommended 

Storage: 

-20°C 




REV: 11/08/2015 Page 1 of 2

Contributor: Drs. Joseph Sodroski and Matt Stremlau 

References: 

Stremlau, M et al. The cytoplasmic body component TRIM5α restricts HIV-1 infection in Old 

World monkeys. Nature 427: 848-853, 2004 

NOTE: 


the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pLPCX-TRIM5α hu -HA 

(human) from Drs. Joseph Sodroski and Matt Stremlau." Also include the reference cited 

above in any publications. Scientists at for-profit institutions or who intend 

commercial use of this reagent must contact Nancy Grodin at email address 

nancy_grodin@dfci.harvard.edu and specify in the email the name of the reagent 

and a description of the intended use of the reagent. Patent Pending. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pLPCX-TRIM5α rh (rhesus monkey) 




Provided: 

1.0 µg of plasmid DNA in 10 µl of 10 mM Tris-HCl, pH 7.5 buffer. DH5α competent cells 


Description: 

The Rhesus TRIM5α was cloned from cDNA prepared from primary Rhesus lung 

fibroblast cells. PCR was used to clone the cDNA into the vector. 

pLPCX is an MLV retroviral vector. It contains a puromycin resistance gene and 

expression of TRIM5α is driven by the CMV promoter. 

Protein and DNA sequence information available. Click here to see the sequence file 

Cloning Site: 

The insert is cloned between the EcoRI and CIaI sites. The EcoRI site lies at the 5’-end 

of the insert and the CIaI site at the 3’-end. The size of the insert is approximately 1.5 

kb. 


The cloning vector is pLPCX (available from Clontech). The size of the vector containing 

the insert is approximately 6.8 kb. pLPCX is ampicillin resistant. 

Special 


Cloning Strategy: The cloning vector is pLPCX (available from Clontech). 

Recommended 

Storage: 

-20°C. 





REV: 11/08/2015 Page 1 of 2

References: 

Stremlau, M et al. The cytoplasmic body component TRIM5a restricts HIV-1 infection in 

Old World monkeys. Nature 427: 848-853, 2004, 

NOTE: 



pLPCX-TRIM5a rh (rhesus monkey) from Drs. Joseph Sodroski and Matt Stremlau." Also 



reagent must contact Nancy Grodin at email address 

nancy_grodin@dfci.harvard.edu and specify in the email the name of the 

reagent and a description of the intended use of the reagent. Patent Pending. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pLPCX-TRIM5α hu (human) 



Release 

Category: 

C 

Provided: 

1.0 µg plasmid DNA in 10 µl of 10 mM Tris-HCl, pH 7.5 buffer. DH5α competent cells 


Description: 

The human TRIM5α open reading frame was amplified from a kidney cDNA library 

(Clontech). The predicted sequence of the TRIM5α differs in three amino acid residues 

from NCBI RefSeq NM-033034 (this sequence contains an isoleucine at position 76, a 

leucine at position 130, and a glutamine at position 136). 

pLPCX is an MLV retroviral vector. It contains a puromycin resistance gene and expression 

of TRIM5α is driven by the CMV promoter. 

Protein and DNA sequence information available. 

Click here to see the sequence file 

Cloning Site: 

The insert is cloned between the EcoRI and CIaI sites. The EcoRI site lies at the 5’-end of 

the insert and the CIaI site at the 3’-end. The size of the insert is approximately 1.5 kb. 

Cloning Vector: The cloning vector is pLPCX (available from Clontech). 

Special 


Cloning Strategy: The cloning vector is pLPCX (available from Clontech). The size of the 

vector containing the insert is approximately 6.8 kb. pLPCX is ampicillin resistant. 

Recommended 

Storage: 

-20°C. 





