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Plasmid Files

pEX-C-His

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

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pEX-C-His Sequence and MappEX-C-His.dna
Map and Sequence File   
Sequence Author:  OriGene
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 EcoRI (175) PsiI (4500) DraIII (4375) BsaAI (4372) XmnI (3822) ScaI (3703) TatI (3701) BsaI (3284) AhdI (3223) AlwNI (2746) T7 promoter lac operator RBS BsaBI * - BseRI - BspDI * - ClaI * (279) AscI (295) BglII (302) HindIII (309) NheI (320) BmtI (324) RsrII (326) MluI (332) TEV Site 6xHis NotI (383) AvaI - BsoBI - PaeR7I - PspXI - XhoI (389) BmeT110I (390) TEV Site 6xHis PmeI (437) FseI (447) BlpI (477) StyI (499) EcoO109I (504) BtgI (555) SacII (558) PflMI (790) BstAPI (891) BclI * (1222) BstEII (1389) PspOMI (1415) ApaI (1419) HincII - HpaI (1714) BsmBI (1823) KasI (1847) NarI * (1848) SfoI (1849) PluTI (1851) BamHI (2201) pEX-C-His 4583 bp
EcoRI  (175)
1 site
G A A T T C C T T A A G
PsiI  (4500)
1 site
T T A T A A A A T A T T
DraIII  (4375)
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  (4372)
1 site
Y A C G T R R T G C A Y
XmnI  (3822)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3703)
1 site
A G T A C T T C A T G A
TatI  (3701)
1 site
W G T A C W W C A T G W
BsaI  (3284)
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  (3223)
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  (2746)
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.
BsaBI  (279)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BseRI  (279)
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.
BspDI  (279)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (279)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AscI  (295)
1 site
G G C G C G C C C C G C G C G G
BglII  (302)
1 site
A G A T C T T C T A G A
HindIII  (309)
1 site
A A G C T T T T C G A A
NheI  (320)
1 site
G C T A G C C G A T C G
BmtI  (324)
1 site
G C T A G C C G A T C G
RsrII  (326)
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  (332)
1 site
A C G C G T T G C G C A
NotI  (383)
1 site
G C G G C C G C C G C C G G C G
AvaI  (389)
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  (389)
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  (389)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (389)
1 site
V C T C G A G B B G A G C T C V
XhoI  (389)
1 site
C T C G A G G A G C T C
BmeT110I  (390)
1 site
C Y C G R G G R G C Y C
PmeI  (437)
1 site
G T T T A A A C C A A A T T T G
FseI  (447)
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  (477)
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  (499)
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  (504)
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  (555)
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  (558)
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  (790)
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  (891)
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  (1222)
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  (1389)
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  (1415)
1 site
G G G C C C C C C G G G
ApaI  (1419)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
HincII  (1714)
1 site
G T Y R A C C A R Y T G
HpaI  (1714)
1 site
G T T A A C C A A T T G
BsmBI  (1823)
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  (1847)
1 site
G G C G C C C C G C G G
NarI  (1848)
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  (1849)
1 site
G G C G C C C C G C G G
PluTI  (1851)
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  (2201)
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
858 .. 1940  =  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
858 .. 1940  =  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
3150 .. 4010  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3150 .. 3941  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3150 .. 4010  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3942 .. 4010  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3150 .. 4010  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2391 .. 2979  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2391 .. 2979  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
4142 .. 14  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4142 .. 14  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
AmpR promoter
4011 .. 4115  =  105 bp
AmpR promoter
4011 .. 4115  =  105 bp
lacI promoter
780 .. 857  =  78 bp
lacI promoter
780 .. 857  =  78 bp
MCS
277 .. 337  =  61 bp
multiple cloning site
MCS
277 .. 337  =  61 bp
multiple cloning site
T7 terminator
488 .. 534  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
488 .. 534  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac operator
226 .. 250  =  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
226 .. 250  =  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
338 .. 358  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV Site
338 .. 358  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV Site
395 .. 415  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV Site
395 .. 415  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
T7 promoter
207 .. 225  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
207 .. 225  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
359 .. 376  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
359 .. 376  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
416 .. 433  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
416 .. 433  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
270 .. 275  =  6 bp
ribosome binding site
RBS
270 .. 275  =  6 bp
ribosome binding site
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