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Plasmid Files

pGEX-1λT

Bacterial vector for expressing GST fusion proteins. The frame of the EcoRI site matches that of lambda gt11.

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pGEX-1_lambda_T Sequence and MappGEX-1_lambda_T.dna
Map and Sequence File   
Sequence Author:  GE Healthcare
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 BtgI (4847) Bsu36I (4739) PluTI (4289) SfoI (4287) NarI * (4286) KasI (4285) HpaI (4152) EcoRV (4096) BssHII (4057) ApaI - BanII (3857) PspOMI (3853) BstEII (3827) MluI (3646) BstAPI (3329) PflMI (3228) AlwNI (2621) BfuAI - BspMI (62) lac operator EcoNI (268) MscI (465) BstBI (655) SwaI (685) pGEX 5' Sequencing Primer (869 .. 891) BamHI (930) EcoRI (939) stop codons PfoI (1014) pGEX 3' Sequencing Primer (997 .. 1019) PflFI - Tth111I (1117) BsaAI (1124) ZraI (1221) AatII (1223) PstI (1900) BsaI (2076) AhdI (2142) pGEX-1 λ T 4947 bp
BtgI  (4847)
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  (4739)
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  (4289)
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  (4287)
1 site
G G C G C C C C G C G G
NarI  (4286)
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  (4285)
1 site
G G C G C C C C G C G G
HpaI  (4152)
1 site
G T T A A C C A A T T G
EcoRV  (4096)
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  (4057)
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  (3857)
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  (3857)
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  (3853)
1 site
G G G C C C C C C G G G
BstEII  (3827)
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  (3646)
1 site
A C G C G T T G C G C A
BstAPI  (3329)
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  (3228)
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  (2621)
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  (930)
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  (939)
1 site
G A A T T C C T T A A G
PfoI  (1014)
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  (1117)
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  (1117)
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  (1124)
1 site
Y A C G T R R T G C A Y
ZraI  (1221)
1 site
G A C G T C C T G C A G
AatII  (1223)
1 site
G A C G T C C T G C A G
PstI  (1900)
1 site
C T G C A G G A C G T C
BsaI  (2076)
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  (2142)
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
997 .. 1019  =  23 annealed bases
Tm  =  66°C
lacI
3296 .. 4378  =  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
3296 .. 4378  =  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
1355 .. 2215  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   1355 .. 1423  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1355 .. 2215  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   1424 .. 2215  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1355 .. 2215  =  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
2386 .. 2974  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2386 .. 2974  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
1250 .. 1354  =  105 bp
AmpR promoter
1250 .. 1354  =  105 bp
lacIq promoter
3218 .. 3295  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
lacIq promoter
3218 .. 3295  =  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
thrombin site
918 .. 935  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
918 .. 935  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
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
949 .. 959  =  11 bp
stop codons in all three reading frames
stop codons
949 .. 959  =  11 bp
stop codons in all three reading frames
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