Resources
Plasmid Files

MSP469

Vector for expressing the S. pyogenes Cas9 VQR mutant, which has an altered PAM recognition specificity.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

MSP469.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Joung Lab / Addgene #65771
Download Free Trial Get SnapGene Viewer


SpeI (7380) MluI (7359) SgrDI (7125) ScaI (6684) PvuI (6574) BsaI (6265) AhdI (6204) BspQI - SapI (5195) BbsI (4864) PmeI (4653) AgeI (4623) PstI (4617) PsiI (4571) PaeR7I - XhoI (4539) BamHI (4513) BmtI (4055) NheI (4051) EagI - NotI - SacII (370) BstZ17I (623) BmgBI (628) MscI (679) Bsu36I (859) BstEII (1123) StuI (1148) BsiWI (1217) KasI (1350) NarI (1351) SfoI (1352) PluTI (1354) BfuAI - BspMI (1477) BsrGI (1967) PfoI * (2225) EcoRV (2250) PshAI (2391) FspAI (3359) BsgI (3727) MSP469 7614 bp
SpeI  (7380)
1 site
A C T A G T T G A T C A
MluI  (7359)
1 site
A C G C G T T G C G C A
SgrDI  (7125)
1 site
C G T C G A C G G C A G C T G C
ScaI  (6684)
1 site
A G T A C T T C A T G A
PvuI  (6574)
1 site
C G A T C G G C T A G C
BsaI  (6265)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (6204)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BspQI  (5195)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (5195)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
BbsI  (4864)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
PmeI  (4653)
1 site
G T T T A A A C C A A A T T T G
AgeI  (4623)
1 site
A C C G G T T G G C C A
PstI  (4617)
1 site
C T G C A G G A C G T C
PsiI  (4571)
1 site
T T A T A A A A T A T T
PaeR7I  (4539)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (4539)
1 site
C T C G A G G A G C T C
BamHI  (4513)
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.
BmtI  (4055)
1 site
G C T A G C C G A T C G
NheI  (4051)
1 site
G C T A G C C G A T C G
EagI  (370)
1 site
C G G C C G G C C G G C
NotI  (370)
1 site
G C G G C C G C C G C C G G C G
SacII  (370)
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.
BstZ17I  (623)
1 site
G T A T