pCMV6-AN-mYFP

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

Sequence Author: OriGene

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AanI (6562) ScaI (5765) PciI (4392) BstZ17I (4013) BsmI (3961) AanI (3900) BstBI (3723) PflFI - Tth111I (3159) BsrGI (301) NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) BmgBI (1085) BstEII (1202) PasI (1583) BspEI (1737) AsiSI - SgfI (1753) MreI - SgrAI (1755) AscI (1759) HindIII (1773) RsrII (1790) MluI (1796) NotI (1806) PaeR7I - PspXI - XhoI (1814) PmeI (1823) FseI (1833) XL39 (reverse primer) (1929 .. 1948) BbsI (2046) EcoO109I (2053) BbvCI (2149) BlpI (2193) BsmBI (2213) BstXI (2300) AgeI (2411) SexAI * (2598) SfiI (2784) StuI (2830) AvrII (2831) BclI * (2882) KasI (3040) NarI (3041) SfoI (3042) PluTI (3044) pCMV6-AN-mYFP 6634 bp
AanI  (6562)
2 sites
T T A T A A A A T A T T
ScaI  (5765)
1 site
A G T A C T T C A T G A
PciI  (4392)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (4013)
1 site
G T A T A C C A T A T G
BsmI  (3961)
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  (3900)
2 sites
T T A T A A A A T A T T
BstBI  (3723)
1 site
T T C G A A A A G C T T
PflFI  (3159)
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  (3159)
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.
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
BmgBI  (1085)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
BstEII  (1202)
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.
PasI  (1583)
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.
BspEI  (1737)
1 site
T C C G G A A G G C C T
AsiSI  (1753)
1 site
G C G A T C G C C G C T A G C G
SgfI  (1753)
1 site
G C G A T C G C C G C T A G C G
MreI  (1755)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (1755)
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  (1759)
1 site
G G C G C G C C C C G C G C G G
HindIII  (1773)
1 site
A A G C T T T T C G A A
RsrII  (1790)
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  (1796)
1 site
A C G C G T T G C G C A
NotI  (1806)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1814)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1814)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1814)
1 site
C T C G A G G A G C T C
PmeI  (1823)
1 site
G T T T A A A C C A A A T T T G
FseI  (1833)
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.
BbsI  (2046)
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.
EcoO109I  (2053)
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.
BbvCI  (2149)
1 site
C C T C A G C G G A G T C G
BlpI  (2193)
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  (2213)
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  (2300)
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  (2411)
1 site
A C C G G T T G G C C A
SexAI  (2598)
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  (2784)
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  (2830)
1 site
A