Resources
Plasmid Files

pGADT7 AD

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

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

pGADT7 AD Sequence and MappGADT7 AD.dna
Map and Sequence File   
Sequence Author:  Clontech
Download Free Trial Get SnapGene Viewer

 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) lac operator M13 rev 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) 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) PpuMI (3919) 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
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

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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

The sticky ends produced by AflII are hard to ligate.
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.
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.
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
   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
   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
LEU2 promoter
3814 .. 4219  =  406 bp
LEU2 promoter
3814 .. 4219  =  406 bp
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
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
lac promoter
4309 .. 4339  =  31 bp
   Segment 3:  -10  
   4309 .. 4315  =  7 bp
promoter for the E. coli lac operon
lac promoter
4309 .. 4339  =  31 bp
   Segment 2:  
   4316 .. 4333  =  18 bp
promoter for the E. coli lac operon
lac promoter
4309 .. 4339  =  31 bp
   Segment 1:  -35  
   4334 .. 4339  =  6 bp
promoter for the E. coli lac operon
lac promoter
4309 .. 4339  =  31 bp
3 segments
promoter for the E. coli lac operon
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
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
T7 promoter
1904 .. 1922  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1904 .. 1922  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
4261 .. 4277  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
4261 .. 4277  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
4285 .. 4301  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
4285 .. 4301  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
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
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
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter