Resources
Plasmid Files

M-ST1cas

Mammalian cell vector for expressing human codon-optimized Streptococcus thermophilus Cas9 with a G418-resistance marker.

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

M-ST1cas.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Church Lab / Addgene #48669
Download Free Trial Get SnapGene Viewer


MfeI (8789) SgrDI (8626) SspI (8509) PvuI (8075) AhdI (7705) PciI (6812) BstZ17I (6433) BsmI (6381) RsrII (5975) PflFI - Tth111I (5577) PluTI (5462) SfoI (5460) NarI (5459) KasI (5458) SmaI (5272) TspMI - XmaI (5270) AvrII (5249) PmeI (4181) MluI (43) SpeI (64) NdeI (299) SnaBI (405) Eco53kI (631) SacI (633) BsmBI (654) BspEI (716) XbaI (723) BstEII (813) SgrAI (929) SacII (1095) PspOMI (1689) ApaI (1693) AarI (1950) BsgI (1996) BstXI (2823) SbfI (2843) AscI (3642) FspAI (4045) AgeI (4163) M-ST1cas 8813 bp
MfeI  (8789)
1 site
C A A T T G G T T A A C
SgrDI  (8626)
1 site
C G T C G A C G G C A G C T G C
SspI  (8509)
1 site
A A T A T T T T A T A A
PvuI  (8075)
1 site
C G A T C G G C T A G C
AhdI  (7705)
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.
PciI  (6812)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (6433)
1 site
G T A T A C C A T A T G
BsmI  (6381)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
RsrII  (5975)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
PflFI  (5577)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (5577)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PluTI  (5462)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (5460)
1 site
G G C G C C C C G C G G
NarI  (5459)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (5458)
1 site
G G C G C C C C G C G G
SmaI  (5272)
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  (5270)
1 site
C C C G G G G G G C C C
XmaI  (5270)
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  (5249)
1 site
C C T A G G G G A T C C
PmeI  (4181)
1 site
G T T T A A A C C A A A T T T G
MluI  (43)
1 site
A C G C G T T G C G C A
SpeI  (64)
1 site
A C T A G T T G A T C A
NdeI  (299)
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  (405)
1 site
T A C G T A A T G C A T
Eco53kI  (631)
1 site
G A G C T C C T C G A G
SacI  (633)
1 site
G A G C T C C T C G A G
BsmBI  (654)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BspEI  (716)
1 site
T C C G G A A G G C C T
XbaI  (723)
1 site
T C T A G A A G A T C T
BstEII  (813)
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.
SgrAI  (929)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
SacII  (1095)
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.
PspOMI  (1689)
1 site
G G G C C C C C C G G G
ApaI  (1693)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AarI  (1950)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BsgI  (1996)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BstXI  (2823)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
SbfI  (2843)
1 site
C C T G C A G G G G A C G T C C
AscI  (3642)
1 site
G G C G C G C C C C G C G C G G
FspAI  (4045)
1 site
R T G C G C A Y Y A C G C G T R
AgeI  (4163)
1 site
A C C G G T T G G C C A
St1Cas9
748 .. 4110  =  3363 bp
1121 amino acids  =  129.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
St1Cas9
748 .. 4110  =  3363 bp
1121 amino acids  =  129.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus thermophilus Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
AmpR
7632 .. 8492