Resources
Plasmid Files

pFC27A HaloTag® CMV-neo

Flexi® vector with ampicillin and G418 resistance markers, for mammalian expression of a protein with a cleavable C-terminal HaloTag®.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pFC27A HaloTag CMV-neo Sequence and MappFC27A HaloTag CMV-neo.dna
Map and Sequence File   
Sequence Author:  Promega
Download Free Trial Get SnapGene Viewer

 BglII (6519) MluI (6400) BspEI (6187) PciI (5423) AgeI (5355) XmnI (4653) poly(A) signal BstXI (4148) RsrII (3943) NaeI (3929) NgoMIV (3927) PluTI (3430) SfoI (3428) NarI (3427) KasI (3426) BsrGI (96) SpeI (152) CMV enhancer NdeI (387) T7 promoter NheI (1052) BmtI (1056) AsiSI - PvuI - SgfI (1060) Eco53kI - EcoICRI (1444) SacI (1446) PaeR7I - XhoI (1447) PflMI (1533) AleI (1591) BstEII (1609) BmgBI (1743) PasI (1773) BclI * (1973) PshAI (2038) SgrAI (2125) EcoRI (2388) XbaI (2392) SalI (2398) SbfI (2408) BlpI (2473) NotI (2503) AanI - PsiI (2626) HpaI (2646) MfeI (2655) Acc65I (2883) KpnI (2887) SexAI * (3000) SfiI (3186) AvrII (3233) pFC27A HaloTag® CMV-neo 6524 bp
BglII  (6519)
1 site
A G A T C T T C T A G A
MluI  (6400)
1 site
A C G C G T T G C G C A
BspEI  (6187)
1 site
T C C G G A A G G C C T
PciI  (5423)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (5355)
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.
XmnI  (4653)
1 site
G A A N N N N T T C C T T N N N N A A G
BstXI  (4148)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
RsrII  (3943)
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  (3929)
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  (3927)
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.
PluTI  (3430)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (3428)
1 site
G G C G C C C C G C G G
NarI  (3427)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
KasI  (3426)
1 site
G G C G C C C C G C G G
BsrGI  (96)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SpeI  (152)
1 site
A C T A G T T G A T C A
NdeI  (387)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
NheI  (1052)
1 site
G C T A G C C G A T C G
BmtI  (1056)
1 site
G C T A G C C G A T C G
AsiSI  (1060)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1060)
1 site
C G A T C G G C T A G C
SgfI  (1060)
1 site
G C G A T C G C C G C T A G C G
Eco53kI  (1444)
1 site
G A G C T C C T C G A G
EcoICRI  (1444)
1 site
G A G C T C C T C G A G
SacI  (1446)
1 site
G A G C T C C T C G A G
PaeR7I  (1447)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1447)
1 site
C T C G A G G A G C T C
PflMI  (1533)
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  (1591)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (1609)
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.
BmgBI  (1743)
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  (1773)
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.
BclI  (1973)
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.
PshAI  (2038)
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  (2125)
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.
EcoRI  (2388)
1 site
G A A T T C C T T A A G
XbaI  (2392)
1 site
T C T A G A A G A T C T
SalI  (2398)
1 site
G T C G A C C A G C T G
SbfI  (2408)
1 site
C C T G C A G G G G A C G T C C
BlpI  (2473)
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.
NotI  (2503)
1 site
G C G G C C G C C G C C G G C G
AanI  (2626)
1 site
T T A T A A A A T A T T
PsiI  (2626)
1 site
T T A T A A A A T A T T
HpaI  (2646)
1 site
G T T A A C C A A T T G
MfeI  (2655)
1 site
C A A T T G G T T A A C
Acc65I  (2883)
1 site
G G T A C C C C A T G G
KpnI  (2887)
1 site
G G T A C C C C A T G G
SexAI  (3000)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (3186)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AvrII  (3233)
1 site
C C T A G G G G A T C C
HaloTag®
1498 .. 2379  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
HaloTag®
1498 .. 2379  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
AmpR
4466 .. 5326  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4466 .. 4534  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4466 .. 5326  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4535 .. 5326  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4466 .. 5326  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
3299 .. 4093  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
3299 .. 4093  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
5484 .. 6072  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5484 .. 6072  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
2891 .. 3248  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2891 .. 3248  =  358 bp
SV40 enhancer and early promoter
barnase
1087 .. 1422  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus
amyloliquefaciens

The barnase gene is lethal in standard bacterial
transformation strains.
barnase
1087 .. 1422  =  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
6188 .. 6471  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
cer region
6188 .. 6471  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
2525 .. 2646  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2525 .. 2646  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4361 .. 4465  =  105 bp
AmpR promoter
4361 .. 4465  =  105 bp
poly(A) signal
4157 .. 4205  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4157 .. 4205  =  49 bp
synthetic polyadenylation signal
TEV site
1462 .. 1482  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV protease recognition site
TEV site
1462 .. 1482  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV protease recognition site
T7 promoter
1033 .. 1051  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1033 .. 1051  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
3099 .. 3234  =  136 bp
SV40 origin of replication
SV40 ori
3099 .. 3234  =  136 bp
SV40 origin of replication
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file 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.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter