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pLenti-Cas9-IRES-Puro

3rd-generation lentiviral vector for expressing Cas9 and a puromycin resistance marker under control of the CMV promoter.

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pLenti-Cas9-IRES-Puro.dna
Map and Sequence File:    Download    Open   
Sequence Author:  OriGene
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PasI (11,134) ScaI (10,952) Bsu36I (9622) BsrGI (8782) StuI (8513) SmaI (8233) TspMI - XmaI (8231) BmgBI (8150) AarI (7946) PmlI (7923) PmeI (7577) PaeR7I - PspXI - XhoI (7499) BsiWI (7487) MluI (7484) XcmI (6868) SgrDI (1) FseI (1530) EcoNI (1550) BbvCI (1804) PstI (2417) XbaI (2613) EcoRI (3279) BamHI (3292) AsiSI (3324) BstZ17I (3543) BlpI (4008) SphI (4589) PshAI (4904) pLenti-Cas9-IRES-Puro 11,716 bp
PasI  (11,134)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
ScaI  (10,952)
1 site
A G T A C T T C A T G A
Bsu36I  (9622)
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.
BsrGI  (8782)
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.
StuI  (8513)
1 site
A G G C C T T C C G G A
SmaI  (8233)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (8231)
1 site
C C C G G G G G G C C C
XmaI  (8231)
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.
BmgBI  (8150)
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.
AarI  (7946)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
PmlI  (7923)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
PmeI  (7577)
1 site
G T T T A A A C C A A A T T T G
PaeR7I  (7499)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (7499)
1 site
V C T C G A G B B G A G C T C V
XhoI  (7499)
1 site
C T C G A G G A G C T C
BsiWI  (7487)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
MluI  (7484)
1 site
A C G C G T T G C G C A
XcmI  (6868)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
SgrDI  (1)
1 site
C G T C G A C G G C A G C T G C
FseI  (1530)
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.
EcoNI  (1550)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BbvCI  (1804)
1 site
C C T C A G C G G A G T C G
PstI  (2417)
1 site
C T G C A G G A C G T C
XbaI  (2613)
1 site
T C T A G A A G A T C T
EcoRI  (3279)
1 site
G A A T T C C T T A A G
BamHI  (3292)
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.
AsiSI  (3324)
1 site
G C G A T C G C C G C T A G C G
BstZ17I  (3543)
1 site
G T A T A C C A T A T G
BlpI  (4008)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
SphI  (4589)
1 site
G C A T G C C G T A C G
PshAI  (4904)
1 site
G A C N N N N G T