YEp354

Yeast episomal vector with a URA3 marker for construction of lacZ fusions. For other reading frames, use YEp353 or YEp355.
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EcoRI (1) AhdI (6877) AseI (6702) BsaAI - SnaBI (5180) BstAPI (4458) PpuMI (3934) TspMI - XmaI (6) SmaI (8) BamHI (11) SalI (17) PstI (27) HindIII (31) Bsu36I (257) BspDI - ClaI (857) BsaBI * (1364) BclI * (1379) BssHII - MauBI (1531) Eco53kI (1971) SacI (1973) BsiWI (2804) BlpI (3046) PsiI (3275) StuI (3654) PspOMI * (3712) ApaI * (3716) BsmI (3730) BstBI (3823) NcoI (3881) YEp354 7942 bp
EcoRI  (1)
1 site
G A A T T C C T T A A G
AhdI  (6877)
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  (6702)
1 site
A T T A A T T A A T T A
BsaAI  (5180)
1 site
Y A C G T R R T G C A Y
SnaBI  (5180)
1 site
T A C G T A A T G C A T
BstAPI  (4458)
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.
PpuMI  (3934)
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.
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

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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.
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.
BspDI  (857)
1 site
A T C G A T T A G C T A
ClaI  (857)
1 site
A T C G A T T A G C T A
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
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.
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.
PsiI  (3275)
1 site
T T A T A A A A T A T T
StuI  (3654)
1 site
A G G C C T T C C G G A
PspOMI  (3712)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI  (3716)
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  (3730)
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  (3823)
1 site
T T C G A A A A G C T T
NcoI  (3881)
1 site
C C A T G G G G T A C C
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
2μ ori
4539 .. 5703  =  1165 bp
yeast 2μ plasmid origin of replication
2μ ori
4539 .. 5703  =  1165 bp
yeast 2μ plasmid origin of replication
AmpR
6090 .. 6950  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6090 .. 6158  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6090 .. 6950  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6159 .. 6950  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6090 .. 6950  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
URA3
3288 .. 4091  =  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
3288 .. 4091  =  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
7121 .. 7709  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
7121 .. 7709  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
URA3 promoter
4092 .. 4297  =  206 bp
URA3 promoter
4092 .. 4297  =  206 bp
AmpR promoter
5985 .. 6089  =  105 bp
AmpR promoter
5985 .. 6089  =  105 bp
MCS
1 .. 36  =  36 bp
multiple cloning site
MCS
1 .. 36  =  36 bp
multiple cloning site
ORF:  295 .. 3093  =  2799 bp
ORF:  932 amino acids  =  105.9 kDa
ORF:  3580 .. 4056  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
ORF:  4574 .. 4864  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
ORF:  2967 .. 3215  =  249 bp
ORF:  82 amino acids  =  9.6 kDa
ORF:  4053 .. 4277  =  225 bp
ORF:  74 amino acids  =  8.7 kDa
ORF:  6090 .. 6950  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  6554 .. 6820  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  892 .. 1206  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2317 .. 2553  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  1956 .. 2267  =  312 bp
ORF:  103 amino acids  =  10.4 kDa
ORF:  3288 .. 4091  =  804 bp
ORF:  267 amino acids  =  29.3 kDa
ORF:  4320 .. 4559  =  240 bp
ORF:  79 amino acids  =  9.8 kDa
ORF:  5340 .. 5606  =  267 bp
ORF:  88 amino acids  =  10.3 kDa
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Download YEp354.dna file

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Individual Sequences & Maps

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