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pFN22K HaloTag® CMVd1

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

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pFN22K HaloTag CMVd1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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ScaI (4143) MluI (4059) BspEI (3846) BspHI (3802) ApaLI (3396) PciI (3082) AgeI (3014) BstBI (2987) RsrII (2821) NaeI (2807) NgoMIV (2805) FspI (2407) XcmI (2121) HindIII (127) PflMI (232) AleI (290) BstEII (308) BseRI (310) BpmI (363) BmgBI (442) PasI (472) BsaI (623) StuI * (641) BspDI * - ClaI * (678) AclI (687) PshAI (737) BglI (796) SgrAI (824) BbsI (1023) BssHII (1049) PaeR7I - XhoI (1067) AsiSI - PvuI - SgfI (1125) BtgI - NcoI (1128) DraI - PmeI (1492) Acc65I (1509) KpnI (1513) XbaI (1524) SbfI (1540) BlpI (1605) NotI (1635) AanI - AanI - PsiI (1758) HpaI (1778) MfeI (1787) BstAPI (2003) pFN22K HaloTag® CMVd1 4183 bp
ScaI  (4143)
1 site
A G T A C T T C A T G A
MluI  (4059)
1 site
A C G C G T T G C G C A
BspEI  (3846)
1 site
T C C G G A A G G C C T
BspHI  (3802)
1 site
T C A T G A A G T A C T
ApaLI  (3396)
1 site
G T G C A C C A C G T G
PciI  (3082)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (3014)
1 site
A C C G G T T G G C C A
BstBI  (2987)
1 site
T T C G A A A A G C T T
RsrII  (2821)
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  (2807)
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  (2805)
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  (2407)
1 site
T G C G C A A C G C G T
XcmI  (2121)
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
PflMI  (232)
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  (290)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (308)
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  (310)
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.
BpmI  (363)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BmgBI  (442)
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  (472)
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  (623)
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  (641)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BspDI  (678)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (678)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AclI  (687)
1 site
A A C G T T T T G C A A
PshAI  (737)
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  (796)
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  (824)
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  (1023)
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  (1049)
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.
PaeR7I  (1067)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1067)
1 site
C T C G A G G A G C T C
AsiSI  (1125)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1125)
1 site
C G A T C G G C T A G C
SgfI  (1125)
1 site
G C G A T C G C C G C T A G C G
BtgI  (1128)
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  (1128)
1 site
C C A T G G G G T A C C
DraI  (1492)
1 site
T T T A A A A A A T T T
PmeI  (1492)
1 site
G T T T A A A C C A A A T T T G
Acc65I  (1509)
1 site
G G T A C C C C A T G G
KpnI  (1513)
1 site
G G T A C C C C A T G G
XbaI  (1524)
1 site
T C T A G A A G A T C T
SbfI  (1540)
1 site
C C T G C A G G G G A C G T C C
BlpI  (1605)
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  (1635)
1 site
G C G G C C G C C G C C G G C G
AanI  (1758)
1 site
T T A T A A A A T A T T
AanI  (1758)
1 site
T T A T A A A A T A T T
PsiI  (1758)
1 site
T T A T A A A A T A T T
HpaI  (1778)
1 site
G T T A A C C A A T T G
MfeI  (1787)
1 site
C A A T T G G T T A A C
BstAPI  (2003)
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®
191 .. 1081  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
HaloTag®
191 .. 1081  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives
NeoR/KanR
2177 .. 2971  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2177 .. 2971  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3143 .. 3731  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3143 .. 3731  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
1152 .. 1487  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
1152 .. 1487  =  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
3847 .. 4130  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3847 .. 4130  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
SV40 poly(A) signal
1657 .. 1778  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1657 .. 1778  =  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
TEV site
1094 .. 1114  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
TEV site
1094 .. 1114  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV protease recognition site
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:  191 .. 1138  =  948 bp
ORF:  315 amino acids  =  35.7 kDa
ORF:  2177 .. 2971  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1152 .. 1487  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  2349 .. 2735  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  4092 .. 178  =  270 bp
ORF:  89 amino acids  =  10.2 kDa
ORF:  158 .. 1153  =  996 bp
ORF:  331 amino acids  =  37.5 kDa
ORF:  2486 .. 2740  =  255 bp
ORF:  84 amino acids  =  9.8 kDa
ORF:  754 .. 990  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  969 .. 1208  =  240 bp
ORF:  79 amino acids  =  8.9 kDa
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