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.

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) FspI (2077) 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) 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
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).
FspI  (2077)
1 site
T G C G C A A C G C G T
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

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
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
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
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
TEV site
994 .. 1014  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
TEV site
994 .. 1014  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
AmpR
1513 .. 2373  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   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
2 segments
   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
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  70 .. 1131  =  1062 bp
ORF:  353 amino acids  =  40.0 kDa
ORF:  1513 .. 2373  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3480 .. 3827  =  348 bp
ORF:  115 amino acids  =  12.4 kDa
ORF:  3985 .. 654  =  684 bp
ORF:  227 amino acids  =  25.3 kDa
ORF:  3589 .. 3915  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  654 .. 890  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  1977 .. 2243  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Download pH6HTN His6HaloTag T7.dna file

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