pHTC HaloTag CMV-neo

Mammalian expression vector with a traditional MCS and ampicillin and G418 resistance markers, encoding a cleavable C-terminal HaloTag®.

Sequence Author: Promega

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BglII (6180) MluI (6061) BspEI (5848) PciI (5084) AgeI (5016) AhdI (4914) XmnI (4314) BstXI (3809) RsrII (3604) NaeI (3590) NgoMIV (3588) PluTI (3091) SfoI (3089) BsrGI (96) SpeI (152) NdeI (387) T7 promoter NheI (1052) BmtI (1056) AsiSI - PvuI (1065) EcoRI (1073) SacII (1085) EcoRV (1089) MCS XbaI (1091) PspOMI (1100) ApaI (1104) SbfI (1115) Eco53kI (1121) SacI (1123) PaeR7I - XhoI (1124) TEV site PflMI (1210) AleI (1268) BstEII (1286) BmgBI (1420) PasI (1450) BclI * (1650) PshAI (1715) SgrAI (1802) TspMI - XmaI (1972) SmaI (1974) AccI (2039) BlpI (2134) NotI (2164) PsiI (2287) HpaI (2307) MfeI (2316) Acc65I (2544) KpnI (2548) CsiI - SexAI * (2661) SfiI (2847) AvrII (2894) KasI (3087) NarI (3088) pHTC HaloTag® CMV-neo 6185 bp
BglII  (6180)
1 site
A G A T C T T C T A G A
MluI  (6061)
1 site
A C G C G T T G C G C A
BspEI  (5848)
1 site
T C C G G A A G G C C T
PciI  (5084)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (5016)
1 site
A C C G G T T G G C C A
AhdI  (4914)
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.
XmnI  (4314)
1 site
G A A N N N N T T C C T T N N N N A A G
BstXI  (3809)
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  (3604)
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  (3590)
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  (3588)
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  (3091)
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  (3089)
1 site
G G C G C C C C G C G G
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.
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  (1065)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1065)
1 site
C G A T C G G C T A G C
EcoRI  (1073)
1 site
G A A T T C C T T A A G
SacII  (1085)
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.
EcoRV  (1089)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
XbaI  (1091)
1 site
T C T A G A A G A T C T
PspOMI  (1100)
1 site
G G G C C C C C C G G G
ApaI  (1104)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SbfI  (1115)
1 site
C C T G C A G G G G A C G T C C
Eco53kI  (1121)
1 site
G A G C T C C T C G A G
SacI  (1123)
1 site
G A G C T C C T C G A G
PaeR7I  (1124)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1124)
1 site
C T C G A G G A G C T C
PflMI  (1210)
1 site
C C A N N N N N T G G G G T N N