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Plasmid Files

pFN18A HaloTag® T7

Flexi® vector with an ampicillin resistance marker, for bacterial or cell-free expression of a protein with a cleavable N-terminal HaloTag®.

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pFN18A HaloTag T7.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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PflMI (111) BglII (3929) MluI (3810) BspEI (3597) PciI (2833) AgeI (2765) BstBI (2738) AhdI (2663) FspI (2440) AleI (169) BstEII (187) BseRI (189) PflFI - Tth111I (319) BmgBI (321) PasI (351) StuI * (520) BclI * (551) BspDI * - ClaI * (557) PshAI (616) SgrAI (703) BbsI (902) BssHII (928) PaeR7I - XhoI (946) AsiSI - PvuI - SgfI (1004) BtgI - NcoI (1007) DraI - PmeI (1371) Acc65I (1388) KpnI (1392) BamHI (1397) XbaI (1403) PstI - SbfI (1419) BlpI (1484) EcoO109I (1511) SnaBI (1725) SspI (1858) XmnI (2063) pFN18A HaloTag® T7 4377 bp
PflMI  (111)
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  (3929)
1 site
A G A T C T T C T A G A
MluI  (3810)
1 site
A C G C G T T G C G C A
BspEI  (3597)
1 site
T C C G G A A G G C C T
PciI  (2833)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (2765)
1 site
A C C G G T T G G C C A
BstBI  (2738)
1 site
T T C G A A A A G C T T
AhdI  (2663)
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.
FspI  (2440)
1 site
T G C G C A A C G C G T
AleI  (169)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (187)
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  (189)
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  (319)
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  (319)
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  (321)
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  (351)
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  (520)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BclI  (551)
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  (557)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (557)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PshAI  (616)
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  (703)
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  (902)
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  (928)
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  (946)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (946)
1 site
C T C G A G G A G C T C
AsiSI  (1004)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1004)
1 site
C G A T C G G C T A G C
SgfI  (1004)
1 site
G C G A T C G C C G C T A G C G
BtgI  (1007)
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  (1007)
1 site
C C A T G G G G T A C C
DraI  (1371)
1 site
T T T A A A A A A T T T
PmeI  (1371)
1 site
G T T T A A A C C A A A T T T G
Acc65I  (1388)
1 site
G G T A C C C C A T G G
KpnI  (1392)
1 site
G G T A C C C C A T G G
BamHI  (1397)
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  (1403)
1 site
T C T A G A A G A T C T
PstI  (1419)
1 site
C T G C A G G A C G T C
SbfI  (1419)
1 site
C C T G C A G G G G A C G T C C
BlpI  (1484)
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  (1511)
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.
SnaBI  (1725)
1 site
T A C G T A A T G C A T
SspI  (1858)
1 site
A A T A T T T T A T A A
XmnI  (2063)
1 site
G A A N N N N T T C C T T N N N N A A G
HaloTag®
70 .. 960  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
HaloTag®
70 .. 960  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
AmpR
1876 .. 2736  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1876 .. 1944  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1876 .. 2736  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1945 .. 2736  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1876 .. 2736  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2894 .. 3482  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2894 .. 3482  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
1031 .. 1366  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
1031 .. 1366  =  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
3598 .. 3881  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3598 .. 3881  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
AmpR promoter
1771 .. 1875  =  105 bp
AmpR promoter
1771 .. 1875  =  105 bp
rrnB T1 terminator
4111 .. 4197  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
4111 .. 4197  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
1495 .. 1542  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1495 .. 1542  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
rrnB T2 terminator
4289 .. 4316  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
4289 .. 4316  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
TEV site
973 .. 993  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
TEV site
973 .. 993  =  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
ORF:  70 .. 1017  =  948 bp
ORF:  315 amino acids  =  35.7 kDa
ORF:  1876 .. 2736  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1031 .. 1366  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  4364 .. 268  =  282 bp
ORF:  93 amino acids  =  10.4 kDa
ORF:  3843 .. 4190  =  348 bp
ORF:  115 amino acids  =  12.4 kDa
ORF:  4348 .. 1032  =  1062 bp
ORF:  353 amino acids  =  39.9 kDa
ORF:  3952 .. 4278  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  633 .. 869  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  2340 .. 2606  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  848 .. 1087  =  240 bp
ORF:  79 amino acids  =  8.9 kDa
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