pCMV6-A-EM7-BSD

PrecisionShuttle™ mammalian vector for expressing an untagged ORF, with a blasticidin resistance marker that can also be used in bacteria.

Sequence Author: OriGene

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AanI (5500) DraIII (5375) XmnI (4822) ScaI (4703) AlwNI (3746) PciI (3330) BspQI - SapI (3214) BstZ17I (2951) BsmI (2899) AanI (2838) BsrGI (301) CMV enhancer NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) AsiSI - SgfI (1024) MreI - SgrAI (1026) AscI - BssHII (1030) HindIII (1044) RsrII (1061) MluI (1067) NotI (1077) PaeR7I - PspXI - XhoI (1085) PmeI (1094) FseI (1104) SacII (1108) AleI (1194) XL39 (reverse primer) (1200 .. 1219) BbvCI - Bpu10I (1420) BlpI (1464) BsmBI (1484) BstXI (1571) AgeI (1682) PflMI (1685) XcmI (1777) SexAI * (1869) SfiI (2055) StuI (2101) AvrII (2102) BclI * (2153) BsiWI (2166) PpuMI (2401) NruI * (2473) BsaBI * (2540) PstI (2645) BstBI (2661) BfuAI - BspMI (2697) PfoI (2754) pCMV6-A-EM7-BSD 5572 bp
AanI  (5500)
2 sites
T T A T A A A A T A T T
DraIII  (5375)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
XmnI  (4822)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (4703)
1 site
A G T A C T T C A T G A
AlwNI  (3746)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
PciI  (3330)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3214)
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  (3214)
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.
BstZ17I  (2951)
1 site
G T A T A C C A T A T G
BsmI  (2899)
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  (2838)
2 sites
T T A T A A A A T A T T
BsrGI  (301)
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.
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
AsiSI  (1024)
1 site
G C G A T C G C C G C T A G C G
SgfI  (1024)
1 site
G C G A T C G C C G C T A G C G
MreI  (1026)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (1026)
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  (1030)
1 site
G G C G C G C C C C G C G C G G
BssHII  (1030)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
HindIII  (1044)
1 site
A A G C T T T T C G A A
RsrII  (1061)
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  (1067)
1 site
A C G C G T T G C G C A
NotI  (1077)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1085)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1085)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1085)
1 site
C T C G A G G A G C T C
PmeI  (1094)
1 site
G T T T A A A C C A A A T T T G
FseI  (1104)
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  (1108)
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.
AleI  (1194)
1 site
C A C N N N N G T G G T G N N N N C A C
BbvCI  (1420)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1420)
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  (1464)
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  (1484)
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  (1571)
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  (1682)
1 site
A C C G G T T G G C C A
PflMI  (1685)
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  (1777)
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  (1869)
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  (2055)
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.
StuI  (2101)
1 site
A G G C C T T C C G G A
AvrII  (2102)
1 site
C C T A G G G G A T C C
BclI  (2153)
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.
BsiWI  (2166)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
PpuMI  (2401)
1 site
R G G W C