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Plasmid Files

pGEX-3X

Bacterial vector for expressing GST fusions with a Factor Xa site.

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pGEX-3X Sequence and MappGEX-3X.dna
Map and Sequence File   
Sequence Author:  GE Healthcare
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 BtgI (4852) Bsu36I (4744) PluTI (4294) SfoI (4292) NarI * (4291) KasI (4290) HpaI (4157) EcoRV (4101) BssHII (4062) ApaI - BanII (3862) PspOMI (3858) BstEII (3832) MluI (3651) BstAPI (3334) PflMI (3233) AlwNI (2626) BfuAI - BspMI (62) lac operator EcoNI (268) MscI (465) BstBI (655) SwaI (685) pGEX 5' Sequencing Primer (869 .. 891) Factor Xa site BamHI (934) AvaI - BsoBI - TspMI - XmaI (939) BmeT110I (940) SmaI (941) EcoRI (944) stop codons PfoI (1019) pGEX 3' Sequencing Primer (1002 .. 1024) PflFI - Tth111I (1122) BsaAI (1129) ZraI (1226) AatII (1228) PstI (1905) BsaI (2081) AhdI (2147) pGEX-3X 4952 bp
BtgI  (4852)
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  (4744)
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  (4294)
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  (4292)
1 site
G G C G C C C C G C G G
NarI  (4291)
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  (4290)
1 site
G G C G C C C C G C G G
HpaI  (4157)
1 site
G T T A A C C A A T T G
EcoRV  (4101)
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  (4062)
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.
ApaI  (3862)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (3862)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PspOMI  (3858)
1 site
G G G C C C C C C G G G
BstEII  (3832)
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  (3651)
1 site
A C G C G T T G C G C A
BstAPI  (3334)
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  (3233)
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  (2626)
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.
BamHI  (934)
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.
AvaI  (939)
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  (939)
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.
TspMI  (939)
1 site
C C C G G G G G G C C C
XmaI  (939)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
BmeT110I  (940)
1 site
C Y C G R G G R G C Y C
SmaI  (941)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (944)
1 site
G A A T T C C T T A A G
PfoI  (1019)
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  (1122)
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  (1122)
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  (1129)
1 site
Y A C G T R R T G C A Y
ZraI  (1226)
1 site
G A C G T C C T G C A G
AatII  (1228)
1 site
G A C G T C C T G C A G
PstI  (1905)
1 site
C T G C A G G A C G T C
BsaI  (2081)
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  (2147)
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  =  67°C
pGEX 3' Sequencing Primer
23-mer  /  65% GC
1 binding site
1002 .. 1024  =  23 annealed bases
Tm  =  66°C
lacI
3301 .. 4383  =  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
3301 .. 4383  =  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
1360 .. 2220  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   1360 .. 1428  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1360 .. 2220  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   1429 .. 2220  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1360 .. 2220  =  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
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
AmpR promoter
1255 .. 1359  =  105 bp
AmpR promoter
1255 .. 1359  =  105 bp
lacIq promoter
3223 .. 3300  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
lacIq promoter
3223 .. 3300  =  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
   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
   Segment 2:  
   189 .. 204  =  16 bp
strong E. coli promoter; hybrid between the trp and
lac UV5 promoters
tac promoter
183 .. 211  =  29 bp
   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).
MCS
934 .. 949  =  16 bp
multiple cloning site
MCS
934 .. 949  =  16 bp
multiple cloning site
Factor Xa site
921 .. 932  =  12 bp
4 amino acids  =  473.5 Da
Product: Factor Xa recognition and cleavage site
Factor Xa site
921 .. 932  =  12 bp
4 amino acids  =  473.5 Da
Product: Factor Xa recognition and cleavage site
stop codons
954 .. 964  =  11 bp
stop codons in all three reading frames
stop codons
954 .. 964  =  11 bp
stop codons in all three reading frames
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