pGADT7 AD

Yeast two-hybrid "prey" vector for expressing proteins fused to the GAL4 activation domain.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsYeast PlasmidspGADT7 AD
SbfI (12) NsiI (7698) BsaAI - SnaBI (7342) BmgBI (6900) AatII (6568) ZraI (6566) PvuI (6016) NmeAIII (5794) BmrI (5686) AhdI (5646) AlwNI (5169) BspQI - SapI (4637) BspEI * (4526) NotI (4517) BmtI (4499) NheI (4495) PpuMI (3919) KasI (144) NarI (145) SfoI (146) PluTI (148) NgoMIV (210) NaeI (212) Bpu10I (291) BsmI (325) BtgZI (351) PacI (685) BsaBI (1009) ADH1 promoter BsgI (1421) ATG SV40 NLS Acc65I (1548) KpnI (1552) RsrII (1626) CsiI - SexAI * (1766) MluI (1795) BglII (1897) T7 promoter ATG HA NdeI (1969) SfiI (1980) EcoRI (1989) TspMI - XmaI (1997) AleI - SmaI (1999) BspDI - ClaI (2009) BamHI (2017) Eco53kI (2028) PaeR7I - PspXI - XhoI (2029) BanII - SacI (2030) AflII (3494) BstEII (3774) PflMI (3903) pGADT7 AD 7987 bp
SbfI  (12)
1 site		 C C T G C A G G G G A C G T C C
NsiI  (7698)
1 site		 A T G C A T T A C G T A
BsaAI  (7342)
1 site		 Y A C G T R R T G C A Y
SnaBI  (7342)
1 site		 T A C G T A A T G C A T
BmgBI  (6900)
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.
AatII  (6568)
1 site		 G A C G T C C T G C A G
ZraI  (6566)
1 site		 G A C G T C C T G C A G
PvuI  (6016)
1 site		 C G A T C G G C T A G C
NmeAIII  (5794)
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).
BmrI  (5686)
1 site		 A C T G G G ( N ) 4 N T G A C C C ( N ) 4
The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
AhdI  (5646)
1 site		 G A C N N N N N G T C C T G N N N N N C A G
The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
AlwNI  (5169)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
BspQI  (4637)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different BspQI sites may not be compatible.
SapI  (4637)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
BspEI  (4526)
1 site		 T C C G G A A G G C C T
* Blocked by Dam methylation.
NotI  (4517)
1 site		 G C G G C C G C C G C C G G C G
BmtI  (4499)
1 site		 G C T A G C C G A T C G
NheI  (4495)
1 site		 G C T A G C C G A T C G
PpuMI  (3919)
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.
KasI  (144)
1 site		 G G C G C C C C G C G G
NarI  (145)
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.
SfoI  (146)
1 site		 G G C G C C C C G C G G
PluTI  (148)
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.
NgoMIV  (210)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (212)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
Bpu10I  (291)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BsmI  (325)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
BtgZI  (351)
1 site		 G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4
Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
PacI  (685)
1 site		 T T A A T T A A A A T T A A T T
BsaBI  (1009)
1 site		 G A T N N N N A T C C T A N N N N T A G
BsgI  (1421)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
Acc65I  (1548)
1 site		 G G T A C C C C A T G G
KpnI  (1552)
1 site		 G G T A C C C C A T G G
RsrII  (1626)
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.
CsiI  (1766)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (1766)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
MluI  (1795)
1 site		 A C G C G T T G C G C A
BglII  (1897)
1 site		 A G A T C T T C T A G A
NdeI  (1969)
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.
SfiI  (1980)
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  (1989)
1 site		 G A A T T C C T T A A G
TspMI  (1997)
1 site		 C C C G G G G G G C C C
XmaI  (1997)
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.
AleI  (1999)
1 site		 C A C N N N N G T G G T G N N N N C A C
SmaI  (1999)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BspDI  (2009)
1 site		 A T C G A T T A G C T A
ClaI  (2009)
1 site		 A T C G A T T A G C T A
BamHI  (2017)
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.
Eco53kI  (2028)
1 site		 G A G C T C C T C G A G
PaeR7I  (2029)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2029)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (2029)
1 site		 C T C G A G G A G C T C
BanII  (2030)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
SacI  (2030)
1 site		 G A G C T C C T C G A G
AflII  (3494)
1 site		 C T T A A G G A A T T C
BstEII  (3774)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
PflMI  (3903)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
2μ ori
6820 .. 7984  =  1165 bp
yeast 2μ plasmid origin of replication
2μ ori
6820 .. 7984  =  1165 bp
yeast 2μ plasmid origin of replication
LEU2
2719 .. 3813  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
LEU2
2719 .. 3813  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
AmpR
5573 .. 6433  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5573 .. 6364  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5573 .. 6433  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6365 .. 6433  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5573 .. 6433  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ADH1 promoter
771 .. 1475  =  705 bp
promoter for alcohol dehydrogenase 1
ADH1 promoter
771 .. 1475  =  705 bp
promoter for alcohol dehydrogenase 1
ori
4814 .. 5402  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
4814 .. 5402  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ATG
1491 .. 1493  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for expression in yeast
ATG
1491 .. 1493  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for expression in yeast
SV40 NLS
1521 .. 1541  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1521 .. 1541  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
GAL4 activation domain
1557 .. 1898  =  342 bp
114 amino acids  =  12.4 kDa
Product: activation domain of the GAL4 transcriptional activator
GAL4 activation domain
1557 .. 1898  =  342 bp
114 amino acids  =  12.4 kDa
Product: activation domain of the GAL4 transcriptional activator
ATG
1935 .. 1937  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for in vitro transcription/translation
ATG
1935 .. 1937  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for in vitro transcription/translation
HA
1941 .. 1967  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
1941 .. 1967  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
LEU2 promoter
3814 .. 4219  =  406 bp
LEU2 promoter
3814 .. 4219  =  406 bp
ADH1 terminator
2415 .. 2602  =  188 bp
transcription terminator for alcohol dehydrogenase 1
ADH1 terminator
2415 .. 2602  =  188 bp
transcription terminator for alcohol dehydrogenase 1
AmpR promoter
6434 .. 6538  =  105 bp
AmpR promoter
6434 .. 6538  =  105 bp
MCS
1968 .. 2035  =  68 bp
multiple cloning site
MCS
1968 .. 2035  =  68 bp
multiple cloning site
T7 promoter
1904 .. 1922  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1904 .. 1922  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  1714 .. 2058  =  345 bp
ORF:  114 amino acids  =  13.5 kDa
ORF:  68 .. 409  =  342 bp
ORF:  113 amino acids  =  12.8 kDa
ORF:  2840 .. 3151  =  312 bp
ORF:  103 amino acids  =  11.1 kDa
ORF:  6917 .. 7183  =  267 bp
ORF:  88 amino acids  =  10.3 kDa
ORF:  1491 .. 2051  =  561 bp
ORF:  186 amino acids  =  20.5 kDa
ORF:  5703 .. 5969  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5573 .. 6433  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  7832 .. 69  =  225 bp
ORF:  74 amino acids  =  8.6 kDa
ORF:  2719 .. 3813  =  1095 bp
ORF:  364 amino acids  =  39.0 kDa
ORF:  7659 .. 7949  =  291 bp
ORF:  96 amino acids  =  11.5 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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Luciferase Vectors
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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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