pFC15K HaloTag CMVd1

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

Sequence Author: Promega

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ScaI (4137) MluI (4053) BspEI (3840) BspHI (3796) ApaLI (3390) PciI (3076) AgeI (3008) BstBI (2981) RsrII (2815) NaeI (2801) NgoMIV (2799) FspI (2401) XcmI (2115) HindIII (127) AsiSI - PvuI - SgfI (186) BtgI - NcoI (189) AhdI (562) Eco53kI - EcoICRI (570) SacI (572) PaeR7I - XhoI (573) PflMI (659) AleI (717) BstEII (735) BseRI (737) BmgBI (869) PasI (899) BsaI (1050) StuI * (1068) BspDI * - ClaI * (1105) AclI (1114) PshAI (1164) BglI (1223) SgrAI (1251) BbsI (1450) BssHII (1476) XbaI (1518) SalI (1524) SbfI (1534) BlpI (1599) NotI (1629) AanI - AanI - PsiI (1752) HpaI (1772) MfeI (1781) BstAPI (1997) pFC15K HaloTag® CMVd1 4177 bp
ScaI  (4137)
1 site
A G T A C T T C A T G A
MluI  (4053)
1 site
A C G C G T T G C G C A
BspEI  (3840)
1 site
T C C G G A A G G C C T
BspHI  (3796)
1 site
T C A T G A A G T A C T
ApaLI  (3390)
1 site
G T G C A C C A C G T G
PciI  (3076)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (3008)
1 site
A C C G G T T G G C C A
BstBI  (2981)
1 site
T T C G A A A A G C T T
RsrII  (2815)
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  (2801)
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  (2799)
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.
FspI  (2401)
1 site
T G C G C A A C G C G T
XcmI  (2115)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
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
AhdI  (562)
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.
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.
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.
BsaI  (1050)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
StuI  (1068)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
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.
AclI  (1114)
1 site
A A C G T T T T G C A A
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.
BglI  (1223)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
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
SbfI  (1534)
1 site
C C T G C A G G G G A C G T C C
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
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
BstAPI  (1997)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
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
NeoR/KanR
2171 .. 2965  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2171 .. 2965  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3137 .. 3725  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3137 .. 3725  =  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
3841 .. 4124  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3841 .. 4124  =  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
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:  2171 .. 2965  =  795 bp
ORF:  264 amino acids  =  29.0 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:  2343 .. 2729  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  4086 .. 178  =  270 bp
ORF:  89 amino acids  =  10.2 kDa
ORF:  1181 .. 1417  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  2480 .. 2734  =  255 bp
ORF:  84 amino acids  =  9.8 kDa
ORF:  552 .. 1181  =  630 bp
ORF:  209 amino acids  =  23.0 kDa
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Download pFC15K HaloTag CMVd1.dna file

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