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pAC91-pmax-dCas9VP64

CRISPR activation vector for expressing catalytically inactive dCas9 fused to the VP64 transcriptional activation domain.

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pAC91-pmax-dCas9VP64.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Jaenisch Lab / Addgene #48223
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PluTI (7189) SfoI (7187) NarI (7186) KasI (7185) FspI (7086) PflFI - Tth111I (7069) RsrII (6669) ApaLI (6108) PciI (5794) StuI (5784) AflII (5749) HpaI (5657) PsiI (5637) Bts α I (5583) MluI (5519) BstBI (5475) SacII (5471) XbaI (5458) PaeR7I - PspXI - XhoI (5452) NotI (5446) PacI (5392) BspDI - ClaI (5380) AscI - BssHII (5219) BamHI (5211) FseI (5208) KflI (4442) BspHI (4189) DraIII (3464) NsiI (7371) NdeI (387) SnaBI (493) SgrAI (993) BglII (1315) PasI (2834) AhdI (3083) Eco53kI (3302) SacI (3304) pAC91-pmax-dCas9VP64 7373 bp
PluTI  (7189)
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  (7187)
1 site
G G C G C C C C G C G G
NarI  (7186)
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  (7185)
1 site
G G C G C C C C G C G G
FspI  (7086)
1 site
T G C G C A A C G C G T
PflFI  (7069)
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  (7069)
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.
RsrII  (6669)
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.
ApaLI  (6108)
1 site
G T G C A C C A C G T G
PciI  (5794)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
StuI  (5784)
1 site
A G G C C T T C C G G A
AflII  (5749)
1 site
C T T A A G G A A T T C
HpaI  (5657)
1 site
G T T A A C C A A T T G
PsiI  (5637)
1 site
T T A T A A A A T A T T
BtsαI  (5583)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
MluI  (5519)
1 site
A C G C G T T G C G C A
BstBI  (5475)
1 site
T T C G A A A A G C T T
SacII  (5471)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
XbaI  (5458)
1 site
T C T A G A A G A T C T
PaeR7I  (5452)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (5452)
1 site
V C T C G A G B B G A G C T C V
XhoI  (5452)
1 site
C T C G A G G A G C T C
NotI  (5446)
1 site
G C G G C C G C C G C C G G C G
PacI  (5392)
1 site
T T A A T T A A A A T T A A T T
BspDI  (5380)
1 site
A T C G A T T A G C T A
ClaI  (5380)
1 site
A T C G A T T A G C T A
AscI  (5219)
1 site
G G C G C G C C C C G C G C G G
BssHII  (5219)
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.
BamHI  (5211)
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.
FseI  (5208)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
KflI  (4442)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
BspHI  (4189)
1 site
T C A T G A A G T A C T
DraIII  (3464)
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.
NsiI  (7371)
1 site
A T G C A T T A C G