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pH6HTC His6HaloTag® T7

Vector with a traditional MCS, for bacterial or cell-free expression of a protein with a cleavable C-terminal HaloTag®-6xHis dual tag.

To see this sequence with restriction sites, features, and translations, please download
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pH6HTC His6HaloTag T7.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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BglII (3536) MluI (3417) BspEI (3204) PciI (2440) AgeI (2372) BstBI (2345) AhdI (2270) NmeAIII (2123) FspI (2047) AsiSI - PvuI (65) EcoRI (73) BtgI (82) SacII (85) EcoRV (89) XbaI (91) PspOMI (100) ApaI (104) PstI - SbfI (115) Eco53kI (121) SacI (123) PaeR7I - XhoI (124) BamHI (169) PflMI (210) SphI (234) AleI (268) BstEII (286) BseRI (288) PflFI - Tth111I (418) BmgBI (420) PasI (450) StuI * (619) BclI * (650) BspDI * - ClaI * (656) PshAI (715) SgrAI (802) TspMI - XmaI (972) SmaI (974) BbsI (1001) BssHII (1027) AccI (1039) BlpI (1091) StyI (1113) EcoO109I (1118) BfuAI - BspMI (1243) SnaBI (1332) SspI (1465) XmnI (1670) pH6HTC His6HaloTag® T7 3984 bp
BglII  (3536)
1 site
A G A T C T T C T A G A
MluI  (3417)
1 site
A C G C G T T G C G C A
BspEI  (3204)
1 site
T C C G G A A G G C C T
PciI  (2440)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (2372)
1 site
A C C G G T T G G C C A
BstBI  (2345)
1 site
T T C G A A A A G C T T
AhdI  (2270)
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.
NmeAIII  (2123)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
FspI  (2047)
1 site
T G C G C A A C G C G T
AsiSI  (65)
1 site
G C G A T C G C C G C T A G C G
PvuI  (65)
1 site
C G A T C G G C T A G C
EcoRI  (73)
1 site
G A A T T C C T T A A G
BtgI  (82)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SacII  (85)
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  (89)
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  (91)
1 site
T C T A G A A G A T C T
PspOMI  (100)
1 site
G G G C C C C C C G G G
ApaI  (104)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PstI  (115)
1 site
C T G C A G G A C G T C
SbfI  (115)
1 site
C C T G C A G G G G A C G T C C
Eco53kI  (121)
1 site
G A G C T C C T C G A G
SacI  (123)
1 site
G A G C T C C T C G A G
PaeR7I  (124)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (124)
1 site
C T C G A G G A G C T C
BamHI  (169)
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.
PflMI  (210)
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.
SphI  (234)
1 site
G C A T G C C G T A C G
AleI  (268)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (286)
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  (288)
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  (418)
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  (418)
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  (420)
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  (450)
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  (619)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BclI  (650)
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  (656)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (656)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PshAI  (715)
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  (802)
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  (972)
1 site
C C C G G G G G G C C C
XmaI  (972)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (974)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BbsI  (1001)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BssHII  (1027)
1 site
G C G C G C C G C G C