Invitrogen Bac to Bac Expression System

Cloning into pFastBac HT A, B, and C, continued
Multiple Cloning
Site of
pFastBac HT B
3901
3951
Below is the multiple cloning site for pFastBac HT B. The initiation ATG is
indicated in bold. Restriction sites are labeled to indicate the actual cleavage site.
The boxed nucleotide indicates the variable region. The complete sequence of
pFastBac HT B is available for downloading from our Web site
(www.invitrogen.com) or by contacting Technical Service (see page 62). For a map
and a description of the features of pFastBac HT, refer to the Appendix, pages 55-
56.
Polyhedrin promoter
Start of
Transcription
TAGATCATGG AGATAATTAA AATGATAACC ATCTCGCAAA TAAATAAGTA
wild-type ATG mutated to ATT
TTTTACTGTT TTCGTAACAG TTTTGTAATA AAAAAACCTA TAAATATTCC
4001
4050
4092
4134
4176
4218
GGATTATTCA TACCGTCCCA CCATCGGGCG CGGATCTCGG TCCGAAACC
6xHis tag
ATG TCG TAC TAC CAT CAC CAT CAC CAT CAC GAT TAC GAT ATC
Met Ser Tyr Tyr His His His His His His Asp Tyr Asp Ile
TEV recognition site Ehe I Nco I Bam H I
CCA ACG ACC GAA AAC CTG TAT TTT CAG GGC GCC ATG GGA TCC
Pro Thr Thr Glu Asn Leu Tyr Phe Gln Gly Ala Met Gly Ser
TEV cleavage site
EcoR I
Stu I Sal I Sst I Spe I Not I
GGA ATT CAA AGG CCT ACG TCG ACG AGC TCA CTA GTC GCG GCC
Gly Ile Glu Arg Pro Thr Ser Thr Ser Ser Leu Val Ala Ala
Nsp V Xba I Pst I Xho I
Sph I Kpn I
GCT TTC GAA TCT AGA GCC TGC AGT CTC GAG GCA TGC GGT ACC
Ala Phe Glu Ser Arg Ala Cys Ser Leu Glu Ala Cys Gly Thr
Hind III
SV40 polyadenylation signal
AAG CTT GTC GAG AAG TAC TAG AG GATCATAATC AGCCATACCA ...
Lys Leu Val Glu Lys Tyr ***
continued on next page
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