pEX2

Vector for bacterial expression of lacZ fusion proteins. For other reading frames, use pEX1 or pEX3.
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BspEI * (124) EcoO109I (5762) ScaI (5266) BsaI (4847) AhdI (4786) BspQI - SapI (3777) BstAPI (3717) BsaAI (3647) PflFI - Tth111I (3640) XbaI (3260) PstI (3222) SalI (3212) BamHI (3206) SmaI (3203) TspMI - XmaI (3201) EcoRI (3196) BsiWI (2963) KasI (174) NarI * (175) SfoI (176) PluTI (178) Bsu36I (416) Bpu10I (697) EcoRV (1305) DraIII (1382) BclI * (1538) BssHII - MauBI (1690) AfeI (2027) Eco53kI (2130) SacI (2132) pEX2 5787 bp
BspEI  (124)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
EcoO109I  (5762)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
ScaI  (5266)
1 site
A G T A C T T C A T G A
BsaI  (4847)
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  (4786)
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.
BspQI  (3777)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (3777)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
BstAPI  (3717)
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.
BsaAI  (3647)
1 site
Y A C G T R R T G C A Y
PflFI  (3640)
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  (3640)
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.
XbaI  (3260)
1 site
T C T A G A A G A T C T
PstI  (3222)
1 site
C T G C A G G A C G T C
SalI  (3212)
1 site
G T C G A C C A G C T G
BamHI  (3206)
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.
SmaI  (3203)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (3201)
1 site
C C C G G G G G G C C C
XmaI  (3201)
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.
EcoRI  (3196)
1 site
G A A T T C C T T A A G
BsiWI  (2963)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
KasI  (174)
1 site
G G C G C C C C G C G G
NarI  (175)
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  (176)
1 site
G G C G C C C C G C G G
PluTI  (178)
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.
Bsu36I  (416)
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.
Bpu10I  (697)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
EcoRV  (1305)
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.
DraIII  (1382)
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.
BclI  (1538)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BssHII  (1690)
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.
MauBI  (1690)
1 site
C G C G C G C G G C G C G C G C
AfeI  (2027)
1 site
A G C G C T T C G C G A
Eco53kI  (2130)
1 site
G A G C T C C T C G A G
SacI  (2132)
1 site
G A G C T C C T C G A G
cro-lacZ
85 .. 3240  =  3156 bp
1051 amino acids  =  119.2 kDa
4 segments
   Segment 1:  cro  
   85 .. 108  =  24 bp
   8 amino acids  =  1.0 kDa
Product: Cro-β-galactosidase hybrid protein
cro-lacZ
85 .. 3240  =  3156 bp
1051 amino acids  =  119.2 kDa
4 segments
   Segment 2:  
   109 .. 249  =  141 bp
   47 amino acids  =  5.2 kDa
Product: Cro-β-galactosidase hybrid protein
cro-lacZ
85 .. 3240  =  3156 bp
1051 amino acids  =  119.2 kDa
4 segments
   Segment 3:  lacZ  
   250 .. 3195  =  2946 bp
   982 amino acids  =  111.5 kDa
Product: Cro-β-galactosidase hybrid protein
cro-lacZ
85 .. 3240  =  3156 bp
1051 amino acids  =  119.2 kDa
4 segments
   Segment 4:  
   3196 .. 3240  =  45 bp
   14 amino acids  =  1.5 kDa
Product: Cro-β-galactosidase hybrid protein
cro-lacZ
85 .. 3240  =  3156 bp
1051 amino acids  =  119.2 kDa
4 segments
Product: Cro-β-galactosidase hybrid protein
AmpR
4713 .. 5573  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4713 .. 5504  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4713 .. 5573  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5505 .. 5573  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4713 .. 5573  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3954 .. 4542  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3954 .. 4542  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
5574 .. 5678  =  105 bp
AmpR promoter
5574 .. 5678  =  105 bp
ORF:  85 .. 3240  =  3156 bp
ORF:  1051 amino acids  =  119.2 kDa
ORF:  4843 .. 5109  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1051 .. 1365  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2476 .. 2712  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  2115 .. 2426  =  312 bp
ORF:  103 amino acids  =  10.4 kDa
ORF:  4713 .. 5573  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pEX2.dna file

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