REV: 11/08/2015 Page 1 of 2

References: 


Old World monkeys. Nature 427: 848-853, 2004 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pLPCX-TRIM5a hu 

(human) from Drs. Joseph Sodroski and Matt Stremlau." Also include the reference cited 

above in any publications. Scientists at for-profit institutions or who intend 

commercial use of this reagent must contact Nancy Grodin at email address 

nancy_grodin@dfci.harvard.edu and specify in the email the name of the reagent 

and a description of the intended use of the reagent. Patent Pending. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pLPCX-TRIM5α rh -HA (rhesus monkey) 



Release 

Category: 

C 

Provided: 

5.0 μg of plasmid DNA in 5.0 μl of 10 mM Tris-HCl, pH 7.5 buffer. DH5α competent cells 


Description: 

The Rhesus TRIM5α was cloned from cDNA prepared from primary Rhesus lung fibroblast 

cells. PCR was used to clone the cDNA into the vector. The primers used for cloning 

HA-tagged Rhesus TRIM5a into pLPCX are: 

Forward: 

gcggaattcgccaccATGTACCCATACGACGTCCCAGACTACGCT 

GGCGGC gcttctggaatcctggttaatgtaaag 

Reverse: 

ccatcgatggcTCAAGCGTAGTCTGGGACGTCGTATGGGTAGCCGCC 

agagcttggtgagcacagagtcatg 

pLPCX is an MLV retroviral vector. It contains a puromycin resistance gene and expression 

of TRIM5a is driven by the CMV promoter. Protein and DNA sequence information 

available. 

Click here to see the sequence file. 

Cloning Site: 

The insert is cloned between the EcoRI and CIaI sites. The EcoRI site lies at the 5′-end of 

the insert and the CIaI site at the 3′-end. The size of the insert is approximately 1.5 kb. 

Special 


Cloning Strategy: The cloning vector is pLPCX (available from Clontech). The size of the 

vector containing the insert is approximately 6.8 kb. pLPCX is ampicillin resistant. 




REV: 11/08/2015 Page 1 of 2

Recommended 

Storage: 

-20°C 

Contributor: Drs. Joseph Sodroski and Matt Stremlau. 

References: 


Old World monkeys. Nature 427: 848-853, 2004 

NOTE: 



pLPCX-TRIM5arh-HA (rhesus monkey) from Drs. Joseph Sodroski and Matt Stremlau." 

Also include the reference cited above in any publications. 


must contact Nancy Grodin at email address nancy_grodin@dfci.harvard.edu and 

specify in the email the name of the reagent and a description of the intended 

use of the reagent. Patent Pending. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pMIG-TRIMCyp 




Provided: 

The reagent is plasmid DNA diluted in TE buffer at a concentration of 1.0 μg/μl. The 

sample size is 4.0 μg (4 μl). The plasmid was amplified in DH5α. 

Description: 

Owl monkey TRIMCyp cDNA in pMIG, an MSCV-based retroviral expression vector. 

Expression is driven by the MSCV LTR and the cDNA is expressed as a fusion to an 

IRES-GFP cassette, allowing identification of transduced cells by their GFP expression. 

Cells expressing TRIMCyp are resistant to HIV-1 infection. 

Cloning Site: 

The approximately 1.5 kb insert was cloned between the Xho1 to EcoR1 restriction 

sites. 

Cloning Vector: The cloning vector is pMIG. The size of the cloning vector including the insert is ~8 kb. 

Recommended 

Storage: 

-20°C 

Contributor: Dr. David Sayah and Dr. Jeremy Luban. 

References: 

David M. Sayah, Elena Sokolskaja, Lionel Berthoux and Jeremy Luban. Cyclophilin A 

retrotransposition into TRIM5 explains owl monkey resistance to HIV-1. Nature 

430:569-573, 2004. 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pMIG-TRIMCyp 

from Dr. David Sayah and Dr. Jeremy Luban." Also include the reference cited above 

in any publications. 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pcGNM2/TSG-F (TSG101) 




Provided: 5 μg purified plasmid DNA at 0.8 μg/μL in TE. 

Description: 

Full-length TSG101 expression vector. Constructed by transferring the TSG101 coding 

region from plasmid pGST2/TSG5 (provided by Z. Sun). For more details, see 

Goila-Gaur et.al. (2003). 

Cloning Site: XbaI and BamHI (Insert size, 1196bp). 

Cloning Vector: pcGNM2 (The size of the cloning vector, including insert, is 6230bp). 

Special 


Over expression of pcGNM2/TSG-F inhibits HIV-1 budding. 

Sequence 

Plasmid Map 

Recommended 

Storage: 

4°C or lower. 

