YEp366

Yeast episomal vector with a LEU2 marker for construction of lacZ fusions. The MCS is reversed in YEp366R.
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TspMI - XmaI (17) BspQI - SapI (8367) AhdI (7362) BsaI (7296) ScaI (6881) BsaAI - SnaBI (5847) NsiI (5495) PpuMI (4906) BstEII (4761) AgeI (4546) SmaI (19) BamHI (22) XbaI (28) SalI (34) PstI - SbfI (44) SphI (50) HindIII (52) Bsu36I (280) Bpu10I (561) DraIII (1246) BsaBI * (1387) BclI * (1402) BssHII - MauBI (1554) BstZ17I (2821) BsiWI (2827) NdeI (3013) BlpI (3069) BseRI (3456) BsrGI (3676) AflII (4481) YEp366 8427 bp
TspMI  (17)
1 site
C C C G G G G G G C C C
XmaI  (17)
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.
BspQI  (8367)
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  (8367)
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  (7362)
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  (7296)
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  (6881)
1 site
A G T A C T T C A T G A
BsaAI  (5847)
1 site
Y A C G T R R T G C A Y
SnaBI  (5847)
1 site
T A C G T A A T G C A T
NsiI  (5495)
1 site
A T G C A T T A C G T A
PpuMI  (4906)
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  (4761)
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.
AgeI  (4546)
1 site
A C C G G T T G G C C A
SmaI  (19)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (22)
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.
XbaI  (28)
1 site
T C T A G A A G A T C T
SalI  (34)
1 site
G T C G A C C A G C T G
PstI  (44)
1 site
C T G C A G G A C G T C
SbfI  (44)
1 site
C C T G C A G G G G A C G T C C
SphI  (50)
1 site
G C A T G C C G T A C G
HindIII  (52)
1 site
A A G C T T T T C G A A
Bsu36I  (280)
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  (561)
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.
DraIII  (1246)
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  (1387)
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  (1402)
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  (1554)
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  (1554)
1 site
C G C G C G C G G C G C G C G C
BstZ17I  (2821)
1 site
G T A T A C C A T A T G
BsiWI  (2827)
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  (3013)
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  (3069)
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  (3456)
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  (3676)
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  (4481)
1 site
C T T A A G G A A T T C
lacZ
69 .. 3116  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
lacZ
69 .. 3116  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
LEU2
3706 .. 4800  =  1095 bp
364 amino acids  =  39.2 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
LEU2
3706 .. 4800  =  1095 bp
364 amino acids  =  39.2 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
2μ ori
5206 .. 6188  =  983 bp
yeast 2μ plasmid origin of replication
2μ ori
5206 .. 6188  =  983 bp
yeast 2μ plasmid origin of replication
AmpR
6575 .. 7435  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6575 .. 6643  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6575 .. 7435  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6644 .. 7435  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6575 .. 7435  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
7606 .. 8194  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
7606 .. 8194  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
LEU2 promoter
4801 .. 5205  =  405 bp
LEU2 promoter
4801 .. 5205  =  405 bp
AmpR promoter
6470 .. 6574  =  105 bp
AmpR promoter
6470 .. 6574  =  105 bp
MCS
1 .. 57  =  57 bp
polylinker
MCS
1 .. 57  =  57 bp
polylinker
ORF:  2990 .. 3250  =  261 bp
ORF:  86 amino acids  =  10.1 kDa
ORF:  6575 .. 7435  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  48 .. 3116  =  3069 bp
ORF:  1022 amino acids  =  116.2 kDa
ORF:  5241 .. 5531  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
ORF:  3706 .. 4800  =  1095 bp
ORF:  364 amino acids  =  39.2 kDa
ORF:  7039 .. 7305  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  915 .. 1229  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2340 .. 2576  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  1979 .. 2290  =  312 bp
ORF:  103 amino acids  =  10.4 kDa
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Download YEp366.dna file

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Individual Sequences & Maps

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