pFC27K HaloTag CMV-neo

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

Sequence Author: Promega

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ScaI (5487) MluI (5403) BspEI (5190) ApaLI (4740) PciI (4426) BstXI (4191) BstBI (4152) RsrII (3986) NaeI (3972) NgoMIV (3970) PluTI (3473) SfoI (3471) NarI (3470) KasI (3469) PacI (3257) AvrII (3233) SfiI (3186) SexAI * (3000) KpnI (2887) Acc65I (2883) BglII (5522) BsrGI (96) SpeI (152) CMV enhancer NdeI (387) SnaBI (493) T7 promoter NheI (1052) BmtI (1056) AsiSI - PvuI - SgfI (1060) AhdI (1436) Eco53kI - EcoICRI (1444) SacI (1446) PaeR7I - XhoI (1447) PflMI (1533) AleI (1591) BstEII (1609) BmgBI (1743) PasI (1773) BclI * (1973) AclI (1988) PshAI (2038) SgrAI (2125) EcoRI (2388) XbaI (2392) SalI (2398) SbfI (2408) BlpI (2473) NotI (2503) AanI - AanI - PsiI (2626) HpaI (2646) MfeI (2655) pFC27K HaloTag® CMV-neo 5527 bp
ScaI  (5487)
1 site
A G T A C T T C A T G A
MluI  (5403)
1 site
A C G C G T T G C G C A
BspEI  (5190)
1 site
T C C G G A A G G C C T
ApaLI  (4740)
1 site
G T G C A C C A C G T G
PciI  (4426)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstXI  (4191)
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  (4152)
1 site
T T C G A A A A G C T T
RsrII  (3986)
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  (3972)
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  (3970)
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  (3473)
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  (3471)
1 site
G G C G C C C C G C G G
NarI  (3470)
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  (3469)
1 site
G G C G C C C C G C G G
PacI  (3257)
1 site
T T A A T T A A A A T T A A T T
AvrII  (3233)
1 site
C C T A G G G G A T C C
SfiI  (3186)
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  (3000)
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.
KpnI  (2887)
1 site
G G T A C C C C A T G G
Acc65I  (2883)
1 site
G G T A C C C C A T G G
BglII  (5522)
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
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
AhdI  (1436)
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.
Eco53kI  (1444)
1 site
G A G C T C C T C G A G
EcoICRI  (1444)
1 site
G A G C T C C T C G A G
SacI  (1446)
1 site
G A G C T C C T C G A G
PaeR7I  (1447)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1447)
1 site
C T C G A G G A G C T C
PflMI  (1533)
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  (1591)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (1609)
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.
BmgBI  (1743)
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  (1773)
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  (1973)
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  (1988)
1 site
A A C G T T T T G C A A
PshAI  (2038)
1 site
G A C N N N N G T