pAD-GAL4

Phagemid vector carrying the GAL4 activation domain for yeast two-hybrid analysis.

Sequence Author: I.M.A.G.E. Consortium

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HomePlasmidsYeast PlasmidspAD-GAL4
SnaBI (6978) BmgBI (6536) PfoI (6315) AatII (6204) ZraI (6202) ScaI (5762) PvuI (5652) BsaI (5343) AhdI (5282) AlwNI (4805) BspQI - SapI (4273) EagI - NotI (4201) DraIII (3962) BtgZI (3954) NaeI (3856) BsgI (352) ATG Acc65I (479) KpnI (483) RsrII (557) MluI (726) EcoRI (833) SrfI (842) PaeR7I - PspXI - XhoI (847) SalI (858) XbaI (864) T7 promoter PstI (895) BglII (897) MscI (1033) BsrGI (1551) EcoRV (1926) AflII (2357) BspDI - ClaI (2522) BstEII (2637) PflMI (2766) PpuMI (2782) SfiI (3303) NcoI (3342) Bsu36I (3592) NgoMIV (3854) pAD-GAL4 7620 bp
SnaBI  (6978)
1 site		 T A C G T A A T G C A T
BmgBI  (6536)
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.
PfoI  (6315)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
AatII  (6204)
1 site		 G A C G T C C T G C A G
ZraI  (6202)
1 site		 G A C G T C C T G C A G
ScaI  (5762)
1 site		 A G T A C T T C A T G A
PvuI  (5652)
1 site		 C G A T C G G C T A G C
BsaI  (5343)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (5282)
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  (4805)
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  (4273)
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  (4273)
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.
EagI  (4201)
1 site		 C G G C C G G C C G G C
NotI  (4201)
1 site		 G C G G C C G C C G C C G G C G
DraIII  (3962)
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.
BtgZI  (3954)
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.
NaeI  (3856)
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.
BsgI  (352)
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  (479)
1 site		 G G T A C C C C A T G G
KpnI  (483)
1 site		 G G T A C C C C A T G G
RsrII  (557)
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.
MluI  (726)
1 site		 A C G C G T T G C G C A
EcoRI  (833)
1 site		 G A A T T C C T T A A G
SrfI  (842)
1 site		 G C C C G G G C C G G G C C C G
PaeR7I  (847)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (847)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (847)
1 site		 C T C G A G G A G C T C
SalI  (858)
1 site		 G T C G A C C A G C T G
XbaI  (864)
1 site		 T C T A G A A G A T C T
PstI  (895)
1 site		 C T G C A G G A C G T C
BglII  (897)
1 site		 A G A T C T T C T A G A
MscI  (1033)
1 site		 T G G C C A A C C G G T
BsrGI  (1551)
1 site		 T G T A C A A C A T G T
BsrGI is typically used at 37°C, but is even more active at 60°C.
EcoRV  (1926)
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.
AflII  (2357)
1 site		 C T T A A G G A A T T C
BspDI  (2522)
1 site		 A T C G A T T A G C T A
ClaI  (2522)
1 site		 A T C G A T T A G C T A
BstEII  (2637)
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  (2766)
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.
PpuMI  (2782)
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.
SfiI  (3303)
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.
NcoI  (3342)
1 site		 C C A T G G G G T A C C
Bsu36I  (3592)
1 site		 C C T N A G G G G A N T C C
Sticky ends from different Bsu36I sites may not be compatible.
NgoMIV  (3854)
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.
2μ ori
6456 .. 7620  =  1165 bp
yeast 2μ plasmid origin of replication
2μ ori
6456 .. 7620  =  1165 bp
yeast 2μ plasmid origin of replication
LEU2
1582 .. 2676  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
LEU2
1582 .. 2676  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
AmpR
5209 .. 6069  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5209 .. 6000  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5209 .. 6069  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6001 .. 6069  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5209 .. 6069  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
4450 .. 5038  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4450 .. 5038  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3729 .. 4184  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3729 .. 4184  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
ATG
422 .. 424  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
422 .. 424  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
452 .. 472  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
452 .. 472  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
GAL4 activation domain
488 .. 829  =  342 bp
114 amino acids  =  12.4 kDa
Product: activation domain of the GAL4 transcriptional activator
GAL4 activation domain
488 .. 829  =  342 bp
114 amino acids  =  12.4 kDa
Product: activation domain of the GAL4 transcriptional activator
LEU2 promoter
2677 .. 3081  =  405 bp
LEU2 promoter
2677 .. 3081  =  405 bp
ADH1 promoter
5 .. 406  =  402 bp
promoter for alcohol dehydrogenase 1
ADH1 promoter
5 .. 406  =  402 bp
promoter for alcohol dehydrogenase 1
ADH1 terminator
1277 .. 1464  =  188 bp
transcription terminator for alcohol dehydrogenase 1
ADH1 terminator
1277 .. 1464  =  188 bp
transcription terminator for alcohol dehydrogenase 1
AmpR promoter
6070 .. 6174  =  105 bp
AmpR promoter
6070 .. 6174  =  105 bp
MCS
833 .. 869  =  37 bp
multiple cloning site
MCS
833 .. 869  =  37 bp
multiple cloning site
T7 promoter
872 .. 890  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
872 .. 890  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  6553 .. 6819  =  267 bp
ORF:  88 amino acids  =  10.3 kDa
ORF:  422 .. 868  =  447 bp
ORF:  148 amino acids  =  16.1 kDa
ORF:  1703 .. 2014  =  312 bp
ORF:  103 amino acids  =  11.1 kDa
ORF:  5339 .. 5605  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  645 .. 878  =  234 bp
ORF:  77 amino acids  =  9.0 kDa
ORF:  1582 .. 2676  =  1095 bp
ORF:  364 amino acids  =  39.0 kDa
ORF:  5209 .. 6069  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  7295 .. 7585  =  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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