pFC20A HaloTag T7 SP6

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

Sequence Author: Promega

|Download SnapGene Viewer
Explore Over 2.7k Plasmids: Basic Cloning Vectors | More Plasmid Sets
No matches
T7 promoter BglII (3974) MluI (3855) BspEI (3642) PciI (2878) AgeI (2810) BstBI (2783) FspI (2485) SP6 promoter AsiSI - PvuI - SgfI (75) BtgI - NcoI (78) Eco53kI - EcoICRI (459) SacI (461) PaeR7I - XhoI (462) BamHI (507) PflMI (548) AleI (606) BstEII (624) BseRI (626) PflFI - Tth111I (756) BmgBI (758) PasI (788) StuI * (957) BclI * (988) BspDI * - ClaI * (994) PshAI (1053) SgrAI (1140) BbsI (1339) BssHII (1365) EcoRI (1403) XbaI (1407) SalI (1413) PstI - SbfI (1423) BlpI (1488) EcoO109I (1556) SnaBI (1770) SspI (1903) XmnI (2108) pFC20A HaloTag® T7 SP6 4422 bp
BglII  (3974)
1 site
A G A T C T T C T A G A
MluI  (3855)
1 site
A C G C G T T G C G C A
BspEI  (3642)
1 site
T C C G G A A G G C C T
PciI  (2878)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (2810)
1 site
A C C G G T T G G C C A
BstBI  (2783)
1 site
T T C G A A A A G C T T
FspI  (2485)
1 site
T G C G C A A C G C G T
AsiSI  (75)
1 site
G C G A T C G C C G C T A G C G
PvuI  (75)
1 site
C G A T C G G C T A G C
SgfI  (75)
1 site
G C G A T C G C C G C T A G C G
BtgI  (78)
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  (78)
1 site
C C A T G G G G T A C C
Eco53kI  (459)
1 site
G A G C T C C T C G A G
EcoICRI  (459)
1 site
G A G C T C C T C G A G
SacI  (461)
1 site
G A G C T C C T C G A G
PaeR7I  (462)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (462)
1 site
C T C G A G G A G C T C
BamHI  (507)
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.
PflMI  (548)
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  (606)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (624)
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  (626)
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  (756)
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  (756)
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  (758)
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  (788)
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  (957)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BclI  (988)
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  (994)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (994)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PshAI  (1053)
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  (1140)
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  (1339)
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  (1365)
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.
EcoRI  (1403)
1 site
G A A T T C C T T A A G
XbaI  (1407)
1 site
T C T A G A A G A T C T
SalI  (1413)
1 site
G T C G A C C A G C T G
PstI  (1423)
1 site
C T G C A G G A C G T C
SbfI  (1423)
1 site
C C T G C A G G G G A C G T C C
BlpI  (1488)
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  (1556)
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  (1770)
1 site
T A C G T A A T G C A T
SspI  (1903)
1 site
A A T A T T T T A T A A
XmnI  (2108)
1 site
G A A N N N N T T C C T T N N N N A A G
TEV site
477 .. 497  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
TEV site
477 .. 497  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
HaloTag®
513 .. 1394  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
HaloTag®
513 .. 1394  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
AmpR
1921 .. 2781  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1921 .. 1989  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1921 .. 2781  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1990 .. 2781  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1921 .. 2781  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2939 .. 3527  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2939 .. 3527  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
102 .. 437  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
102 .. 437  =  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
3643 .. 3926  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3643 .. 3926  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
AmpR promoter
1816 .. 1920  =  105 bp
AmpR promoter
1816 .. 1920  =  105 bp
rrnB T1 terminator
4156 .. 4242  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
4156 .. 4242  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
1540 .. 1587  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1540 .. 1587  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
rrnB T2 terminator
4334 .. 4361  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
4334 .. 4361  =  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
SP6 promoter
45 .. 63  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
45 .. 63  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
ORF:  1921 .. 2781  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  102 .. 437  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  444 .. 1400  =  957 bp
ORF:  318 amino acids  =  36.0 kDa
ORF:  3888 .. 4235  =  348 bp
ORF:  115 amino acids  =  12.4 kDa
ORF:  3997 .. 4323  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  441 .. 1070  =  630 bp
ORF:  209 amino acids  =  23.0 kDa
ORF:  2385 .. 2651  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1070 .. 1306  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
Click here to try SnapGene

Download pFC20A HaloTag T7 SP6.dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

Individual Sequences & Maps

The maps, notes, and annotations in the zip file on this page are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as ’’www.snapgene.com/resources’’. Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Discover the most user-friendly molecular biology experience.