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Plasmid Files

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.

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pH6HTN His6HaloTag T7 Sequence and MappH6HTN His6HaloTag T7.dna
Map and Sequence File   
Sequence Author:  Promega
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 T7 promoter BglII (3566) MluI (3447) BspEI (3234) PciI (2470) AgeI (2402) BstBI (2375) AhdI (2300) NmeAIII (2153) ATG 6xHis PflMI (132) 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
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

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
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).
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.
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 G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
SnaBI  (1362)
1 site
T A C G T A A T G C A T
SspI  (1495)
1 site
A A T A T T T T A T A A
XmnI  (1700)
1 site
G A A N N N N T T C C T T N N N N A A G
FspI  (2077)
1 site
T G C G C A A C G C G T
HaloTag®
94 .. 981  =  888 bp
296 amino acids  =  33.5 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
HaloTag®
94 .. 981  =  888 bp
296 amino acids  =  33.5 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
AmpR
1513 .. 2373  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1513 .. 1581  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1513 .. 2373  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1582 .. 2373  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1513 .. 2373  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2531 .. 3119  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2531 .. 3119  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
cer region
3235 .. 3518  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
cer region
3235 .. 3518  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
AmpR promoter
1408 .. 1512  =  105 bp
AmpR promoter
1408 .. 1512  =  105 bp
rrnB T1 terminator
3748 .. 3834  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
3748 .. 3834  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
MCS
1021 .. 1096  =  76 bp
multiple cloning site
MCS
1021 .. 1096  =  76 bp
multiple cloning site
T7 terminator
1132 .. 1179  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
1132 .. 1179  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
rrnB T2 terminator
3926 .. 3953  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
3926 .. 3953  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
TEV site
994 .. 1014  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV protease recognition site
TEV site
994 .. 1014  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV protease recognition site
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
76 .. 93  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
76 .. 93  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
70 .. 72  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
70 .. 72  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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