Resources
Plasmid Files

pGBKT7

Yeast two-hybrid "bait" vector for expressing proteins fused to the GAL4 DNA-binding domain.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pGBKT7.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
Download Free Trial Get SnapGene Viewer


DraIII (7031) AarI (6462) BstXI (6327) EcoRV (6314) PmlI (5989) BspEI * (5894) XcmI (5437) BmgBI (5226) NsiI (4436) PluTI (3887) SfoI (3885) NarI (3884) KasI (3883) PflFI - Tth111I (3767) PvuI (0) BsaBI (268) PaeR7I - XhoI (979) HpaI (1039) PpuMI (1274) NdeI (1282) BtgI - NcoI (1288) SfiI (1293) EcoRI (1299) TspMI - XmaI (1304) SmaI (1306) BamHI (1309) SalI (1315) PstI (1325) NotI (1327) BssHII (1385) AvrII (3045) BstBI (3202) RsrII (3367) NmeAIII (3664) pGBKT7 7303 bp
DraIII  (7031)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
AarI  (6462)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BstXI  (6327)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
EcoRV  (6314)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
PmlI  (5989)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BspEI  (5894)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
XcmI  (5437)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BmgBI  (5226)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
NsiI  (4436)
1 site
A T G C A T T A C G T A
PluTI  (3887)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (3885)
1 site
G G C G C C C C G C G G
NarI  (3884)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (3883)
1 site
G G C G C C C C G C G G
PflFI  (3767)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3767)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PvuI  (0)
1 site
C G A T C G G C T A G C
BsaBI  (268)
1 site
G A T N N N N A T C C T A N N N N T A G
PaeR7I  (979)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (979)
1 site
C T C G A G G A G C T C
HpaI  (1039)
1 site
G T T A A C C A A T T G
PpuMI  (1274)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
NdeI  (1282)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BtgI  (1288)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1288)
1 site
C C A T G G G G T A C C
SfiI  (1293)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
EcoRI  (1299)
1 site
G A A T T C C T T A A G
TspMI  (1304)
1 site
C C C G G G G G G C C C
XmaI  (1304)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (1306)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (1309)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
SalI  (1315)
1 site
G T C G A C C A G C T G
PstI  (1325)
1 site
C T G C A G G A C G T C
NotI  (1327)
1 site
G C G G C C G C C G C C G G C G
BssHII  (1385)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
AvrII  (3045)
1 site
C C T A G G G G A T C C
BstBI  (3202)
1 site
T T C G A A A A G C T T
RsrII  (3367)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
NmeAIII  (3664)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
2μ ori
4147 .. 5489  =  1343 bp
yeast 2μ plasmid origin of replication
2μ ori
4147 .. 5489  =  1343 bp
yeast 2μ plasmid origin of replication
NeoR/KanR
3221 .. 4015  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
3221 .. 4015  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ADH1 promoter
30 .. 734  =  705 bp
promoter for alcohol dehydrogenase 1
ADH1 promoter
30 .. 734  =  705 bp
promoter for alcohol dehydrogenase 1
TRP1
6030 .. 6704  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
TRP1
6030 .. 6704  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
ori
2025 .. 2613  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
2025 .. 2613  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
f1 ori
6807 .. 7262  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6807 .. 7262  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
GAL4 DNA binding domain
762 .. 1202  =  441 bp
147 amino acids  =  16.9 kDa
Product: DNA binding domain of the GAL4 transcriptional activator
GAL4 DNA binding domain
762 .. 1202  =  441 bp
147 amino acids  =  16.9 kDa
Product: DNA binding domain of the GAL4 transcriptional activator
TRP1 promoter
5748 .. 6029  =  282 bp
TRP1 promoter
5748 .. 6029  =  282 bp
ADH1 terminator
1412 .. 1599  =  188 bp
transcription terminator for alcohol dehydrogenase 1
ADH1 terminator
1412 .. 1599  =  188 bp
transcription terminator for alcohol dehydrogenase 1
AmpR promoter
4016 .. 4120  =  105 bp
AmpR promoter
4016 .. 4120  =  105 bp
MCS
1281 .. 1333  =  53 bp
multiple cloning site
MCS
1281 .. 1333  =  53 bp
multiple cloning site
T7 terminator
1338 .. 1385  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1338 .. 1385  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
HSV TK poly(A) signal
2942 .. 2989  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
2942 .. 2989  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
Myc
1251 .. 1280  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
1251 .. 1280  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
T7 promoter
1213 .. 1231  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1213 .. 1231  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ATG
1245 .. 1247  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for in vitro transcription/translation
ATG
1245 .. 1247  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for in vitro transcription/translation
ORF:  6280 .. 6561  =  282 bp
ORF:  93 amino acids  =  10.8 kDa
ORF:  2762 .. 2995  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  3170 .. 3706  =  537 bp
ORF:  178 amino acids  =  19.9 kDa
ORF:  762 .. 1337  =  576 bp
ORF:  191 amino acids  =  21.8 kDa
ORF:  4182 .. 4472  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
ORF:  6030 .. 6704  =  675 bp
ORF:  224 amino acids  =  24.1 kDa
ORF:  3221 .. 4015  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  6440 .. 6802  =  363 bp
ORF:  120 amino acids  =  13.3 kDa
ORF:  3457 .. 3843  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  4948 .. 5268  =  321 bp
ORF:  106 amino acids  =  12.9 kDa
ORF:  2751 .. 3200  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps