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pCLIP-Cas9-Nuclease-EFS-Puro

Lentiviral CRISPR vector encoding Cas9 and a puromycin resistance marker expressed from the EF-1α core promoter.

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pCLIP-Cas9-Nuclease-EFS-Puro.dna
Map and Sequence File:    Download    Open   
Sequence Author:  transOMIC
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SspI (10,575) PvuI (10,141) FspI (9993) DrdI (8989) AflIII - PciI (8881) SmaI (8861) TspMI - XmaI (8859) AvrII (8838) PmeI (8513) PsiI (8463) KpnI (8128) Acc65I (8124) Bsu36I (8120) PaeR7I - XhoI (8006) SalI (8000) BspDI - ClaI (7992) EcoRI (7378) BstEII (6912) RsrII (6894) BsiWI (6834) PflFI - Tth111I (6820) BamHI (6715) BsaBI * (6037) SpeI (41) NdeI (276) SnaBI (382) NruI * (867) BspQI - SapI (1136) NotI (1178) BbvCI (1454) AleI (1607) AgeI (2516) XbaI (2522) AfeI (2529) PspOMI (3894) ApaI (3898) PmlI (5328) pCLIP-Cas9-Nuclease-EFS-Puro 10,693 bp
SspI  (10,575)
1 site
A A T A T T T T A T A A
PvuI  (10,141)
1 site
C G A T C G G C T A G C
FspI  (9993)
1 site
T G C G C A A C G C G T
DrdI  (8989)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
AflIII  (8881)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (8881)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SmaI  (8861)
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  (8859)
1 site
C C C G G G G G G C C C
XmaI  (8859)
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.
AvrII  (8838)
1 site
C C T A G G G G A T C C
PmeI  (8513)
1 site
G T T T A A A C C A A A T T T G
PsiI  (8463)
1 site
T T A T A A A A T A T T
KpnI  (8128)
1 site
G G T A C C C C A T G G
Acc65I  (8124)
1 site
G G T A C C C C A T G G
Bsu36I  (8120)
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.
PaeR7I  (8006)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (8006)
1 site
C T C G A G G A G C T C
SalI  (8000)
1 site
G T C G A C C A G C T G
BspDI  (7992)
1 site
A T C G A T T A G C T A
ClaI  (7992)
1 site
A T C G A T T A G C T A
EcoRI  (7378)
1 site
G A A T T C C T T A A G
BstEII  (6912)
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.
RsrII  (6894)
1 site
C G G W C C G G C C W G