YEp357R

Yeast episomal vector with a URA3 marker for construction of lacZ fusions. The MCS is reversed in YEp357.
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HindIII (1) AhdI (6905) AseI (6730) BsaAI - SnaBI (5208) BstAPI (4486) SphI (11) PstI - SbfI (17) SalI (19) XbaI (25) BamHI (31) TspMI - XmaI (36) SmaI (38) Acc65I (40) KpnI (44) EcoRI (52) Bsu36I (285) BspDI - ClaI (885) BsaBI * (1392) BclI * (1407) BssHII - MauBI (1559) BsiWI (2832) BlpI (3074) PsiI (3303) StuI (3682) PspOMI * (3740) ApaI * (3744) BsmI (3758) BstBI (3851) NcoI (3909) PpuMI (3962) YEp357R 7970 bp
HindIII  (1)
1 site
A A G C T T T T C G A A
AhdI  (6905)
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.
AseI  (6730)
1 site
A T T A A T T A A T T A
BsaAI  (5208)
1 site
Y A C G T R R T G C A Y
SnaBI  (5208)
1 site
T A C G T A A T G C A T
BstAPI  (4486)
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.
SphI  (11)
1 site
G C A T G C C G T A C G
PstI  (17)
1 site
C T G C A G G A C G T C
SbfI  (17)
1 site
C C T G C A G G G G A C G T C C
SalI  (19)
1 site
G T C G A C C A G C T G
XbaI  (25)
1 site
T C T A G A A G A T C T
BamHI  (31)
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.
TspMI  (36)
1 site
C C C G G G G G G C C C
XmaI  (36)
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.
SmaI  (38)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (40)
1 site
G G T A C C C C A T G G
KpnI  (44)
1 site
G G T A C C C C A T G G
EcoRI  (52)
1 site
G A A T T C C T T A A G
Bsu36I  (285)
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.
BspDI  (885)
1 site
A T C G A T T A G C T A
ClaI  (885)
1 site
A T C G A T T A G C T A
BsaBI  (1392)
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  (1407)
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  (1559)
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  (1559)
1 site
C G C G C G C G G C G C G C G C
BsiWI  (2832)
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.
BlpI  (3074)
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.
PsiI  (3303)
1 site
T T A T A A A A T A T T
StuI  (3682)
1 site
A G G C C T T C C G G A
PspOMI  (3740)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI  (3744)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI can be used between 25°C and 37°C.
BsmI  (3758)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
BstBI  (3851)
1 site
T T C G A A A A G C T T
NcoI  (3909)
1 site
C C A T G G G G T A C C
PpuMI  (3962)
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.
lacZ
74 .. 3121  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
lacZ
74 .. 3121  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
2μ ori
4567 .. 5731  =  1165 bp
yeast 2μ plasmid origin of replication
2μ ori
4567 .. 5731  =  1165 bp
yeast 2μ plasmid origin of replication
AmpR
6118 .. 6978  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6118 .. 6186  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6118 .. 6978  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6187 .. 6978  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6118 .. 6978  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
URA3
3316 .. 4119  =  804 bp
267 amino acids  =  29.3 kDa
Product: orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis
yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)
URA3
3316 .. 4119  =  804 bp
267 amino acids  =  29.3 kDa
Product: orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis
yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)
ori
7149 .. 7737  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
7149 .. 7737  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
URA3 promoter
4120 .. 4325  =  206 bp
URA3 promoter
4120 .. 4325  =  206 bp
AmpR promoter
6013 .. 6117  =  105 bp
AmpR promoter
6013 .. 6117  =  105 bp
MCS
1 .. 57  =  57 bp
pUC19 multiple cloning site
MCS
1 .. 57  =  57 bp
pUC19 multiple cloning site
ORF:  2995 .. 3243  =  249 bp
ORF:  82 amino acids  =  9.6 kDa
ORF:  4081 .. 4305  =  225 bp
ORF:  74 amino acids  =  8.7 kDa
ORF:  6118 .. 6978  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  323 .. 3121  =  2799 bp
ORF:  932 amino acids  =  105.9 kDa
ORF:  3608 .. 4084  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
ORF:  4602 .. 4892  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
ORF:  6582 .. 6848  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  920 .. 1234  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2345 .. 2581  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  1984 .. 2295  =  312 bp
ORF:  103 amino acids  =  10.4 kDa
ORF:  3316 .. 4119  =  804 bp
ORF:  267 amino acids  =  29.3 kDa
ORF:  4348 .. 4587  =  240 bp
ORF:  79 amino acids  =  9.8 kDa
ORF:  5368 .. 5634  =  267 bp
ORF:  88 amino acids  =  10.3 kDa
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Download YEp357R.dna file

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Individual Sequences & Maps

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