pGBKT7

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

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsYeast PlasmidspGBKT7
DraIII (7031) PaqCI (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) T7 promoter ATG Myc 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.
PaqCI  (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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
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
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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
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
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
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
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
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
T7 promoter
1213 .. 1231  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1213 .. 1231  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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
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Individual Sequences & Maps

bleMX6
pFA6a-kanMX6
pPIC6(alpha) B
pYES2.1-E
hphMX6
pFA6a-kanMX6-PGAL1
pPIC6(alpha) C
pYES2 CT
kanMX
pFA6a-kanMX6-PGAL1-3HA
pPIC9
pYES2 NT A
kanMX2
pFA6a-kanMX6-PGAL1-GFP
pPIC9K
pYES2 NT B
kanMX4
pFA6a-kanMX6-PGAL1-GST
pPICZ A
pYES2 NT C
kanMX6
pFA6a-natMX6
pPICZ B
pYES3 CT
natMX6
pFA6a-TRP1
pPICZ C
pYES6 CT
pAbAi
pFA6a-TRP1-PGAL1
pPICZ(alpha) A
pYM25
pAD-GAL4
pFA6a-TRP1-PGAL1-3HA
pPICZ(alpha) B
pYM26
pAG25
pFA6a-TRP1-PGAL1-GFP
pPICZ(alpha) C
pYM44
pAG29
pFA6a-TRP1-PGAL1-GST
pPink-HC
YCp50
pAG32
pFLD
pPink-LC
YCp50-poly
pAO815
pFLD(alpha)
pPINK(alpha)-HC
YCplac111
patMX4
pGAD-C1
pRS303
YCplac22
pAUR101
pGAD-C2
pRS304
YCplac33
pAUR112
pGAD-C3
pRS305
YEp13
pAUR123
pGADT7 AD
pRS306
YEp24
pAUR224
pGADT7-Rec
pRS313
YEp351
pBridge
pGADT7-Rec (linearized)
pRS314
YEp352
pDH25
pGAPZ A
pRS315
YEp353
pEG202
pGAPZ B
pRS316
YEp354
pESC-HIS
pGAPZ C
pRS403
YEp355
pESC-LEU
pGAPZ(alpha) A
pRS404
YEp356
pESC-TRP
pGAPZ(alpha) B
pRS405
YEp356R
pESC-URA
pGAPZ(alpha) C
pRS406
YEp357
pFA6-kanMX4
pGBD-C1
pRS413
YEp357R
pFA6a
pGBD-C2
pRS414
YEp358
pFA6a-12Pk-kanMX6
pGBD-C3
pRS415
YEp358R
pFA6a-13Myc-His3MX6
pGBDU-C1
pRS416
YEp363
pFA6a-13Myc-kanMX6
pGBDU-C2
pRS423
YEp364
pFA6a-13Myc-TRP1
pGBDU-C3
pRS424
YEp365
pFA6a-3HA-His3MX6
pGBKT7
pRS425
YEp366
pFA6a-3HA-kanMX6
pGilda
pRS426
YEp366R
pFA6a-3HA-TRP1
pHIL-D2
pSpot5
YEp367
pFA6a-5FLAG-kanMX6
pHIL-S1
pSpot6
YEp367R
pFA6a-bleMX6
pJG4-5
pSpot7
YEp368
pFA6a-GFP(S65T)-His3MX6
pKLAC1
pSpot8
YEp368R
pFA6a-GFP(S65T)-kanMX6
pKLAC2
pTEF1 Zeo
YEplac112
pFA6a-GFP(S65T)-TRP1
pOAD
pUCATPH
YEplac181
pFA6a-GST-His3MX6
pOBD2
pY-secSUMOstar
YEplac195
pFA6a-GST-kanMX6
pP-secSUMO3
pY-SUMOstar
YIp351
pFA6a-GST-TRP1
pPIC3
pYC2-E
YIp352
pFA6a-His3MX6
pPIC3.5
pYC2 CT
YIp5
pFA6a-His3MX6-PGAL1
pPIC3.5K
pYC2 NT A
YIplac128
pFA6a-His3MX6-PGAL1-3HA
pPIC6 A
pYC2 NT B
YIplac204
pFA6a-His3MX6-PGAL1-GFP
pPIC6 B
pYC2 NT C
YIplac211
pFA6a-His3MX6-PGAL1-GST
pPIC6 C
pYC6 CT
YRp7
pFA6a-hphMX6
pPIC6(alpha) A
pYES2
alpha-factor secretion signal

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