pGEX-6P-3

Bacterial vector for expressing GST fusion proteins with a PreScission protease site. For other reading frames, use pGEX-6P-1 or pGEX-6P-2.

Sequence Author: GE Healthcare

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HomePlasmidspGEX Vectors (GE Healthcare)pGEX-6P-3
BtgI (4883) Bsu36I (4775) PluTI (4325) SfoI (4323) NarI * (4322) KasI (4321) EcoRV (4132) BssHII (4093) BstEII (3863) MluI (3682) BstAPI (3365) PflMI (3264) AlwNI (2657) BfuAI - BspMI (62) lac operator EcoNI (268) MscI (465) BstBI (655) SwaI (685) pGEX 5' Sequencing Primer (869 .. 891) PasI (941) BamHI (945) EcoRI (953) TspMI - XmaI (958) SmaI (960) SalI (963) AccI (964) PaeR7I - PspXI - XhoI (968) EagI - NotI (974) PfoI (1050) pGEX 3' Sequencing Primer (1033 .. 1055) PflFI - Tth111I (1153) BsaAI (1160) ZraI (1257) AatII (1259) PstI (1936) BsaI (2112) AhdI (2178) pGEX-6P-3 4983 bp
BtgI  (4883)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
Bsu36I  (4775)
1 site		 C C T N A G G G G A N T C C
Sticky ends from different Bsu36I sites may not be compatible.
PluTI  (4325)
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  (4323)
1 site		 G G C G C C C C G C G G
NarI  (4322)
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.
KasI  (4321)
1 site		 G G C G C C C C G C G G
EcoRV  (4132)
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.
BssHII  (4093)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
BstEII  (3863)
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.
MluI  (3682)
1 site		 A C G C G T T G C G C A
BstAPI  (3365)
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.
PflMI  (3264)
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.
AlwNI  (2657)
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.
BfuAI  (62)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (62)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
EcoNI  (268)
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  (465)
1 site		 T G G C C A A C C G G T
BstBI  (655)
1 site		 T T C G A A A A G C T T
SwaI  (685)
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.
PasI  (941)
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.
BamHI  (945)
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.
EcoRI  (953)
1 site		 G A A T T C C T T A A G
TspMI  (958)
1 site		 C C C G G G G G G C C C
XmaI  (958)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (960)
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  (963)
1 site		 G T C G A C C A G C T G
AccI  (964)
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  (968)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (968)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (968)
1 site		 C T C G A G G A G C T C
EagI  (974)
1 site		 C G G C C G G C C G G C
NotI  (974)
1 site		 G C G G C C G C C G C C G G C G
PfoI  (1050)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
PflFI  (1153)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (1153)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsaAI  (1160)
1 site		 Y A C G T R R T G C A Y
ZraI  (1257)
1 site		 G A C G T C C T G C A G
AatII  (1259)
1 site		 G A C G T C C T G C A G
PstI  (1936)
1 site		 C T G C A G G A C G T C
BsaI  (2112)
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  (2178)
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.
pGEX 5' Sequencing Primer
23-mer  /  65% GC
1 binding site
869 .. 891  =  23 annealed bases
Tm  =  66°C
pGEX 3' Sequencing Primer
23-mer  /  65% GC
1 binding site
1033 .. 1055  =  23 annealed bases
Tm  =  66°C
lacI
3332 .. 4414  =  1083 bp
360 amino acids  =  38.5 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
3332 .. 4414  =  1083 bp
360 amino acids  =  38.5 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
1391 .. 2251  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   1391 .. 1459  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1391 .. 2251  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   1460 .. 2251  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1391 .. 2251  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
GST
258 .. 911  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
258 .. 911  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
PreScission site
918 .. 941  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C and PreScission proteases
PreScission site
918 .. 941  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C and PreScission proteases
ori
2422 .. 3010  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2422 .. 3010  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
1286 .. 1390  =  105 bp
AmpR promoter
1286 .. 1390  =  105 bp
lacIq promoter
3254 .. 3331  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
3254 .. 3331  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
tac promoter
183 .. 211  =  29 bp
3 segments
   Segment 1:  -35  
   183 .. 188  =  6 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
183 .. 211  =  29 bp
3 segments
   Segment 2:  
   189 .. 204  =  16 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
183 .. 211  =  29 bp
3 segments
   Segment 3:  -10  
   205 .. 211  =  7 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
tac promoter
183 .. 211  =  29 bp
3 segments
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
lac operator
219 .. 235  =  17 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
219 .. 235  =  17 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).
stop codons
985 .. 995  =  11 bp
stop codons in all three reading frames
stop codons
985 .. 995  =  11 bp
stop codons in all three reading frames
MCS
940 .. 980  =  41 bp
multiple cloning site
MCS
940 .. 980  =  41 bp
multiple cloning site
ORF:  3265 .. 3582  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  4171 .. 4434  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4537 .. 81  =  528 bp
ORF:  175 amino acids  =  19.7 kDa
ORF:  1391 .. 2251  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  3455 .. 4414  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  4922 .. 178  =  240 bp
ORF:  79 amino acids  =  9.1 kDa
ORF:  258 .. 995  =  738 bp
ORF:  245 amino acids  =  28.3 kDa
ORF:  811 .. 1044  =  234 bp
ORF:  77 amino acids  =  8.4 kDa
ORF:  1855 .. 2121  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4197 .. 4448  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  695 .. 982  =  288 bp
ORF:  95 amino acids  =  10.7 kDa
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Individual Sequences & Maps

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