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Plasmid Files

pFN24K HaloTag® CMVd3

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

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pFN24K HaloTag CMVd3 Sequence and MappFN24K HaloTag CMVd3.dna
Map and Sequence File   
Sequence Author:  Promega
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 ScaI (4088) MluI (4004) BspEI (3791) BspHI (3747) ApaLI (3341) PciI (3027) AgeI (2959) BstBI (2932) RsrII (2766) NaeI (2752) NgoMIV (2750) FspI (2352) XcmI (2066) HindIII (72) PflMI (177) AleI (235) BstEII (253) BseRI (255) BpmI (308) BmgBI (387) PasI (417) BsaI (568) StuI * (586) BspDI * - ClaI * (623) AclI (632) PshAI (682) BglI (741) SgrAI (769) BbsI (968) BssHII (994) PaeR7I - XhoI (1012) AsiSI - PvuI - SgfI (1070) BtgI - NcoI (1073) DraI - PmeI (1437) Acc65I (1454) KpnI (1458) XbaI (1469) SbfI (1485) BlpI (1550) NotI (1580) AanI - AanI - PsiI (1703) HpaI (1723) MfeI (1732) BstAPI (1948) pFN24K HaloTag® CMVd3 4128 bp
ScaI  (4088)
1 site
A G T A C T T C A T G A
MluI  (4004)
1 site
A C G C G T T G C G C A
BspEI  (3791)
1 site
T C C G G A A G G C C T
BspHI  (3747)
1 site
T C A T G A A G T A C T
ApaLI  (3341)
1 site
G T G C A C C A C G T G
PciI  (3027)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (2959)
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  (2932)
1 site
T T C G A A A A G C T T
RsrII  (2766)
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  (2752)
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  (2750)
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  (2352)
1 site
T G C G C A A C G C G T
XcmI  (2066)
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
PflMI  (177)
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  (235)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (253)
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  (255)
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  (308)
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  (387)
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  (417)
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  (568)
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  (586)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BspDI  (623)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (623)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AclI  (632)
1 site
A A C G T T T T G C A A
PshAI  (682)
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  (741)
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  (769)
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  (968)
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  (994)
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  (1012)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1012)
1 site
C T C G A G G A G C T C
AsiSI  (1070)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1070)
1 site
C G A T C G G C T A G C
SgfI  (1070)
1 site
G C G A T C G C C G C T A G C G
BtgI  (1073)
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  (1073)
1 site
C C A T G G G G T A C C
DraI  (1437)
1 site
T T T A A A A A A T T T
PmeI  (1437)
1 site
G T T T A A A C C A A A T T T G
Acc65I  (1454)
1 site
G G T A C C C C A T G G
KpnI  (1458)
1 site
G G T A C C C C A T G G
XbaI  (1469)
1 site
T C T A G A A G A T C T
SbfI  (1485)
1 site
C C T G C A G G G G A C G T C C
BlpI  (1550)
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  (1580)
1 site
G C G G C C G C C G C C G G C G
AanI  (1703)
1 site
T T A T A A A A T A T T
AanI  (1703)
1 site
T T A T A A A A T A T T
PsiI  (1703)
1 site
T T A T A A A A T A T T
HpaI  (1723)
1 site
G T T A A C C A A T T G
MfeI  (1732)
1 site
C A A T T G G T T A A C
BstAPI  (1948)
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®
136 .. 1026  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
HaloTag®
136 .. 1026  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
NeoR/KanR
2122 .. 2916  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
2122 .. 2916  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
3088 .. 3676  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3088 .. 3676  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
barnase
1097 .. 1432  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus
amyloliquefaciens

The barnase gene is lethal in standard bacterial
transformation strains.
barnase
1097 .. 1432  =  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
3792 .. 4075  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
cer region
3792 .. 4075  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
SV40 poly(A) signal
1602 .. 1723  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1602 .. 1723  =  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
1039 .. 1059  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV protease recognition site
TEV site
1039 .. 1059  =  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
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