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pCLIP-Cas9-Nuclease-EFS-ZsGreen

Lentiviral CRISPR vector encoding Cas9 and the green fluorescent protein ZsGreen expressed from the EF-1α core promoter.

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pCLIP-Cas9-Nuclease-EFS-ZsGreen.dna
Map and Sequence File:    Download    Open   
Sequence Author:  transOMIC
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SspI (10,674) PvuI (10,240) FspI (10,092) DrdI (9088) AflIII - PciI (8980) SmaI (8960) TspMI - XmaI (8958) AvrII (8937) SexAI * (8704) PmeI (8612) PsiI (8562) KpnI (8227) Acc65I (8223) Bsu36I (8219) PaeR7I - XhoI (8105) BspDI - ClaI (8091) SacII (8004) EcoRI (7477) BsrGI (7331) PflFI - Tth111I (7015) SgrAI (6868) BamHI (6715) BsaBI * (6037) SpeI (41) NdeI (276) SnaBI (382) BssHII (745) NruI * (867) BspQI - SapI (1136) NotI (1178) BbvCI (1454) AleI (1607) AgeI (2516) XbaI (2522) AfeI (2529) PspOMI (3894) ApaI (3898) PasI (4308) PmlI (5328) pCLIP-Cas9-Nuclease-EFS-ZsGreen 10,792 bp
SspI  (10,674)
1 site
A A T A T T T T A T A A
PvuI  (10,240)
1 site
C G A T C G G C T A G C
FspI  (10,092)
1 site
T G C G C A A C G C G T
DrdI  (9088)
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  (8980)
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  (8980)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SmaI  (8960)
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  (8958)
1 site
C C C G G G G G G C C C
XmaI  (8958)
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  (8937)
1 site
C C T A G G G G A T C C
SexAI  (8704)
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.
PmeI  (8612)
1 site
G T T T A A A C C A A A T T T G
PsiI  (8562)
1 site
T T A T A A A A T A T T
KpnI  (8227)
1 site