Contributor: Dr. Eric Freed 

References: 

Goila-Gaur R, Demirov DG, Orenstein JM, Ono A, Freed EO. Defects in human 

immunodeficiency virus budding and endosomal sorting induced by TSG101 over 

expression. J Virol. 2003, 77 (11); 6507-19. 

Demirov DG, Ono A, Orenstein JM, Freed EO. Over expression of the N-terminal 

domain of TSG101 inhibits HIV-1 budding by blocking late domain function. Proc. 

Natl. Acad. Sci. USA, 2002, 99 (2); 955-60. 




REV: 11/08/2015 Page 1 of 2

Natl. Acad. Sci. USA, 2002, 99 (2); 955-60. 

NOTE: 



pcGNM2/TSG-F (Cat#11483) from Dr. Eric Freed.” Also include the references cited 






the reagent. 




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pHEF-VSVG 




Provided: 5 μg plasmid DNA, 0.8 μg/μL, in TE. Propagate in DH5α, Top-10, HB101, or STBL2. 

Description: 

VSV-G envelope eukaryotic expression vector. PCR-amplified VSV-G sequence with a 

preferred eukaryotic translational initiation codon (-CCACC-ATG) driven by a strong 

human elongation factor alpha promoter. 

DNA Transfection for Production of Recombinant Lentiviruses 

Lung-JI, Chang, 1988 

Click here to the view this attachment (PDF). 

Special 


Transfection of this construct in mammalian cells produces a high concentration of 

VSV-G which can be used for pseudotyping retroviral- and lentiviral-based vectors. 

Note that VSV-G causes severe cell-cell fusion in 293 cell cultures. 

Recommended 

Storage: 

-20°C. 

Contributor: Dr. Lung-Ji Chang. 

References: 

Chang L-J, Urlacher V, Iwakuma T, Cui Y, Zucali J. Efficacy and safety analyses of a 

recombinant human immunodeficiency virus type 1 derived vector system. Gene Ther 

6:715?728, 1999. 

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pHEF-VSVG 

from Dr. Lung-Ji Chang." Also include the reference cited above in any publications. 




REV: 11/08/2015 Page 1 of 2




REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

pHEF-VSVG 




Provided: 5 μg plasmid DNA, 0.071 μg/μL, in TE. 

Description: 

This construct is a VSV-G envelope eukaryotic expression vector. Transfection of this 

construct in mammalian cells produces a high concentration of VSV-G which can be 

used for pseudotyping retroviral- and lentiviral-based vectors. 

Special 


PCR-amplified VSV-G sequence with a preferred eukaryotic translational initiation 

codon (-CCACC-ATG) driven by a strong human elongation factor alpha promoter. 


Propagate in DH5α, Top-10, HB101, or STBL2. 




Recommended 

Storage: 

-80°C. 


References: 



6:715?728, 1999. 




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NOTE: 







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DATA SHEET 

Reagent: 

pHEF-VSVG 




Provided: 5 μg plasmid DNA, 0.153 μg/μL, in TE. 

Description: 

This construct is a VSV-G envelope eukaryotic expression vector. Transfection of this 

construct in mammalian cells produces a high concentration of VSV-G which can be 

used for pseudotyping retroviral- and lentiviral-based vectors. 

Special 


PCR-amplified VSV-G sequence with a preferred eukaryotic translational initiation 

codon (-CCACC-ATG) driven by a strong human elongation factor alpha promoter. 


Propagate in DH5α, Top-10, HB101, or STBL2. 




Recommended 

Storage: 

-80°C. 


References: 



6:715?728, 1999. 




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NOTE: 







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XMRV >> Entry Clones

DATA SHEET 

Reagent: 

8400-E03 (eSD-H6-CA) 




Provided: 

Purified plasmid DNA (240 ng/µL) in TE. Can be propagated in any E. coli host strain 

using 50 µg/mL spectinomycin for selection. 

Description: XMRV (CA) gene cloned with a 5' optimized E.coli ribosome binding site (T7 gene 10 

enhancer plus Shine-Delgarno sequence), and an aminoterminal His6 non-cleavable 

fusion tag. Can be used to generate expression vectors for His6-fusions of XMRV 

proteins in E.coli. 

