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pCLIP-Cas9-Nuclease-EFS-Blast

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

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pCLIP-Cas9-Nuclease-EFS-Blast.dna
Map and Sequence File:    Download    Open   
Sequence Author:  transOMIC
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SspI (10,377) FspI (9795) AflIII - PciI (8683) SmaI (8663) TspMI - XmaI (8661) AvrII (8640) SexAI * (8407) PmeI (8315) PsiI (8265) KpnI (7930) Acc65I (7926) Bsu36I (7922) PaeR7I - XhoI (7808) SalI (7802) BspDI - ClaI (7794) SacII (7707) EcoRI (7180) BamHI (6715) PmlI (5328) SpeI (41) NdeI (276) SnaBI (382) KasI (671) NarI (672) SfoI (673) PluTI (675) BssHII (745) BspQI - SapI (1136) NotI (1178) BbvCI (1454) AleI (1607) AgeI (2516) XbaI (2522) AfeI (2529) PspOMI (3894) ApaI (3898) PasI (4308) pCLIP-Cas9-Nuclease-EFS-Blast 10,495 bp
SspI  (10,377)
1 site
A A T A T T T T A T A A
FspI  (9795)
1 site
T G C G C A A C G C G T
AflIII  (8683)
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  (8683)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SmaI  (8663)
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  (8661)
1 site
C C C G G G G G G C C C
XmaI  (8661)
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  (8640)
1 site
C C T A G G G G A T C C
SexAI  (8407)
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  (8315)
1 site
G T T T A A A C C A A A T T T G
PsiI  (8265)
1 site
T T A T A A A A T A T T
KpnI  (7930)
1 site
G G T A C C C C A T G G
Acc65I  (7926)
1 site
G G T A C C C C A T G G
Bsu36I  (7922)
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  (7808)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (7808)
1 site
C T C G A G G A G C T C
SalI  (7802)
1 site
G T C G A C C A G C T G
BspDI  (7794)
1 site
A T C G A T T A G C T A
ClaI  (7794)
1 site
A T C G A T T A G C T A
SacII  (7707)
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.
EcoRI  (7180)
1 site
G A A T T C C T T A A G
BamHI  (6715)
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.
PmlI  (5328)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
SpeI  (41)
1 site
A C T A G T T G A T C A
NdeI  (276)
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.
SnaBI  (382)
1 site
T A C G T A A T G C A T
KasI  (671)
1 site
G G C G C C C C G C G G
NarI  (672)
1 site
G G C G C C C C