pFN29K His6HaloTag T7

Flexi® vector with a kanamycin resistance marker, 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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AseI (20) ScaI (3910) MluI (3826) BspEI (3613) ApaLI (3163) PciI (2849) AgeI (2781) BstBI (2754) RsrII (2588) NaeI (2574) NgoMIV (2572) T7 promoter ATG 6xHis PflMI (132) AleI (190) BstEII (208) BseRI (210) BpmI (263) BmgBI (342) PasI (372) StuI * (541) BspDI * - ClaI * (578) PshAI (637) BglI (696) SgrAI (724) BbsI (923) BssHII (949) PaeR7I - XhoI (967) AsiSI - PvuI - SgfI (1025) BtgI - NcoI (1028) DraI - PmeI (1392) Acc65I (1409) KpnI (1413) BamHI (1418) XbaI (1424) SbfI (1440) BlpI (1505) EcoO109I (1532) BtsαI (1636) BstAPI (1770) XcmI (1888) FspI (2174) pFN29K His6HaloTag® T7 4393 bp
AseI  (20)
1 site
A T T A A T T A A T T A
ScaI  (3910)
1 site
A G T A C T T C A T G A
MluI  (3826)
1 site
A C G C G T T G C G C A
BspEI  (3613)
1 site
T C C G G A A G G C C T
ApaLI  (3163)
1 site
G T G C A C C A C G T G
PciI  (2849)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (2781)
1 site
A C C G G T T G G C C A
BstBI  (2754)
1 site
T T C G A A A A G C T T
RsrII  (2588)
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  (2574)
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  (2572)
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.
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.
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.
BpmI  (263)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
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.
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.
BglI  (696)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
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.
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.
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
SgfI  (1025)
1 site
G C G A T C G C C G C T A G C G
BtgI  (1028)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1028)
1 site
C C A T G G G G T A C C
DraI  (1392)
1 site
T T T A A A A A A T T T
PmeI  (1392)
1 site
G T T T A A A C C A A A T T T G
Acc65I  (1409)
1 site
G G T A C C C C A T G G
KpnI  (1413)
1 site
G G T A C C C C A T G G
BamHI  (1418)
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.
XbaI  (1424)
1 site
T C T A G A A G A T C T
SbfI  (1440)
1 site
C C T G C A G G G G A C G T C C
BlpI  (1505)
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.
EcoO109I  (1532)
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.
BtsαI  (1636)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
BstAPI  (1770)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
XcmI  (1888)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
FspI  (2174)
1 site
T G C G C A A C G C G T
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
NeoR/KanR
1944 .. 2738  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
1944 .. 2738  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
2910 .. 3498  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2910 .. 3498  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
1052 .. 1387  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
1052 .. 1387  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
cer region
3614 .. 3897  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3614 .. 3897  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
rrnB T1 terminator
4127 .. 4213  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
4127 .. 4213  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
1516 .. 1563  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1516 .. 1563  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
rrnB T2 terminator
4305 .. 4332  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
4305 .. 4332  =  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 .. 1038  =  969 bp
ORF:  322 amino acids  =  36.6 kDa
ORF:  2116 .. 2502  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  3859 .. 4206  =  348 bp
ORF:  115 amino acids  =  12.4 kDa
ORF:  1052 .. 1387  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  1944 .. 2738  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  869 .. 1108  =  240 bp
ORF:  79 amino acids  =  8.9 kDa
ORF:  3968 .. 4294  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  4364 .. 1053  =  1083 bp
ORF:  360 amino acids  =  40.8 kDa
ORF:  654 .. 890  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  2253 .. 2507  =  255 bp
ORF:  84 amino acids  =  9.8 kDa
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Download pFN29K His6HaloTag T7.dna file

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