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pF5A CMV-neo

Flexi® vector with a ampicillin and G418 resistance markers, for protein expression in mammalian cells.

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pF5A CMV-neo.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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BglII (5586) MluI (5467) BspEI (5254) PciI (4490) AgeI (4422) AhdI (4320) BpmI (4251) XmnI (3720) BspDI - ClaI (3275) BstXI (3215) RsrII (3010) NaeI (2996) NgoMIV (2994) BssHII (2891) BsrGI (96) SpeI (152) NdeI (387) BbsI (928) NheI (1052) BmtI (1056) AsiSI - PvuI - SgfI (1060) DraI - PmeI (1427) EcoRI (1432) XbaI (1459) SalI (1465) AccI (1466) SbfI (1475) BlpI (1540) NotI (1570) PsiI (1693) HpaI (1713) MfeI (1722) SexAI * (2067) SfiI (2253) BseRI (2296) StuI (2299) AvrII (2300) KasI (2493) NarI (2494) SfoI (2495) PluTI (2497) PflFI - Tth111I (2612) pF5A CMV-neo 5591 bp
BglII  (5586)
1 site
A G A T C T T C T A G A
MluI  (5467)
1 site
A C G C G T T G C G C A
BspEI  (5254)
1 site
T C C G G A A G G C C T
PciI  (4490)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (4422)
1 site
A C C G G T T G G C C A
AhdI  (4320)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BpmI  (4251)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
XmnI  (3720)
1 site
G A A N N N N T T C C T T N N N N A A G
BspDI  (3275)
1 site
A T C G A T T A G C T A
ClaI  (3275)
1 site
A T C G A T T A G C T A
BstXI  (3215)
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.
RsrII  (3010)
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.
NaeI  (2996)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (2994)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
BssHII  (2891)
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.
BsrGI  (96)
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.
SpeI  (152)
1 site
A C T A G T T G A T C A
NdeI  (387)
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.
BbsI  (928)
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.
NheI  (1052)
1 site
G C T A G C C G A T C G
BmtI  (1056)
1 site
G C T A G C C G A T C G
AsiSI  (1060)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1060)
1 site
C G A T C G G C T A G C
SgfI  (1060)
1 site
G C G A T C G C C G C T A G C G
DraI  (1427)
1 site
T T T A A A A A A T T T
PmeI  (1427)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (1432)
1 site
G A A T T C C T T A A G
XbaI  (1459)
1 site
T C T A G A A G A T C T
SalI  (1465)
1 site
G T C G A C C A G C T G
AccI  (1466)
1 site