pGL3 Luciferase Reporter Vectors Technical Manual #TM033

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III.C. pGL3-Promoter VectorThe pGL3-Promoter Vector contains an SV40 promoter upstream of theluciferase gene. DNA fragments containing putative enhancer elements can beinserted either upstream or downstream of the promoter-luc+ transcriptionalunit.2202 SalI2196 BamHIHpaI 2094oriXbaI 1934Amp rpGL3-PromoterVector(5010bp)SV40 latepoly(A) signal(for luc+ reporter)f1 oriluc+Synthetic poly(A)signal / transcriptionalpause site(for backgroundreduction)KpnISacIMluINheISmaIXhoIBgIII5111521283236SV40 PromoterHindIII 245NcoI 2780748VA08_4AFigure 3. The pGL3-Promoter Vector circle map. Additional description: luc+,cDNA encoding the modified firefly luciferase; Amp r , gene conferring ampicillinresistance in E. coli; f1 ori, origin of replication derived from filamentous phage; ori,origin of plasmid replication in E. coli. Arrows within luc+ and the Amp r geneindicate the direction of transcription; the arrow in f1 ori indicates the direction ofssDNA strand synthesis.pGL3-Promoter Vector Sequence Reference Points:Enhancer(none)Multiple cloning region 1–41Promoter 48–250GLprimer2 binding region 281–303Luciferase gene (luc+) 280–1932SV40 late poly(A) signal 1964–2185RVprimer4 binding region 2253–2272ColE1-derived plasmid replication origin 2510β-lactamase gene (Amp r ) 3272–4132f1 origin 4264–4719Upstream poly(A) signal 4850–5003RVprimer3 binding region 4952–4971Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USAToll Free in USA 800-356-9526 · Phone 608-274-4330 · Fax 608-277-2516 · www.promega.comPrinted in USA.Part# TM033Revised 9/07 Page 5
III.D. pGL3-Control VectorThe pGL3-Control Vector contains SV40 promoter and enhancer sequences,resulting in strong expression of luc+ in many types of mammalian cells. Thisplasmid is useful in monitoring transfection efficiency, in general, and is aconvenient internal standard for promoter and enhancer activities expressedby pGL3 recombinants.Amp rSynthetic poly(A)signal / transcriptionalpause site(for backgroundreduction)2448 SalI2442 BamHIoripGL3-ControlVector(5256bp)f1 oriKpnISacIMluINheISmaIXhoIBgIII5111521283236SV40 PromoterSV40 EnhancerSV40 latepoly(A) signal(for luc+ reporter)HpaI 2094XbaI 1934luc+HindIII 245NcoI 278NarI 3130747VA08_4AFigure 4. pGL3-Control Vector circle map. Additional description: luc+, cDNAencoding the modified firefly luciferase; Amp r , gene conferring ampicillin resistancein E. coli; f1 ori, origin of replication derived from filamentous phage; ori, origin ofplasmid replication in E. coli. Arrows within luc+ and the Amp r gene indicate thedirection of transcription; the arrow in f1 ori indicates the direction of ssDNA strandsynthesis.pGL3-Control Vector Sequence Reference Points:Multiple cloning region 1–41Promoter 48–250Luciferase gene (luc+) 280–1932GLprimer2 binding site 281–303SV40 late poly(A) signal 1964–2185Enhancer 2205–2441RVprimer4 binding site 2499–2518ColE1-derived plasmid replication origin 2756β-lactamase gene (Amp r ) 3518–4378f1 origin 4510–4965upstream poly(A) signal 5096–5249RVprimer3 binding site 5198–5217Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USAToll Free in USA 800-356-9526 · Phone 608-274-4330 · Fax 608-277-2516 · www.promega.comPart# TM033Printed in USA.Page 6 Revised 9/07
Page 1 and 2: Technical ManualpGL3 Luciferase Rep
Page 3 and 4: I. DescriptionThe pGL3 Luciferase R
Page 5: III.B. pGL3-Enhancer VectorThe pGL3
Page 9 and 10: IV.B. Preparation of pGL3 Vectors a
Page 11 and 12: VI.Assay of Luciferase Activity (co
Page 13 and 14: VIII. AppendixVIII.A. Common Struct
Page 15: VIII.C. The pGL3 Vectors luc+ GeneM
Page 18 and 19: VIII.F. References1. Sambrook, J. e
Page 21: VIII.G. pGL3-Basic Vector Restricti
Page 25 and 26: VIII.I. pGL3-Promoter Vector Restri
Page 27: VIII.J. pGL3-Control Vector Restric
Page 30: pGL4 Luciferase VectorsProduct Size

pGL3 Luciferase Reporter Vectors Technical Manual #TM033

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