pFN28K HaloTag CMV-neo

Flexi® vector with a kanamycin/G418 resistance marker, for mammalian expression of a protein with a cleavable N-terminal HaloTag®.

Sequence Author: Promega

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ScaI (5495) MluI (5411) BspEI (5198) ApaLI (4748) PciI (4434) BstXI (4199) BstBI (4160) RsrII (3994) NaeI (3980) NgoMIV (3978) PluTI (3481) SfoI (3479) NarI (3478) KasI (3477) PacI (3265) AvrII (3241) SfiI (3194) SexAI * (3008) MfeI (2663) BglII (5530) BsrGI (96) SpeI (152) NdeI (387) SnaBI (493) T7 promoter NheI (1052) BmtI (1056) PflMI (1108) AleI (1166) BstEII (1184) BpmI (1239) BmgBI (1318) PasI (1348) BclI * (1548) AclI (1563) PshAI (1613) SgrAI (1700) PaeR7I - XhoI (1943) AsiSI - PvuI - SgfI (2001) DraI - PmeI (2368) EcoRI (2373) XbaI (2400) SbfI (2416) BlpI (2481) NotI (2511) AanI - AanI - PsiI (2634) HpaI (2654) pFN28K HaloTag® CMV-neo 5535 bp
ScaI  (5495)
1 site
A G T A C T T C A T G A
MluI  (5411)
1 site
A C G C G T T G C G C A
BspEI  (5198)
1 site
T C C G G A A G G C C T
ApaLI  (4748)
1 site
G T G C A C C A C G T G
PciI  (4434)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstXI  (4199)
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.
BstBI  (4160)
1 site
T T C G A A A A G C T T
RsrII  (3994)
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  (3980)
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  (3978)
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.
PluTI  (3481)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (3479)
1 site
G G C G C C C C G C G G
NarI  (3478)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (3477)
1 site
G G C G C C C C G C G G
PacI  (3265)
1 site
T T A A T T A A A A T T A A T T
AvrII  (3241)
1 site
C C T A G G G G A T C C
SfiI  (3194)
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.
SexAI  (3008)
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.
MfeI  (2663)
1 site
C A A T T G G T T A A C
BglII  (5530)
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.
SnaBI  (493)
1 site
T A C G T A A T G C A T
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.
BpmI  (1239)
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.
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.
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.
AclI  (1563)
1 site
A A C G T T T T G C A A
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.
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