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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.dna
Map and Sequence File:    Download    Open   
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) EcoNI (305) MscI (502) BstBI (692) SwaI (722) 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
ORF:  295 .. 1026  =  732 bp
ORF:  243 amino acids  =  28.1 kDa
ORF:  1609 .. 2568  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  3908 .. 4174  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1419 .. 1736  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  2325 .. 2588  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  2691 .. 2918  =  228 bp
ORF:  75 amino acids  =  8.5 kDa
ORF:  2351 .. 2602  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  3778 .. 4638  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  732 .. 959  =  228 bp
ORF:  75 amino acids  =  8.3 kDa
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