Cloning Site: Cloned using Gateway BP recombinational cloning. 


pDonr253 (modified version of Invitrogen pDonr201 vector with spectinomycin 

resistance instead of kanamycin resistance). 

Special 


Sequence File 

Table of Available XMRV Entry and Expression Clones 

Recommended 

Storage: 

-20°C or -80°C for long-term storage. 

Contributor: 

Drs. Stuart LeGrice and Alan Rein (NCI-Frederick) 

Drs. James Hartley, Dominic Esposito, Bill Gillette, Ralph Hopkins and Troy Taylor 

(SAIC-Frederick, Inc) 




REV: 11/08/2015 Page 1 of 2

NOTE: 


through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: (specify clone)." 


reagent must contact: Dr. Jeffrey W. Thomas, NCI Technology Transfer Center, 

ATRF Room E3202, PO Box B, Frederick, MD 21701, Email: 

jeffreyt@mail.nih.gov, Tel: (301) 846-5465, Fax: (301) 846-6820, before the 





REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E04 (eSD-H6-NC) 




Provided: 



Description: XMRV (NC) gene cloned with a 5′ optimized E.coli ribosome binding site (T7 gene 10 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











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DATA SHEET 

Reagent: 

8400-E05 (eSD-H6-PR) 




Provided: 



Description: XMRV (PR) gene cloned with a 5' optimized E.coli ribosome binding site (T7 gene 10 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











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DATA SHEET 

Reagent: 

8400-E06 (eSD-H6-RT) 




Provided: 



Description: XMRV (RT) gene cloned with a 5' optimized E.coli ribosome binding site (T7 gene 10 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 

Drs. Stuart LeGrice and Alan Rein (NCI-Frederick), and Drs. Dominic Esposito, William 

K. Gillette, Troy E. Taylor, Ralph F. Hopkins, and James L. Hartley (SAIC-Frederick, Inc) 




REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E07 (eSD-H6-IN) 




Provided: 



Description: XMRV (IN) gene cloned with a 5′ optimized E.coli ribosome binding site (T7 gene 10 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 






REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E08 (eSD-H6-SU) 




Provided: 



Description: XMRV (SU) gene cloned with a 5' optimized E.coli ribosome binding site (T7 gene 10 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 






REV: 11/08/2015 Page 1 of 2

NOTE: 











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DATA SHEET 

Reagent: 

8400-E09 (eSD-H6-TM) 




Provided: 



Description: XMRV (TM) gene cloned with a 5' optimized E.coli ribosome binding site (T7 gene 10 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 






REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E10 (eSD-H6-Env) 




Provided: 



Description: XMRV (Env) gene cloned with a 5' optimized E.coli ribosome binding site (T7 gene 10 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 






REV: 11/08/2015 Page 1 of 2

NOTE: 











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DATA SHEET 

Reagent: 

8400-E11 (tev-MA) 




Provided: 



Description: 

XMRV gene cloned with a 5′ tobacco etch virus (TEV) protease cleavage site 

(ENLYFQG), followed by a Kozak translation initiation sequence (ACC) and an ATG start 

codon. Can be used to generate expression vectors for aminoterminal fusions of XMRV 

proteins in any host system, which contain cleavable aminoterminal fusions. They can 

also be used to make native protein in eukaryotic hosts. 





Special 


Sequence File 


Recommended 

Storage: 


Contributor: Drs. Stuart LeGrice and Alan Rein (NCI-Frederick), and Drs. Dominic Esposito, William K. 

Gillette, Troy E. Taylor, Ralph F. Hopkins, and James L. Hartley (SAIC-Frederick, Inc) 




REV: 11/08/2015 Page 1 of 2

NOTE: 











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DATA SHEET 

Reagent: 

8400-E12 (tev-p12) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







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NOTE: 











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DATA SHEET 

Reagent: 

8400-E14 (tev-NC) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







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NOTE: 











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DATA SHEET 

Reagent: 

8400-E15 (tev-PR) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







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NOTE: 











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DATA SHEET 

Reagent: 

8400-E16 (tev-RT) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







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NOTE: 











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DATA SHEET 

Reagent: 

8400-E17 (tev-IN) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







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NOTE: 











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DATA SHEET 

Reagent: 

8400-E18 (tev-SU) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







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NOTE: 











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DATA SHEET 

Reagent: 

8400-E19 (tev-TM) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







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NOTE: 











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DATA SHEET 

Reagent: 

8400-E20 (tev-Env) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E22 (tev-p12-nostop) 




Provided: 



Description: 

XMRV gene cloned with a 5′ tobacco etch virus (TEV) protease cleavage site (ENLYFQG), 

followed by a Kozak translation initiation sequence (ACC) and an ATG start codon. 

