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pH6HTN His6HaloTag® T7

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

To see this sequence with restriction sites, features, and translations, please download
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pH6HTN His6HaloTag T7.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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PflMI (132) BglII (3566) MluI (3447) BspEI (3234) PciI (2470) AgeI (2402) BstBI (2375) AhdI (2300) NmeAIII (2153) SphI (156) AleI (190) BstEII (208) BseRI (210) PflFI - Tth111I (340) BmgBI (342) PasI (372) StuI * (541) BclI * (572) BspDI * - ClaI * (578) PshAI (637) SgrAI (724) TspMI - XmaI (894) SmaI (896) BbsI (923) BssHII (949) SalI (960) AccI (961) PaeR7I - XhoI (967) AsiSI - PvuI (1025) Eco53kI (1042) SacI (1044) BtgI (1048) SacII (1051) EcoRV (1055) XbaI (1057) PspOMI (1066) ApaI (1070) PstI - SbfI (1081) NotI (1090) BlpI (1121) StyI (1143) EcoO109I (1148) BfuAI - BspMI (1273) SnaBI (1362) SspI (1495) XmnI (1700) FspI (2077) pH6HTN His6HaloTag® T7 4014 bp
PflMI  (132)
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.
BglII  (3566)
1 site
A G A T C T T C T A G A
MluI  (3447)
1 site
A C G C G T T G C G C A
BspEI  (3234)
1 site
T C C G G A A G G C C T
PciI  (2470)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (2402)
1 site
A C C G G T T G G C C A
BstBI  (2375)
1 site
T T C G A A A A G C T T
AhdI  (2300)
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  (2153)
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).
SphI  (156)
1 site
G C A T G C C G T A C G
AleI  (190)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (208)
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  (210)
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  (340)
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  (340)
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  (342)
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  (372)
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  (541)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BclI  (572)
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  (578)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (578)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PshAI  (637)
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  (724)
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  (894)
1 site
C C C G G G G G G C C C
XmaI  (894)
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  (896)
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  (923)
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  (949)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
SalI  (960)
1 site
G T C G A C C A G C T G
AccI  (961)
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  (967)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (967)
1 site
C T C G A G G A G C T C
AsiSI  (1025)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1025)
1 site
C G A T C G G C T A G C
Eco53kI  (1042)
1 site
G A G C T C C T C G A G
SacI  (1044)
1 site
G A G C T C C T C G A G
BtgI  (1048)
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  (1051)
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  (1055)
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  (1057)
1 site
T C T A G A A G A T C T
PspOMI  (1066)
1 site
G G G C C C C C C G G G
ApaI  (1070)
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  (1081)
1 site
C T G C A G G A C G T C
SbfI  (1081)
1 site
C C T G C A G G G G A C G T C C
NotI  (1090)
1 site
G C G G C C G C C G C C G G C G
BlpI  (1121)
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.
StyI  (1143)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
EcoO109I  (1148)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
BfuAI  (1273)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1273)
1 site
A C C T G C ( N ) 4 T