pCMV6-AC-mRFP

PrecisionShuttle™ mammalian expression vector for fusing an ORF to a C-terminal TagRFP tag.

Sequence Author: OriGene

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AanI (6556) XmnI (5878) ScaI (5759) PciI (4386) BstZ17I (4007) BsmI (3955) AanI (3894) BstBI (3717) 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 (1030) HindIII (1044) RsrII (1061) MluI (1067) BsiWI (1070) NotI (1076) PaeR7I - XhoI (1082) PasI (1318) Bsu36I (1353) PshAI (1727) BspEI (1799) PmeI (1817) FseI (1827) SacII (1831) AleI (1917) XL39 (reverse primer) (1923 .. 1942) EcoO109I (2047) BsgI (2101) BbvCI - Bpu10I (2143) BlpI (2187) BsmBI (2207) BstXI (2294) AgeI (2405) XcmI (2500) SexAI * (2592) SfiI (2778) StuI (2824) AvrII (2825) BclI * (2876) BsaBI * (2894) KasI (3034) NarI (3035) SfoI (3036) PluTI (3038) PstI (3088) PflFI - Tth111I (3153) pCMV6-AC-mRFP 6628 bp
AanI  (6556)
2 sites
T T A T A A A A T A T T
XmnI  (5878)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5759)
1 site
A G T A C T T C A T G A
PciI  (4386)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (4007)
1 site
G T A T A C C A T A T G
BsmI  (3955)
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  (3894)
2 sites
T T A T A A A A T A T T
BstBI  (3717)
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
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
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
BsiWI  (1070)
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.
NotI  (1076)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1082)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1082)
1 site
C T C G A G G A G C T C
PasI  (1318)
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  (1353)
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  (1727)
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  (1799)
1 site
T C C G G A A G G C C T
PmeI  (1817)
1 site
G T T T A A A C C A A A T T T G
FseI  (1827)
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  (1831)
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  (1917)
1 site
C A C N N N N G T G G T G N N N N C A C
EcoO109I  (2047)
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  (2101)
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  (2143)
1 site
C C T C A G C G G A G T C G
Bpu10I  (2143)
1 site
C C T N A G C G