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pCLIP-Cas9-Nuclease-hCMV-tRFP

Lentiviral CRISPR vector encoding Cas9 and the red fluorescent protein TurboRFP expressed from the human CMV promoter.

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pCLIP-Cas9-Nuclease-hCMV-tRFP.dna
Map and Sequence File:    Download    Open   
Sequence Author:  transOMIC
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SspI (11,040) PvuI (10,606) FspI (10,458) DrdI (9454) AflIII - PciI (9346) SmaI (9326) TspMI - XmaI (9324) AvrII (9303) SexAI * (9070) PmeI (8978) PsiI (8928) KpnI (8593) Acc65I (8589) SalI (8465) SacII (8370) PshAI (7774) SfiI (7610) BsrGI (7184) BsaBI * (6403) PmlI (5694) NheI (405) BmtI (409) KasI (671) NarI (672) SfoI (673) PluTI (675) BssHII (745) NruI * (867) BspQI - SapI (1136) MfeI (1219) BbvCI (1454) AleI (1607) BsmBI (2766) PspXI (2882) XbaI (2888) AfeI (2895) PspOMI (4260) ApaI (4264) pCLIP-Cas9-Nuclease-hCMV-tRFP 11,158 bp
SspI  (11,040)
1 site
A A T A T T T T A T A A
PvuI  (10,606)
1 site
C G A T C G G C T A G C
FspI  (10,458)
1 site
T G C G C A A C G C G T
DrdI  (9454)
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  (9346)
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  (9346)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SmaI  (9326)
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  (9324)
1 site
C C C G G G G G G C C C
XmaI  (9324)
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  (9303)
1 site
C C T A G G G G A T C C
SexAI  (9070)
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  (8978)
1 site
G T T T A A A C C A A A T T T G
PsiI  (8928)
1 site
T T A T A A A A T A T T
KpnI  (8593)
1 site
G G T A C C C C A T G G
Acc65I  (8589)
1 site
G G T A C C C C A T G G
SalI  (8465)
1 site
G T C G A C C A G C T G
SacII  (8370)
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.
PshAI  (7774)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
SfiI  (7610)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
BsrGI  (7184)
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.
BsaBI  (6403)
1 site
G A T N N N N A T C C T A N N N