pHTN HaloTag CMV-neo

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

Sequence Author: Promega

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BglII (6138) MluI (6019) BspEI (5806) PciI (5042) AgeI (4974) AhdI (4872) XmnI (4272) BamHI (3840) poly(A) signal BstXI (3767) RsrII (3562) NaeI (3548) NgoMIV (3546) PluTI (3049) BsrGI (96) SpeI (152) CMV enhancer NdeI (387) T7 promoter NheI (1052) BmtI (1056) PflMI (1108) AleI (1166) BstEII (1184) BmgBI (1318) PasI (1348) BclI * (1548) PshAI (1613) SgrAI (1700) TspMI - XmaI (1870) SmaI (1872) SalI (1936) AccI (1937) PaeR7I - XhoI (1943) TEV site AsiSI - PvuI (2001) EcoRI (2009) Eco53kI (2018) SacI (2020) SacII (2027) EcoRV (2031) XbaI (2033) PspOMI (2042) ApaI (2046) SbfI (2057) NotI (2066) BlpI (2097) PsiI (2245) HpaI (2265) MfeI (2274) Acc65I (2502) KpnI (2506) CsiI - SexAI * (2619) SV40 promoter SfiI (2805) AvrII (2852) KasI (3045) NarI (3046) SfoI (3047) pHTN HaloTag® CMV-neo 6143 bp
BglII  (6138)
1 site
A G A T C T T C T A G A
MluI  (6019)
1 site
A C G C G T T G C G C A
BspEI  (5806)
1 site
T C C G G A A G G C C T
PciI  (5042)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (4974)
1 site
A C C G G T T G G C C A
AhdI  (4872)
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  (4272)
1 site
G A A N N N N T T C C T T N N N N A A G
BamHI  (3840)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BstXI  (3767)
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  (3562)
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  (3548)
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  (3546)
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  (3049)
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.
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
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.
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.
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.
TspMI  (1870)
1 site
C C C G G G G G G C C C
XmaI  (1870)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (1872)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SalI  (1936)
1 site
G T C G A C C A G C T G
AccI  (1937)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
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
EcoRI  (2009)
1 site
G A A T T C C T T A A G
Eco53kI  (2018)
1 site
G A G C T C C T C G A G
SacI  (2020)
1 site
G A G C T C C T C G A G
SacII  (2027)
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  (2031)
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  (2033)
1 site
T C T A G A A G A T C T
PspOMI  (2042)
1 site
G G G C C C C C C G G G
ApaI  (2046)
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  (2057)
1 site
C C T G C A G G G G A C G T C C
NotI  (2066)
1 site
G C G G C C G C C G C C G G C G
BlpI  (2097)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
PsiI  (2245)
1 site
T T A T A A A A T A T T
HpaI  (2265)
1 site
G T T A A C C A A T T G
MfeI  (2274)
1 site
C A A T T G G T T A A C
Acc65I  (2502)
1 site
G G T A C C C C A T G G
KpnI  (2506)
1 site
G G T A C C C C A T G G
CsiI  (2619)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (2619)
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.
SfiI  (2805)
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.
AvrII  (2852)
1 site
C C T A G G G G A T C C
KasI  (3045)
1 site
G G C G C C C C G C G G
NarI  (3046)
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.
SfoI  (3047)
1 site
G G C G C C C C G C G G
HaloTag®