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pFN21A HaloTag® CMV

Flexi® vector with an ampicillin resistance marker, for mammalian expression of a protein with a cleavable N-terminal HaloTag®.

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pFN21A HaloTag CMV.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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BsaBI * (5058) MluI (4940) BspEI (4727) PciI (3963) AgeI (3895) BstBI (3868) AhdI (3793) FspI (3570) XmnI (3193) MfeI (2663) HpaI (2654) AanI - PsiI (2634) BglII (5059) BsrGI (96) SpeI (152) NdeI (387) HindIII (748) BstAPI (794) NheI (1052) BmtI (1056) PflMI (1108) AleI (1166) BstEII (1184) BseRI (1186) PflFI - Tth111I (1316) BmgBI (1318) PasI (1348) StuI * (1517) BclI * (1548) BspDI * - ClaI * (1554) PshAI (1613) SgrAI (1700) BssHII (1925) PaeR7I - XhoI (1943) AsiSI - PvuI - SgfI (2001) DraI - PmeI (2368) Acc65I (2385) KpnI (2389) XbaI (2400) SbfI (2416) BlpI (2481) NotI (2511) pFN21A HaloTag® CMV 5064 bp
BsaBI  (5058)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
MluI  (4940)
1 site
A C G C G T T G C G C A
BspEI  (4727)
1 site
T C C G G A A G G C C T
PciI  (3963)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (3895)
1 site
A C C G G T T G G C C A
BstBI  (3868)
1 site
T T C G A A A A G C T T
AhdI  (3793)
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.
FspI  (3570)
1 site
T G C G C A A C G C G T
XmnI  (3193)
1 site
G A A N N N N T T C C T T N N N N A A G
MfeI  (2663)
1 site
C A A T T G G T T A A C
HpaI  (2654)
1 site
G T T A A C C A A T T G
AanI  (2634)
1 site
T T A T A A A A T A T T
PsiI  (2634)
1 site
T T A T A A A A T A T T
BglII  (5059)
1 site
A G A T C T T C T A G A
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.
HindIII  (748)
1 site
A A G C T T T T C G A A
BstAPI  (794)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
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
PflMI  (1108)
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.
AleI  (1166)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (1184)
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.
BseRI  (1186)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
PflFI  (1316)
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  (1316)
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.
BmgBI  (1318)
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.
PasI  (1348)
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.
StuI  (1517)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BclI  (1548)
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.
BspDI  (1554)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1554)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PshAI  (1613)
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.
SgrAI  (1700)
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.
BssHII  (1925)
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.
PaeR7I  (1943)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1943)
1 site
C T C G A G G A G C T C
AsiSI  (2001)
1 site
G C G A T C G C C G C T A G C G
PvuI  (2001)
1 site
C G A T C G G C T A G C
SgfI  (2001)
1 site
G C G A T C G C C G C T A G C G
DraI  (2368)
1 site
T T T