pCMV6-AN-mKate

PrecisionShuttle™ mammalian expression vector for fusing an ORF to an N-terminal mKate2 tag.

Sequence Author: OriGene

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AanI (6553) XmnI (5875) ScaI (5756) PciI (4383) BstZ17I (4004) BsmI (3952) AanI (3891) BstBI (3714) CMV enhancer NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) PasI (1241) PshAI (1650) BspEI (1728) AsiSI - SgfI (1744) MreI - SgrAI (1746) AscI (1750) HindIII (1764) RsrII (1781) MluI (1787) NotI (1797) PaeR7I - PspXI - XhoI (1805) PmeI (1814) FseI (1824) SacII (1828) XL39 (reverse primer) (1920 .. 1939) EcoO109I (2044) BsgI (2098) BbvCI - Bpu10I (2140) BlpI (2184) BsmBI (2204) BstXI (2291) AgeI (2402) PflMI (2405) XcmI (2497) SexAI * (2589) SfiI (2775) AvrII (2822) BclI * (2873) BsaBI * (2891) KasI (3031) NarI (3032) SfoI (3033) PluTI (3035) PstI (3085) PflFI - Tth111I (3150) pCMV6-AN-mKate 6625 bp
AanI  (6553)
2 sites
T T A T A A A A T A T T
XmnI  (5875)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5756)
1 site
A G T A C T T C A T G A
PciI  (4383)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (4004)
1 site
G T A T A C C A T A T G
BsmI  (3952)
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.
AanI  (3891)
2 sites
T T A T A A A A T A T T
BstBI  (3714)
1 site
T T C G A A A A G C T T
NdeI  (592)
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  (698)
1 site
T A C G T A A T G C A T
Eco53kI  (924)
1 site
G A G C T C C T C G A G
SacI  (926)
1 site
G A G C T C C T C G A G
EcoRI  (979)
1 site
G A A T T C C T T A A G
BamHI  (992)
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.
Acc65I  (998)
1 site
G G T A C C C C A T G G
KpnI  (1002)
1 site
G G T A C C C C A T G G
PasI  (1241)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PshAI  (1650)
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.
BspEI  (1728)
1 site
T C C G G A A G G C C T
AsiSI  (1744)
1 site
G C G A T C G C C G C T A G C G
SgfI  (1744)
1 site
G C G A T C G C C G C T A G C G
MreI  (1746)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (1746)
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.
AscI  (1750)
1 site
G G C G C G C C C C G C G C G G
HindIII  (1764)
1 site
A A G C T T T T C G A A
RsrII  (1781)
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.
MluI  (1787)
1 site
A C G C G T T G C G C A
NotI  (1797)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1805)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1805)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1805)
1 site
C T C G A G G A G C T C
PmeI  (1814)
1 site
G T T T A A A C C A A A T T T G
FseI  (1824)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
SacII  (1828)
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.
EcoO109I  (2044)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
BsgI  (2098)
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).
BbvCI  (2140)
1 site
C C T C A G C G G A G T C G
Bpu10I  (2140)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BlpI  (2184)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BsmBI  (2204)
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.
BsmBI-v2 is an improved version of BsmBI.
BstXI  (2291)
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.
AgeI  (2402)
1 site
A C C G G T T G G C C A
PflMI  (2405)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
XcmI  (2497)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
SexAI  (2589)
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.
SfiI  (2775)
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.
AvrII  (2822)
1 site
C C T A G G G G A T C C
BclI  (2873)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BsaBI  (2891)
1 site
G A T