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pFC17K HaloTag® CMVd3

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

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pFC17K HaloTag CMVd3.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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ScaI (4082) MluI (3998) BspEI (3785) BspHI (3741) ApaLI (3335) PciI (3021) AgeI (2953) BstBI (2926) RsrII (2760) NaeI (2746) NgoMIV (2744) FspI (2346) XcmI (2060) HindIII (72) AsiSI - PvuI - SgfI (131) BtgI - NcoI (134) AhdI (507) Eco53kI - EcoICRI (515) SacI (517) PaeR7I - XhoI (518) PflMI (604) AleI (662) BstEII (680) BseRI (682) BmgBI (814) PasI (844) BsaI (995) StuI * (1013) BspDI * - ClaI * (1050) AclI (1059) PshAI (1109) BglI (1168) SgrAI (1196) BbsI (1395) BssHII (1421) XbaI (1463) SalI (1469) SbfI (1479) BlpI (1544) NotI (1574) AanI - AanI - PsiI (1697) HpaI (1717) MfeI (1726) BstAPI (1942) pFC17K HaloTag® CMVd3 4122 bp
ScaI  (4082)
1 site
A G T A C T T C A T G A
MluI  (3998)
1 site
A C G C G T T G C G C A
BspEI  (3785)
1 site
T C C G G A A G G C C T
BspHI  (3741)
1 site
T C A T G A A G T A C T
ApaLI  (3335)
1 site
G T G C A C C A C G T G
PciI  (3021)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (2953)
1 site
A C C G G T T G G C C A
BstBI  (2926)
1 site
T T C G A A A A G C T T
RsrII  (2760)
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  (2746)
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  (2744)
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  (2346)
1 site
T G C G C A A C G C G T
XcmI  (2060)
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  (72)
1 site
A A G C T T T T C G A A
AsiSI  (131)
1 site
G C G A T C G C C G C T A G C G
PvuI  (131)
1 site
C G A T C G G C T A G C
SgfI  (131)
1 site
G C G A T C G C C G C T A G C G
BtgI  (134)
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  (134)
1 site
C C A T G G G G T A C C
AhdI  (507)
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  (515)
1 site
G A G C T C C T C G A G
EcoICRI  (515)
1 site
G A G C T C C T C G A G
SacI  (517)
1 site
G A G C T C C T C G A G
PaeR7I  (518)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (518)
1 site
C T C G A G G A G C T C
PflMI  (604)
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  (662)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (680)
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  (682)
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  (814)
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  (844)
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  (995)
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  (1013)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BspDI  (1050)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1050)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AclI  (1059)
1 site
A A C G T T T T G C A A
PshAI  (1109)
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  (1168)
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  (1196)
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  (1395)
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  (1421)
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  (1463)
1 site
T C T A G A A G A T C T
SalI  (1469)
1 site
G T C G A C C A G C T G
SbfI  (1479)
1 site
C C T G C A G G G G A C G T C C
BlpI  (1544)
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  (1574)
1 site
G C G G C C G C C G C C G G C G
AanI  (1697)
1 site
T T A T A A A A T A T T
AanI  (1697)
1 site
T T A T A A A A T A T T
PsiI  (1697)
1 site
T T A T A A A A T A T T
HpaI  (1717)
1 site
G T T A A C C A A T T G
MfeI  (1726)
1 site
C A A T T G G T T A A C
BstAPI  (1942)
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.
HaloTag®
569 .. 1450  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
HaloTag®
569 .. 1450  =  882 bp
294 amino acids  =  33.4 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
NeoR/KanR
2116 .. 2910  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2116 .. 2910  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3082 .. 3670  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3082 .. 3670  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
158 .. 493  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
158 .. 493  =  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
3786 .. 4069  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3786 .. 4069  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
SV40 poly(A) signal
1596 .. 1717  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1596 .. 1717  =  122 bp
SV40 polyadenylation signal
CMVd3 promoter
1 .. 45  =  45 bp
truncated human cytomegalovirus (CMV) immediate early promoter that yields strongly reduced expression relative to the full-length promoter
CMVd3 promoter
1 .. 45  =  45 bp
truncated human cytomegalovirus (CMV) immediate early promoter that yields strongly reduced expression relative to the full-length promoter
TEV site
533 .. 553  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
TEV site
533 .. 553  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
T7 promoter
77 .. 95  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
77 .. 95  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
101 .. 119  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
101 .. 119  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
ORF:  2116 .. 2910  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  158 .. 493  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  500 .. 1456  =  957 bp
ORF:  318 amino acids  =  36.0 kDa
ORF:  2288 .. 2674  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  1126 .. 1362  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  2425 .. 2679  =  255 bp
ORF:  84 amino acids  =  9.8 kDa
ORF:  497 .. 1126  =  630 bp
ORF:  209 amino acids  =  23.0 kDa
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