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pGADT7-Rec

High-copy yeast vector for fusing a cDNA to the GAL4 activation domain by in vivo recombination.

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pGADT7-Rec.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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SbfI (12) NsiI (7769) BsaAI - SnaBI (7413) BmgBI (6971) AatII (6639) ZraI (6637) PvuI (6087) BmrI (5757) AhdI (5717) AlwNI (5240) BspQI - SapI (4708) BspEI * (4597) NotI (4588) BmtI (4570) NheI (4566) KasI (144) NarI (145) SfoI (146) PluTI (148) NgoMIV (210) NaeI (212) Bpu10I (291) BsmI (325) BtgZI (351) PacI (685) BsaBI (1009) BsgI (1421) Acc65I (1548) KpnI (1552) RsrII (1626) SexAI * (1766) MluI (1795) BglII (1897) NdeI (1969) EcoRI (1989) TspMI - XmaI (2034) SmaI (2036) XbaI (2065) BspDI - ClaI (2080) BamHI (2088) Eco53kI (2099) PaeR7I - PspXI - XhoI (2100) BanII - SacI (2101) AflII (3565) BstEII (3845) PpuMI (3990) pGADT7-Rec 8058 bp
SbfI  (12)
1 site
C C T G C A G G G G A C G T C C
NsiI  (7769)
1 site
A T G C A T T A C G T A
BsaAI  (7413)
1 site
Y A C G T R R T G C A Y
SnaBI  (7413)
1 site
T A C G T A A T G C A T
BmgBI  (6971)
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  (6639)
1 site
G A C G T C C T G C A G
ZraI  (6637)
1 site
G A C G T C C T G C A G
PvuI  (6087)
1 site
C G A T C G G C T A G C
BmrI  (5757)
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  (5717)
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  (5240)
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  (4708)
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  (4708)
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  (4597)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
NotI  (4588)
1 site
G C G G C C G C C G C C G G C G
BmtI  (4570)
1 site
G C T A G C C G A T C G
NheI  (4566)
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.
EcoRI  (1989)
1 site
G A A T T C C T T A A G
TspMI  (2034)
1 site
C C C G G G G G G C C C
XmaI  (2034)
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  (2036)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
XbaI  (2065)
1 site
T C T A G A A G A T C T
BspDI  (2080)
1 site
A T C G A T T A G C T A
ClaI  (2080)
1 site
A T C G A T T A G C T A
BamHI  (2088)
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  (2099)
1 site
G A G C T C C T C G A G
PaeR7I  (2100)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2100)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2100)
1 site
C T C G A G G A G C T C
BanII  (2101)
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  (2101)
1 site
G A G C T C C T C G A G
AflII  (3565)
1 site
C T T A A