pCMV6-AN-FP635

PrecisionShuttle™ mammalian expression vector for fusing an ORF to an N-terminal TurboFP635 (Katushka) tag.

Sequence Author: OriGene

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DraIII (6425) XmnI (5872) ScaI (5753) PciI (4380) BstZ17I (4001) BsmI (3949) AanI (3888) PfoI (3804) BstBI (3711) PflFI - Tth111I (3147) AanI (6550) NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) PasI (1250) Bsu36I (1285) PshAI (1659) BspEI (1725) AsiSI - SgfI (1741) MreI - SgrAI (1743) AscI (1747) HindIII (1761) RsrII (1778) MluI (1784) NotI (1794) PaeR7I - PspXI - XhoI (1802) PmeI (1811) FseI (1821) SacII (1825) XL39 (reverse primer) (1917 .. 1936) EcoO109I (2041) BbvCI - Bpu10I (2137) BlpI (2181) BsmBI (2201) AgeI (2399) PflMI (2402) XcmI (2494) SexAI * (2586) StuI (2818) AvrII (2819) BsaBI * (2888) KasI (3028) NarI (3029) SfoI (3030) PluTI (3032) PstI (3082) pCMV6-AN-FP635 6622 bp
DraIII  (6425)
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  (5872)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5753)
1 site
A G T A C T T C A T G A
PciI  (4380)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (4001)
1 site
G T A T A C C A T A T G
BsmI  (3949)
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  (3888)
2 sites
T T A T A A A A T A T T
PfoI  (3804)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BstBI  (3711)
1 site
T T C G A A A A G C T T
PflFI  (3147)
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  (3147)
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.
AanI  (6550)
2 sites
T T A T A A A A T A 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  (1250)
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.
Bsu36I  (1285)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
PshAI  (1659)
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  (1725)
1 site
T C C G G A A G G C C T
AsiSI  (1741)
1 site
G C G A T C G C C G C T A G C G
SgfI  (1741)
1 site
G C G A T C G C C G C T A G C G
MreI  (1743)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (1743)
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  (1747)
1 site
G G C G C G C C C C G C G C G G
HindIII  (1761)
1 site
A A G C T T T T C G A A
RsrII  (1778)
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  (1784)
1 site
A C G C G T T G C G C A