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Plasmid Files

YEp356

Yeast episomal vector with a URA3 marker for construction of lacZ fusions. The MCS is reversed in YEp356R.

 
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 EcoRI (1) AhdI (6900) AseI (6725) BsaAI - SnaBI (5203) BstAPI (4481) PpuMI (3957) Acc65I (13) KpnI - TspMI - XmaI (17) SmaI (19) BamHI (22) XbaI (28) SalI (34) PstI - SbfI (44) SphI (50) HindIII (52) Bsu36I (280) BspDI - ClaI (880) BsaBI * (1387) BclI * (1402) BssHII - MauBI (1554) BsiWI (2827) BlpI (3069) PsiI (3298) StuI (3677) PspOMI * (3735) ApaI * (3739) BsmI (3753) BstBI (3846) NcoI (3904) YEp356 7965 bp
EcoRI  (1)
1 site
G A A T T C C T T A A G
AhdI  (6900)
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  (6725)
1 site
A T T A A T T A A T T A
BsaAI  (5203)
1 site
Y A C G T R R T G C A Y
SnaBI  (5203)
1 site
T A C G T A A T G C A T
BstAPI  (4481)
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  (3957)
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.
Acc65I  (13)
1 site
G G T A C C C C A T G G
KpnI  (17)
1 site
G G T A C C C C A T G G
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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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.
BspDI  (880)
1 site
A T C G A T T A G C T A
ClaI  (880)
1 site
A T C G A T T A G C T A
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
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.
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.
PsiI  (3298)
1 site
T T A T A A A A T A T T
StuI  (3677)
1 site
A G G C C T T C C G G A
PspOMI  (3735)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI  (3739)
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  (3753)
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  (3846)
1 site
T T C G A A A A G C T T
NcoI  (3904)
1 site
C C A T G G G G T A C 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
2μ ori
4562 .. 5726  =  1165 bp
yeast 2μ plasmid origin of replication
2μ ori
4562 .. 5726  =  1165 bp
yeast 2μ plasmid origin of replication
AmpR
6113 .. 6973  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6113 .. 6181  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6113 .. 6973  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6182 .. 6973  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6113 .. 6973  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
URA3
3311 .. 4114  =  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
3311 .. 4114  =  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
7144 .. 7732  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
7144 .. 7732  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
URA3 promoter
4115 .. 4320  =  206 bp
URA3 promoter
4115 .. 4320  =  206 bp
AmpR promoter
6008 .. 6112  =  105 bp
AmpR promoter
6008 .. 6112  =  105 bp
MCS
1 .. 57  =  57 bp
polylinker
MCS
1 .. 57  =  57 bp
polylinker
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