These clones do NOT contain a stop codon at the 3’ end, and thus can be used to 

generate expression vectors for carboxyterminal fusions of XMRV proteins in any host 

system. They can also be used to make dual-tagged fusion proteins. 





Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E23 (tev-CA-nonstop) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E24 (tev-NC-nostop) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E25 (tev-PR-nostop) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E26 (tev-RT-nostop) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E27 (tev-IN-nostop) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E28 (tev-SU-nostop) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E29 (tev-TM-nostop) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E30 (tev-Env-nostop) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E01 (eSD-H6-MA) 




Provided: 



Description: XMRV (MA) gene cloned with a 5' optimized E.coli ribosome binding site (T7 gene 10 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E02 (eSD-H6-p12) 




Provided: 



Description: XMRV (p12) gene cloned with a 5' optimized E.coli ribosome binding site (T7 gene 10 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E13 tev-CA 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E21 (tev-MA-nostop) 




Provided: 



Description: 










Special 


Sequence File 


Recommended 

Storage: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E31 (SU-His6) 




Provided: 



Description: 

XMRV genes cloned with the native SU/Env signal peptide, and a noncleavable 

C-terminal His6 fusion tag. These clones can be used to generate expression clones for 

secretion of C-terminal His6 fusions of SU or Env from eukaryotic hosts. 





Special 


Sequence File 


Recommended 

Storage: 


Contributor: 



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REV: 11/08/2015 Page 1 of 2






REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-E33 (Env-His6) 




Provided: 



Description: 

XMRV genes cloned with the native SU/Env signal peptide, and a noncleavable 

C-terminal His6 fusion tag. These clones can be used to generate expression clones for 

secretion of C-terminal His6 fusions of SU or Env from eukaryotic hosts. 





Special 


Sequence File 


Recommended 

Storage: 


Contributor: 



NOTE: 










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XMRV >> Expression Vectors

DATA SHEET 

Reagent: 

8400-X31-008 (SU-His6) 




Provided: 


using 100 ug/ml ampicillin for selection. Requires transformation into a specialized 

bacmid production strain (E. coli DH10Bac) for generating of baculovirus. 

Description: 

XMRV genes designed to secrete XMRV Env or SU with a non-cleavable C-terminal His6 

tag. These vectors can be used to generate baculovirus by introduction into the 

DH10Bac strain (Invitrogen) and subsequent in vivo transposition (see Invitrogen 

Bac-to-Bac manual for more information). 

Cloning Site: Subcloned using Gateway LR recombinational cloning. 

Cloning Vector: pDest-8 (Baculovirus transfer vector from Invitrogen). 

Special 


Sequence File 


Recommended 

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DATA SHEET 

Reagent: 

8400-X33-008 (Env-His6) 




Provided: 



bacmid production strain (E. coli DH10Bac) for generating of baculovirus. 

Description: 

XMRV genes designed to secrete XMRV Env or SU with a non-cleavable C-terminal His6 

tag. These vectors can be used to generate baculovirus by introduction into the 

DH10Bac strain (Invitrogen) and subsequent in vivo transposition (see Invitrogen 

Bac-to-Bac manual for more information). 


Cloning Vector: pDest-8 (Baculovirus transfer vector from Invitrogen). 

Special 


Sequence File 


Recommended 

Storage: 


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DATA SHEET 

Reagent: 

8400-X01-521 (eSD-H6-MA) 




Provided: 


using 100 ug/ml ampicillin for selection. Requires a host strain which contains the T7 

RNA Polymerase gene (such as BL21(DE3)) for expression in E. coli. 

