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Plasmid Files

pFC16A HaloTag® CMVd2

Flexi® vector with an ampicillin resistance marker, for mammalian expression from a moderately compromised CMV promoter of a protein with a cleavable C-terminal HaloTag®.

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pFC16A HaloTag CMVd2 Sequence and MappFC16A HaloTag CMVd2.dna
Map and Sequence File   
Sequence Author:  Promega
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 BsaBI * (4128) MluI (4010) BspEI (3797) PciI (3033) AgeI (2965) BstBI (2938) AseI (2688) FspI (2640) XmnI (2263) SspI (2058) BglII (4129) HindIII (79) AsiSI - PvuI - SgfI (138) BtgI - NcoI (141) Eco53kI - EcoICRI (522) SacI (524) PaeR7I - XhoI (525) PflMI (611) AleI (669) BstEII (687) BseRI (689) PflFI - Tth111I (819) BmgBI (821) PasI (851) StuI * (1020) BclI * (1051) BspDI * - ClaI * (1057) PshAI (1116) SgrAI (1203) BbsI (1402) BssHII (1428) XbaI (1470) SalI (1476) PstI - SbfI (1486) BfuAI - BspMI (1489) BlpI (1551) NotI (1581) AanI - PsiI (1704) HpaI (1724) MfeI (1733) SnaBI (1925) pFC16A HaloTag® CMVd2 4134 bp
BsaBI  (4128)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
MluI  (4010)
1 site
A C G C G T T G C G C A
BspEI  (3797)
1 site
T C C G G A A G G C C T
PciI  (3033)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (2965)
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.
BstBI  (2938)
1 site
T T C G A A A A G C T T
AseI  (2688)
1 site
A T T A A T T A A T T A
FspI  (2640)
1 site
T G C G C A A C G C G T
XmnI  (2263)
1 site
G A A N N N N T T C C T T N N N N A A G
SspI  (2058)
1 site
A A T A T T T T A T A A
BglII  (4129)
1 site
A G A T C T T C T A G A
HindIII  (79)
1 site
A A G C T T T T C G A A
AsiSI  (138)
1 site
G C G A T C G C C G C T A G C G
PvuI  (138)
1 site
C G A T C G G C T A G C
SgfI  (138)
1 site
G C G A T C G C C G C T A G C G
BtgI  (141)
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  (141)
1 site
C C A T G G G G T A C C
Eco53kI  (522)
1 site
G A G C T C C T C G A G
EcoICRI  (522)
1 site
G A G C T C C T C G A G
SacI  (524)
1 site
G A G C T C C T C G A G
PaeR7I  (525)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (525)
1 site
C T C G A G G A G C T C
PflMI  (611)
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  (669)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (687)
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  (689)
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  (819)
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  (819)
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  (821)
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  (851)
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  (1020)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BclI  (1051)
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  (1057)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1057)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PshAI  (1116)
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  (1203)
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  (1402)
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  (1428)
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.
XbaI  (1470)
1 site
T C T A G A A G A T C T
SalI  (1476)
1 site
G T C G A C C A G C T G
PstI  (1486)
1 site
C T G C A G G A C G T C
SbfI  (1486)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (1489)
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  (1489)
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.
BlpI  (1551)
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  (1581)
1 site
G C G G C C G C C G C C G G C G
AanI  (1704)
1 site
T T A T A A A A T A T T
PsiI  (1704)
1 site
T T A T A A A A T A T T
HpaI  (1724)
1 site
G T T A A C C A A T T G
MfeI  (1733)
1 site
C A A T T G G T T A A C
SnaBI  (1925)
1 site
T A C G T A A T G C A T
HaloTag®
576 .. 1457  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
HaloTag®
576 .. 1457  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
AmpR
2076 .. 2936  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2076 .. 2144  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2076 .. 2936  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2145 .. 2936  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2076 .. 2936  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3094 .. 3682  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3094 .. 3682  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
barnase
165 .. 500  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus
amyloliquefaciens

The barnase gene is lethal in standard bacterial
transformation strains.
barnase
165 .. 500  =  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
3798 .. 4081  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
cer region
3798 .. 4081  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
SV40 poly(A) signal
1603 .. 1724  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1603 .. 1724  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1971 .. 2075  =  105 bp
AmpR promoter
1971 .. 2075  =  105 bp
CMVd2 promoter
1 .. 52  =  52 bp
truncated human cytomegalovirus (CMV) immediate
early promoter that yields moderately reduced
expression relative to the full-length promoter
CMVd2 promoter
1 .. 52  =  52 bp
truncated human cytomegalovirus (CMV) immediate
early promoter that yields moderately reduced
expression relative to the full-length promoter
TEV site
540 .. 560  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV protease recognition site
TEV site
540 .. 560  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV protease recognition site
T7 promoter
84 .. 102  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
84 .. 102  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
108 .. 126  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
108 .. 126  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
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