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Plasmid Files

pEX-N-GST

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

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pEX-N-GST Sequence and MappEX-N-GST.dna
Map and Sequence File   
Sequence Author:  OriGene
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 AanI - PsiI (5128) DraIII (5003) BsaAI (5000) BsaI (3912) AhdI (3851) AlwNI (3374) BamHI (2829) PluTI (2479) EcoRI (186) T7 promoter EcoNI (305) MscI (502) BstBI (692) SwaI (722) TEV Site AsiSI - SgfI (995) MreI - SgrAI (997) AscI (1001) BglII (1008) HindIII (1015) NheI (1026) BmtI (1030) RsrII (1032) MluI (1038) NotI (1050) AvaI - BsoBI - PaeR7I - PspXI - XhoI (1056) BmeT110I (1057) PmeI (1065) FseI (1075) BlpI (1105) StyI (1127) BtgI (1183) SacII (1186) PflMI (1418) BstAPI (1519) BstEII (2017) PspOMI (2043) ApaI (2047) HincII - HpaI (2342) BsmBI (2451) KasI (2475) NarI * (2476) SfoI (2477) pEX-N-GST 5200 bp
AanI  (5128)
1 site
T T A T A A A A T A T T
PsiI  (5128)
1 site
T T A T A A A A T A T T
DraIII  (5003)
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  (5000)
1 site
Y A C G T R R T G C A Y
BsaI  (3912)
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  (3851)
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  (3374)
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.
BamHI  (2829)
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.
PluTI  (2479)
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.
EcoRI  (186)
1 site
G A A T T C C T T A A G
EcoNI  (305)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
MscI  (502)
1 site
T G G C C A A C C G G T
BstBI  (692)
1 site
T T C G A A A A G C T T
SwaI  (722)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
AsiSI  (995)
1 site
G C G A T C G C C G C T A G C G
SgfI  (995)
1 site
G C G A T C G C C G C T A G C G
MreI  (997)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (997)
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  (1001)
1 site
G G C G C G C C C C G C G C G G
BglII  (1008)
1 site
A G A T C T T C T A G A
HindIII  (1015)
1 site
A A G C T T T T C G A A
NheI  (1026)
1 site
G C T A G C C G A T C G
BmtI  (1030)
1 site
G C T A G C C G A T C G
RsrII  (1032)
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  (1038)
1 site
A C G C G T T G C G C A
NotI  (1050)
1 site
G C G G C C G C C G C C G G C G
AvaI  (1056)
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  (1056)
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  (1056)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1056)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1056)
1 site
C T C G A G G A G C T C
BmeT110I  (1057)
1 site
C Y C G R G G R G C Y C
PmeI  (1065)
1 site
G T T T A A A C C A A A T T T G
FseI  (1075)
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  (1105)
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  (1127)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BtgI  (1183)
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  (1186)
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  (1418)
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  (1519)
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.
BstEII  (2017)
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  (2043)
1 site
G G G C C C C C C G G G
ApaI  (2047)
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  (2342)
1 site
G T Y R A C C A R Y T G
HpaI  (2342)
1 site
G T T A A C C A A T T G
BsmBI  (2451)
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  (2475)
1 site
G G C G C C C C G C G G
NarI  (2476)
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  (2477)
1 site
G G C G C C C C G C G G
lacI
1486 .. 2568  =  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
1486 .. 2568  =  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
3778 .. 4638  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3778 .. 4569  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3778 .. 4638  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4570 .. 4638  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3778 .. 4638  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
GST
295 .. 948  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from
Schistosoma japonicum
GST
295 .. 948  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from
Schistosoma japonicum
ori
3019 .. 3607  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3019 .. 3607  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
4770 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4770 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
AmpR promoter
4639 .. 4743  =  105 bp
AmpR promoter
4639 .. 4743  =  105 bp
MCS
991 .. 1077  =  87 bp
multiple cloning site
MCS
991 .. 1077  =  87 bp
multiple cloning site
lacI promoter
1408 .. 1485  =  78 bp
lacI promoter
1408 .. 1485  =  78 bp
T7 terminator
1116 .. 1162  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
1116 .. 1162  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac operator
237 .. 261  =  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
237 .. 261  =  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
970 .. 990  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV Site
970 .. 990  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
T7 promoter
218 .. 236  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
218 .. 236  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
281 .. 286  =  6 bp
ribosome binding site
RBS
281 .. 286  =  6 bp
ribosome binding site
stop codon
1044 .. 1046  =  3 bp
stop codon
1044 .. 1046  =  3 bp
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