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Plasmid Files

pEG202

Episomal yeast two-hybrid vector for constitutive expression of LexA fused to the N-terminus of a protein. Also known as pLexA.

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pEG202 Sequence and MappEG202.dna
Map and Sequence File   
Sequence Author:  Clontech
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 PflFI - Tth111I (10,062) BspQI - SapI (9923) AatII (7999) ZraI (7997) PpuMI (7854) StuI (7837) BlpI (7800) NruI (7767) SrfI (7604) BstXI (6387) BsiWI (6356) BssHII (5960) BanII - SacI (5117) KasI (180) NarI (181) SfoI (182) PluTI (184) NgoMIV (246) NaeI (248) PacI (721) BsrGI (827) MluI (1616) PmeI (2059) EcoRI (2144) BamHI (2154) MCS NcoI (2165) NotI (2171) PaeR7I - PspXI - XhoI (2177) FspAI (2656) XbaI (3487) AvrII (5004) Eco53kI (5115) pEG202 10,166 bp
PflFI  (10,062)
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  (10,062)
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.
BspQI  (9923)
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  (9923)
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.
AatII  (7999)
1 site
G A C G T C C T G C A G
ZraI  (7997)
1 site
G A C G T C C T G C A G
PpuMI  (7854)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
StuI  (7837)
1 site
A G G C C T T C C G G A
BlpI  (7800)
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.
NruI  (7767)
1 site
T C G C G A A G C G C T
SrfI  (7604)
1 site
G C C C G G G C C G G G C C C G
BstXI  (6387)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BsiWI  (6356)
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.
BssHII  (5960)
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.
BanII  (5117)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (5117)
1 site
G A G C T C C T C G A G
KasI  (180)
1 site
G G C G C C C C G C G G
NarI  (181)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (182)
1 site
G G C G C C C C G C G G
PluTI  (184)
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.
NgoMIV  (246)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (248)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
PacI  (721)
1 site
T T A A T T A A A A T T A A T T
BsrGI  (827)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
MluI  (1616)
1 site
A C G C G T T G C G C A
PmeI  (2059)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (2144)
1 site
G A A T T C C T T A A G
BamHI  (2154)
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.
NcoI  (2165)
1 site
C C A T G G G G T A C C
NotI  (2171)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (2177)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2177)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2177)
1 site
C T C G A G G A G C T C
FspAI  (2656)
1 site
R T G C G C A Y Y A C G C G T R
XbaI  (3487)
1 site
T C T A G A A G A T C T
AvrII  (5004)
1 site
C C T A G G G G A T C C
Eco53kI  (5115)
1 site
G A G C T C C T C G A G
2μ ori
3128 .. 4470  =  1343 bp
yeast 2μ plasmid origin of replication
2μ ori
3128 .. 4470  =  1343 bp
yeast 2μ plasmid origin of replication
AmpR
8131 .. 8991  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   8131 .. 8199  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8131 .. 8991  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   8200 .. 8991  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8131 .. 8991  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
HIS3
6114 .. 6773  =  660 bp
219 amino acids  =  23.8 kDa
Product: imidazoleglycerol-phosphate dehydratase,
required for histidine biosynthesis
yeast auxotrophic marker
HIS3
6114 .. 6773  =  660 bp
219 amino acids  =  23.8 kDa
Product: imidazoleglycerol-phosphate dehydratase,
required for histidine biosynthesis
yeast auxotrophic marker
LexA
1538 .. 2143  =  606 bp
202 amino acids  =  22.4 kDa
   Segment 1:  DNA-binding domain  
   1538 .. 1789  =  252 bp
   84 amino acids  =  9.2 kDa
Product: E. coli DNA-binding protein that represses
genes involved in the SOS response to DNA damage
The N-terminal DNA-binding domain recognizes the
16-bp palindromic sequence
5'-CTGTATATATATACAG-3'.
LexA
1538 .. 2143  =  606 bp
202 amino acids  =  22.4 kDa
   Segment 2:  
   1790 .. 2143  =  354 bp
   118 amino acids  =  13.2 kDa
Product: E. coli DNA-binding protein that represses
genes involved in the SOS response to DNA damage
The N-terminal DNA-binding domain recognizes the
16-bp palindromic sequence
5'-CTGTATATATATACAG-3'.
LexA
1538 .. 2143  =  606 bp
202 amino acids  =  22.4 kDa
2 segments
Product: E. coli DNA-binding protein that represses
genes involved in the SOS response to DNA damage
The N-terminal DNA-binding domain recognizes the
16-bp palindromic sequence
5'-CTGTATATATATACAG-3'.
ori
9162 .. 9750  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
9162 .. 9750  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ADH1 promoter
1110 .. 1511  =  402 bp
promoter for alcohol dehydrogenase 1
ADH1 promoter
1110 .. 1511  =  402 bp
promoter for alcohol dehydrogenase 1
ADH1 terminator
2209 .. 2396  =  188 bp
transcription terminator for alcohol dehydrogenase 1
ADH1 terminator
2209 .. 2396  =  188 bp
transcription terminator for alcohol dehydrogenase 1
HIS3 promoter
6774 .. 6961  =  188 bp
HIS3 promoter
6774 .. 6961  =  188 bp
AmpR promoter
8026 .. 8130  =  105 bp
AmpR promoter
8026 .. 8130  =  105 bp
MCS
2144 .. 2182  =  39 bp
multiple cloning site
MCS
2144 .. 2182  =  39 bp
multiple cloning site
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