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Plasmid Files

YEp364

Yeast episomal vector with a LEU2 marker for construction of lacZ fusions. For other reading frames, use YEp363 or YEp365.

 
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 SmaI (8) TspMI - XmaI (6) BspQI - SapI (8344) AhdI (7339) BsaI (7273) ScaI (6858) BsaAI - SnaBI (5824) NsiI (5472) PpuMI (4883) BstEII (4738) KpnI (4530) Acc65I (4526) AgeI (4523) BamHI (11) SalI (17) PstI (27) HindIII (31) Bsu36I (257) Bpu10I (538) DraIII (1223) BsaBI * (1364) BclI * (1379) BssHII - MauBI (1531) Eco53kI (1971) SacI (1973) BstZ17I (2798) BsiWI (2804) NdeI (2990) BlpI (3046) BseRI (3433) BsrGI (3653) AflII (4458) YEp364 8404 bp
SmaI  (8)
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  (6)
1 site
C C C G G G G G G C C C
XmaI  (6)
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.
BspQI  (8344)
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  (8344)
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.
AhdI  (7339)
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.
BsaI  (7273)
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.
ScaI  (6858)
1 site
A G T A C T T C A T G A
BsaAI  (5824)
1 site
Y A C G T R R T G C A Y
SnaBI  (5824)
1 site
T A C G T A A T G C A T
NsiI  (5472)
1 site
A T G C A T T A C G T A
PpuMI  (4883)
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.
BstEII  (4738)
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.
KpnI  (4530)
1 site
G G T A C C C C A T G G
Acc65I  (4526)
1 site
G G T A C C C C A T G G
AgeI  (4523)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
BamHI  (11)
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.
SalI  (17)
1 site
G T C G A C C A G C T G
PstI  (27)
1 site
C T G C A G G A C G T C
HindIII  (31)
1 site
A A G C T T T T C G A A
Bsu36I  (257)
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  (538)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
DraIII  (1223)
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.
BsaBI  (1364)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BclI  (1379)
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  (1531)
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  (1531)
1 site
C G C G C G C G G C G C G C G C
Eco53kI  (1971)
1 site
G A G C T C C T C G A G
SacI  (1973)
1 site
G A G C T C C T C G A G
BstZ17I  (2798)
1 site
G T A T A C C A T A T G
BsiWI  (2804)
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.
NdeI  (2990)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
BlpI  (3046)
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.
BseRI  (3433)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
BsrGI  (3653)
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.
AflII  (4458)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
lacZ
46 .. 3093  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
lacZ
46 .. 3093  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
LEU2
3683 .. 4777  =  1095 bp
364 amino acids  =  39.2 kDa
Product: 3-isopropylmalate dehydrogenase, required
for leucine biosynthesis
yeast auxotrophic marker
LEU2
3683 .. 4777  =  1095 bp
364 amino acids  =  39.2 kDa
Product: 3-isopropylmalate dehydrogenase, required
for leucine biosynthesis
yeast auxotrophic marker
2μ ori
5183 .. 6165  =  983 bp
yeast 2μ plasmid origin of replication
2μ ori
5183 .. 6165  =  983 bp
yeast 2μ plasmid origin of replication
AmpR
6552 .. 7412  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6552 .. 6620  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6552 .. 7412  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6621 .. 7412  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6552 .. 7412  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
7583 .. 8171  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
7583 .. 8171  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
LEU2 promoter
4778 .. 5182  =  405 bp
LEU2 promoter
4778 .. 5182  =  405 bp
AmpR promoter
6447 .. 6551  =  105 bp
AmpR promoter
6447 .. 6551  =  105 bp
MCS
1 .. 36  =  36 bp
multiple cloning site
MCS
1 .. 36  =  36 bp
multiple cloning site
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