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Plasmid Files

pHTN HaloTag® CMV-neo

Mammalian expression vector with a traditional MCS and ampicillin and G418 resistance markers, encoding a cleavable N-terminal HaloTag®.

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pHTN HaloTag CMV-neo Sequence and MappHTN HaloTag CMV-neo.dna
Map and Sequence File   
Sequence Author:  Promega
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 BglII (6138) MluI (6019) BspEI (5806) PciI (5042) AgeI (4974) AhdI (4872) XmnI (4272) BamHI (3840) BstXI (3767) RsrII (3562) NaeI (3548) NgoMIV (3546) PluTI (3049) SfoI (3047) BsrGI (96) SpeI (152) NdeI (387) T7 promoter NheI (1052) BmtI (1056) PflMI (1108) AleI (1166) BstEII (1184) BmgBI (1318) PasI (1348) BclI * (1548) PshAI (1613) SgrAI (1700) TspMI - XmaI (1870) SmaI (1872) SalI (1936) AccI (1937) PaeR7I - XhoI (1943) TEV site AsiSI - PvuI (2001) EcoRI (2009) Eco53kI (2018) SacI (2020) SacII (2027) EcoRV (2031) XbaI (2033) PspOMI (2042) ApaI (2046) SbfI (2057) NotI (2066) BlpI (2097) PsiI (2245) HpaI (2265) MfeI (2274) Acc65I (2502) KpnI (2506) SexAI * (2619) SfiI (2805) AvrII (2852) KasI (3045) NarI (3046) pHTN HaloTag® CMV-neo 6143 bp
BglII  (6138)
1 site
A G A T C T T C T A G A
MluI  (6019)
1 site
A C G C G T T G C G C A
BspEI  (5806)
1 site
T C C G G A A G G C C T
PciI  (5042)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (4974)
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.
AhdI  (4872)
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.
XmnI  (4272)
1 site
G A A N N N N T T C C T T N N N N A A G
BamHI  (3840)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
BstXI  (3767)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
RsrII  (3562)
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  (3548)
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  (3546)
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.
PluTI  (3049)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (3047)
1 site
G G C G C C C C G C G G
BsrGI  (96)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SpeI  (152)
1 site
A C T A G T T G A T C A
NdeI  (387)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
NheI  (1052)
1 site
G C T A G C C G A T C G
BmtI  (1056)
1 site
G C T A G C C G A T C G
PflMI  (1108)
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  (1166)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (1184)
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.
BmgBI  (1318)
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  (1348)
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.
BclI  (1548)
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.
PshAI  (1613)
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  (1700)
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.
TspMI  (1870)
1 site
C C C G G G G G G C C C
XmaI  (1870)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (1872)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SalI  (1936)
1 site
G T C G A C C A G C T G
AccI  (1937)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
PaeR7I  (1943)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1943)
1 site
C T C G A G G A G C T C
AsiSI  (2001)
1 site
G C G A T C G C C G C T A G C G
PvuI  (2001)
1 site
C G A T C G G C T A G C
EcoRI  (2009)
1 site
G A A T T C C T T A A G
Eco53kI  (2018)
1 site
G A G C T C C T C G A G
SacI  (2020)
1 site
G A G C T C C T C G A G
SacII  (2027)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
EcoRV  (2031)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
XbaI  (2033)
1 site
T C T A G A A G A T C T
PspOMI  (2042)
1 site
G G G C C C C C C G G G
ApaI  (2046)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SbfI  (2057)
1 site
C C T G C A G G G G A C G T C C
NotI  (2066)
1 site
G C G G C C G C C G C C G G C G
BlpI  (2097)
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.
PsiI  (2245)
1 site
T T A T A A A A T A T T
HpaI  (2265)
1 site
G T T A A C C A A T T G
MfeI  (2274)
1 site
C A A T T G G T T A A C
Acc65I  (2502)
1 site
G G T A C C C C A T G G
KpnI  (2506)
1 site
G G T A C C C C A T G G
SexAI  (2619)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (2805)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AvrII  (2852)
1 site
C C T A G G G G A T C C
KasI  (3045)
1 site
G G C G C C C C G C G G
NarI  (3046)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
HaloTag®
1067 .. 1957  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
HaloTag®
1067 .. 1957  =  891 bp
297 amino acids  =  33.6 kDa
Product: modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives
AmpR
4085 .. 4945  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4085 .. 4153  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4085 .. 4945  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4154 .. 4945  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4085 .. 4945  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
2918 .. 3712  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
2918 .. 3712  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
5103 .. 5691  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5103 .. 5691  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
138 .. 517  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
2510 .. 2867  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2510 .. 2867  =  358 bp
SV40 enhancer and early promoter
cer region
5807 .. 6090  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
cer region
5807 .. 6090  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
2144 .. 2265  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2144 .. 2265  =  122 bp
SV40 polyadenylation signal
AmpR promoter
3980 .. 4084  =  105 bp
AmpR promoter
3980 .. 4084  =  105 bp
MCS
1997 .. 2072  =  76 bp
multiple cloning site
MCS
1997 .. 2072  =  76 bp
multiple cloning site
poly(A) signal
3776 .. 3824  =  49 bp
synthetic polyadenylation signal
poly(A) signal
3776 .. 3824  =  49 bp
synthetic polyadenylation signal
TEV site
1970 .. 1990  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV protease recognition site
TEV site
1970 .. 1990  =  21 bp
7 amino acids  =  901.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV protease recognition site
T7 promoter
1033 .. 1051  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1033 .. 1051  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
2718 .. 2853  =  136 bp
SV40 origin of replication
SV40 ori
2718 .. 2853  =  136 bp
SV40 origin of replication
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