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Plasmid Files

pEX-N-His

PrecisionShuttle™ bacterial vector for inducible expression of a protein with a cleavable N-terminal 6xHis tag.

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pEX-N-His Sequence and MappEX-N-His.dna
Map and Sequence File   
Sequence Author:  OriGene
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 EcoRI (179) AanI - PsiI (4467) DraIII (4342) BsaAI (4339) XmnI (3789) ScaI (3670) TatI (3668) BsaI (3251) AhdI (3190) AlwNI (2713) T7 promoter lac operator RBS ATG NsiI (292) 6xHis TEV Site AsiSI - SgfI (334) MreI - SgrAI (336) AscI (340) BglII (347) HindIII (354) NheI (365) BmtI (369) RsrII (371) MluI (377) NotI (389) AvaI - BsoBI - PaeR7I - PspXI - XhoI (395) BmeT110I (396) PmeI (404) FseI (414) BlpI (444) StyI (466) EcoO109I (471) BtgI (522) SacII (525) PflMI (757) BstAPI (858) BclI * (1189) BstEII (1356) PspOMI (1382) ApaI (1386) EcoRV (1625) HincII - HpaI (1681) BsmBI (1790) KasI (1814) NarI * (1815) SfoI (1816) PluTI (1818) BamHI (2168) pEX-N-His 4539 bp
EcoRI  (179)
1 site
G A A T T C C T T A A G
AanI  (4467)
1 site
T T A T A A A A T A T T
PsiI  (4467)
1 site
T T A T A A A A T A T T
DraIII  (4342)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BsaAI  (4339)
1 site
Y A C G T R R T G C A Y
XmnI  (3789)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3670)
1 site
A G T A C T T C A T G A
TatI  (3668)
1 site
W G T A C W W C A T G W
BsaI  (3251)
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.
AhdI  (3190)
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.
AlwNI  (2713)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
NsiI  (292)
1 site
A T G C A T T A C G T A
AsiSI  (334)
1 site
G C G A T C G C C G C T A G C G
SgfI  (334)
1 site
G C G A T C G C C G C T A G C G
MreI  (336)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (336)
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.
AscI  (340)
1 site
G G C G C G C C C C G C G C G G
BglII  (347)
1 site
A G A T C T T C T A G A
HindIII  (354)
1 site
A A G C T T T T C G A A
NheI  (365)
1 site
G C T A G C C G A T C G
BmtI  (369)
1 site
G C T A G C C G A T C G
RsrII  (371)
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.
MluI  (377)
1 site
A C G C G T T G C G C A
NotI  (389)
1 site
G C G G C C G C C G C C G G C G
AvaI  (395)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (395)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
PaeR7I  (395)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (395)
1 site
V C T C G A G B B G A G C T C V
XhoI  (395)
1 site
C T C G A G G A G C T C
BmeT110I  (396)
1 site
C Y C G R G G R G C Y C
PmeI  (404)
1 site
G T T T A A A C C A A A T T T G
FseI  (414)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
BlpI  (444)
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.
StyI  (466)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
EcoO109I  (471)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
BtgI  (522)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SacII  (525)
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.
PflMI  (757)
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.
BstAPI  (858)
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.
BclI  (1189)
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.
BstEII  (1356)
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.
PspOMI  (1382)
1 site
G G G C C C C C C G G G
ApaI  (1386)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoRV  (1625)
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.
HincII  (1681)
1 site
G T Y R A C C A R Y T G
HpaI  (1681)
1 site
G T T A A C C A A T T G
BsmBI  (1790)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
KasI  (1814)
1 site
G G C G C C C C G C G G
NarI  (1815)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (1816)
1 site
G G C G C C C C G C G G
PluTI  (1818)
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.
BamHI  (2168)
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.
lacI
825 .. 1907  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
825 .. 1907  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
AmpR
3117 .. 3977  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3117 .. 3908  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3117 .. 3977  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3909 .. 3977  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3117 .. 3977  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2358 .. 2946  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2358 .. 2946  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
4109 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4109 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
AmpR promoter
3978 .. 4082  =  105 bp
AmpR promoter
3978 .. 4082  =  105 bp
MCS
330 .. 416  =  87 bp
multiple cloning site
MCS
330 .. 416  =  87 bp
multiple cloning site
lacI promoter
747 .. 824  =  78 bp
lacI promoter
747 .. 824  =  78 bp
T7 terminator
455 .. 501  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
455 .. 501  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac operator
230 .. 254  =  25 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
230 .. 254  =  25 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
TEV Site
309 .. 329  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV Site
309 .. 329  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
T7 promoter
211 .. 229  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
211 .. 229  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
291 .. 308  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
291 .. 308  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
274 .. 279  =  6 bp
ribosome binding site
RBS
274 .. 279  =  6 bp
ribosome binding site
ATG
288 .. 290  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
288 .. 290  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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