Description: 

XMRV genes with an aminoterminal non-cleavable His6 fusion tag. Protein expression is 

controlled by the T7 promoter and regulated by the presence of the lacI gene on the 

vector. Expression requires transformation into a strain which is capable of producing 

T7 RNA polymerase, and induction using IPTG. 


Cloning Vector: pDest-521 (based on Novagen’s pET43 series of vectors). 

Special 


Sequence File 


Recommended 

Storage: 


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REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X11-566 (His6-MBP-tev-MA) 




Provided: 




Description: 

XMRV genes with an aminoterminal cleavable His6-MBP (maltose binding protein) 

solubility tag. Protein expression is controlled by the T7 promoter and regulated by the 

presence of the lacI gene on the vector. Expression requires transformation into a strain 

which is capable of producing T7 RNA polymerase, and induction using IPTG. Tag can be 

removed by cleavage with tobacco etch virus (TEV) protease. 



pDest-566 (Gateway modified T7 vector containing MBP region from NEB pMal2 series 

vectors). 

Special 


Sequence File 


Recommended 

Storage: 


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REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X11-606 (H6-MBP-tev-MA) 




Provided: 



bacmid production strain (E. coli DH10Bac) for generating of baculovirus 

Description: 


solubility tag. Tag can be removed by cleavage with tobacco etch virus (TEV) protease. 

These vectors can be used to generate baculovirus by introduction into the DH10Bac 

strain (Invitrogen) and subsequent in vivo transposition (see Invitrogen Bac-to-Bac 

manual for more information). 


Cloning Vector: pDest-606 (Baculovirus transfer vector with His6-MBP aminoterminal fusion tag). 

Special 


Sequence File 


Recommended 

Storage: 


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REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X02-521 (eSD-H6-p12) 




Provided: 




Description: 







Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X03-521 (eSD-H6-CA) 




Provided: 




Description: 







Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X04-521 (eSD-H6-NC) 




Provided: 




Description: 







Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X05-521 (eSD-H6-PR) 




Provided: 




Description: 







Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X06-521 (eSD-H6-RT) 




Provided: 




Description: 







Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X07-521 (eSD-H6-IN) 




Provided: 




Description: 







Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X08-521 (eSD-H6-SU) 




Provided: 




Description: 







Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X09-521 (eSD-H6-TM) 




Provided: 




Description: 







Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X10-521 (eSD-H6-Env) 




Provided: 




Description: 







Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X12-566 (His6-MBP-tev-p12) 




Provided: 




Description: 









vectors). 

Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X13-566 (His6-MBP-tev-CA) 




Provided: 




Description: 









vectors). 

Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X14-566 (His6-MBP-tev-NC) 




Provided: 




Description: 









vectors). 

Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X15-566 (His6-MBP-tev-PR) 




Provided: 




Description: 









vectors). 

Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X17-566 (His6-MBP-tev-IN) 




Provided: 




Description: 









vectors). 

Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X16-566 (His6-MBP-tev-RT 




Provided: 




Description: 









vectors). 

Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X18-566 (His6-MBP-tev-SU) 




Provided: 




Description: 









vectors). 

Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X19-566 (His6-MBP-tev-TM) 




Provided: 




Description: 









vectors). 

Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X20-566 (His6-MBP-tev-Env) 




Provided: 




Description: 









vectors). 

Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X12-606 (H6-MBP-tev-p12) 




Provided: 




Description: 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X13-606 (H6-MBP-tev-CA) 




Provided: 




Description: 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X14-606 (H6-MBP-tev-NC) 




Provided: 




Description: 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X15-606 (H6-MBP-tev-PR) 




Provided: 




Description: 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X16-606 (H6-MBP-tev-RT) 




Provided: 




Description: 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X17-606 (H6-MBP-tev-IN) 




Provided: 




Description: 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X18-606 (H6-MBP-tev-SU) 




Provided: 




Description: 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X19-606 (H6-MBP-tev-TM) 




Provided: 




Description: 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







REV: 11/08/2015 Page 1 of 2

NOTE: 











REV: 11/08/2015 Page 2 of 2

DATA SHEET 

Reagent: 

8400-X20-606 (H6-MBP-tev-Env) 




Provided: 




Description: 








Special 


Sequence File 


Recommended 

Storage: 


Contributor: 







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NOTE: 











REV: 11/08/2015 Page 2 of 2

Expression Vectors

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