pFC15A HaloTag CMVd1

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

Sequence Author: Promega

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BsaBI * (4176) MluI (4058) BspEI (3845) PciI (3081) AgeI (3013) BstBI (2986) AseI (2736) FspI (2688) XmnI (2311) SspI (2106) BglII (4177) HindIII (127) AsiSI - PvuI - SgfI (186) BtgI - NcoI (189) Eco53kI - EcoICRI (570) SacI (572) PaeR7I - XhoI (573) PflMI (659) AleI (717) BstEII (735) BseRI (737) PflFI - Tth111I (867) BmgBI (869) PasI (899) StuI * (1068) BclI * (1099) BspDI * - ClaI * (1105) PshAI (1164) SgrAI (1251) BbsI (1450) BssHII (1476) XbaI (1518) SalI (1524) PstI - SbfI (1534) BfuAI - BspMI (1537) BlpI (1599) NotI (1629) AanI - PsiI (1752) HpaI (1772) MfeI (1781) SnaBI (1973) pFC15A HaloTag® CMVd1 4182 bp
BsaBI  (4176)
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  (4058)
1 site
A C G C G T T G C G C A
BspEI  (3845)
1 site
T C C G G A A G G C C T
PciI  (3081)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (3013)
1 site
A C C G G T T G G C C A
BstBI  (2986)
1 site
T T C G A A A A G C T T
AseI  (2736)
1 site
A T T A A T T A A T T A
FspI  (2688)
1 site
T G C G C A A C G C G T
XmnI  (2311)
1 site
G A A N N N N T T C C T T N N N N A A G
SspI  (2106)
1 site
A A T A T T T T A T A A
BglII  (4177)
1 site
A G A T C T T C T A G A
HindIII  (127)
1 site
A A G C T T T T C G A A
AsiSI  (186)
1 site
G C G A T C G C C G C T A G C G
PvuI  (186)
1 site
C G A T C G G C T A G C
SgfI  (186)
1 site
G C G A T C G C C G C T A G C G
BtgI  (189)
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  (189)
1 site
C C A T G G G G T A C C
Eco53kI  (570)
1 site
G A G C T C C T C G A G
EcoICRI  (570)
1 site
G A G C T C C T C G A G
SacI  (572)
1 site
G A G C T C C T C G A G
PaeR7I  (573)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (573)
1 site
C T C G A G G A G C T C
PflMI  (659)
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  (717)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (735)
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  (737)
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  (867)
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  (867)
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  (869)
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  (899)
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  (1068)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BclI  (1099)
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  (1105)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1105)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PshAI  (1164)
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  (1251)
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  (1450)
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  (1476)
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  (1518)
1 site
T C T A G A A G A T C T
SalI  (1524)
1 site
G T C G A C C A G C T G
PstI  (1534)
1 site
C T G C A G G A C G T C
SbfI  (1534)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (1537)
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  (1537)
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  (1599)
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  (1629)
1 site
G C G G C C G C C G C C G G C G
AanI  (1752)
1 site
T T A T A A A A T A T T
PsiI  (1752)
1 site
T T A T A A A A T A T T
HpaI  (1772)
1 site
G T T A A C C A A T T G
MfeI  (1781)
1 site
C A A T T G G T T A A C
SnaBI  (1973)
1 site
T A C G T A A T G C A T
TEV site
588 .. 608  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
TEV site
588 .. 608  =  21 bp
7 amino acids  =  900.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
HaloTag®
624 .. 1505  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
HaloTag®
624 .. 1505  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
AmpR
2124 .. 2984  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2124 .. 2192  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2124 .. 2984  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2193 .. 2984  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2124 .. 2984  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3142 .. 3730  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3142 .. 3730  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
213 .. 548  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
213 .. 548  =  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
3846 .. 4129  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3846 .. 4129  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
SV40 poly(A) signal
1651 .. 1772  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1651 .. 1772  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2019 .. 2123  =  105 bp
AmpR promoter
2019 .. 2123  =  105 bp
CMVd1 promoter
1 .. 100  =  100 bp
truncated human cytomegalovirus (CMV) immediate early promoter that yields slightly reduced expression relative to the full-length promoter
CMVd1 promoter
1 .. 100  =  100 bp
truncated human cytomegalovirus (CMV) immediate early promoter that yields slightly reduced expression relative to the full-length promoter
T7 promoter
132 .. 150  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
132 .. 150  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
156 .. 174  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
156 .. 174  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
ORF:  1877 .. 2227  =  351 bp
ORF:  116 amino acids  =  13.1 kDa
ORF:  4091 .. 178  =  270 bp
ORF:  89 amino acids  =  10.2 kDa
ORF:  213 .. 548  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  555 .. 1511  =  957 bp
ORF:  318 amino acids  =  36.0 kDa
ORF:  2124 .. 2984  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  552 .. 1181  =  630 bp
ORF:  209 amino acids  =  23.0 kDa
ORF:  1181 .. 1417  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  2588 .. 2854  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Download pFC15A HaloTag CMVd1.dna file

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