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Plasmid Files

YEp13

Yeast episomal vector with a LEU2 marker.

 
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YEp13.dna
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NheI (232) XbaI (9966) AvrII (8451) AleI (8196) BstXI (6557) AgeI (6092) BstEII (5876) SacII (5646) BmtI (236) BamHI (378) SgrAI (413) SphI (569) PshAI (719) EagI (942) BsmI (1362) BsgI * (1638) BspEI * (1667) PvuII (2071) BspQI - SapI (2362) AhdI (3371) ZraI (4291) AatII (4293) StuI (4447) BlpI (4481) DraIII (4673) AbsI - PaeR7I - PspXI - XhoI (5194) YEp13 10,667 bp
NheI  (232)
1 site
G C T A G C C G A T C G
XbaI  (9966)
1 site
T C T A G A A G A T C T
AvrII  (8451)
1 site
C C T A G G G G A T C C
AleI  (8196)
1 site
C A C N N N N G T G G T G N N N N C A C
BstXI  (6557)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
AgeI  (6092)
1 site
A C C G G T T G G C C A
BstEII  (5876)
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.
SacII  (5646)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
BmtI  (236)
1 site
G C T A G C C G A T C G
BamHI  (378)
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.
SgrAI  (413)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
SphI  (569)
1 site
G C A T G C C G T A C G
PshAI  (719)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
EagI  (942)
1 site
C G G C C G G C C G G C
BsmI  (1362)
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.
BsgI  (1638)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
* Blocked by EcoKI methylation.
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BspEI  (1667)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
PvuII  (2071)
1 site
C A G C T G G T C G A C
BspQI  (2362)
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  (2362)
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  (3371)
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.
ZraI  (4291)
1 site
G A C G T C C T G C A G
AatII  (4293)
1 site
G A C G T C C T G C A G
StuI  (4447)
1 site
A G G C C T T C C G G A
BlpI  (4481)
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.
DraIII  (4673)
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.
AbsI  (5194)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (5194)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (5194)
1 site
V C T C G A G B B G A G C T C V
XhoI  (5194)
1 site
C T C G A G G A G C T C
2μ ori
8988 .. 10,330  =  1343 bp
yeast 2μ plasmid origin of replication
2μ ori
8988 .. 10,330  =  1343 bp
yeast 2μ plasmid origin of replication
TcR
89 .. 1279  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
89 .. 1279  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
LEU2
5843 .. 6937  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
LEU2
5843 .. 6937  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
AmpR
3298 .. 4158  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3298 .. 4089  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3298 .. 4158  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   4090 .. 4158  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3298 .. 4158  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2539 .. 3127  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
2539 .. 3127  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
LEU2 promoter
5435 .. 5842  =  408 bp
LEU2 promoter
5435 .. 5842  =  408 bp
rop
1920 .. 2111  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
1920 .. 2111  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
AmpR promoter
4159 .. 4263  =  105 bp
AmpR promoter
4159 .. 4263  =  105 bp
tet promoter
13 .. 41  =  29 bp
   Segment 1:  -35  
   13 .. 18  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
13 .. 41  =  29 bp
   Segment 2:  
   19 .. 35  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
13 .. 41  =  29 bp
   Segment 3:  -10  
   36 .. 41  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
13 .. 41  =  29 bp
3 segments
E. coli promoter for tetracycline efflux protein gene
ORF:  262 .. 645  =  384 bp
ORF:  127 amino acids  =  14.7 kDa
ORF:  1183 .. 1521  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  89 .. 1279  =  1191 bp
ORF:  396 amino acids  =  41.5 kDa
ORF:  3428 .. 3694  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5843 .. 6937  =  1095 bp
ORF:  364 amino acids  =  39.0 kDa
ORF:  9023 .. 9313  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
ORF:  1821 .. 2111  =  291 bp
ORF:  96 amino acids  =  11.4 kDa
ORF:  4578 .. 4934  =  357 bp
ORF:  118 amino acids  =  13.5 kDa
ORF:  8400 .. 8663  =  264 bp
ORF:  87 amino acids  =  9.8 kDa
ORF:  1518 .. 1916  =  399 bp
ORF:  132 amino acids  =  15.3 kDa
ORF:  7296 .. 8537  =  1242 bp
ORF:  413 amino acids  =  45.8 kDa
ORF:  8769 .. 9008  =  240 bp
ORF:  79 amino acids  =  9.8 kDa
ORF:  9789 .. 10,109  =  321 bp
ORF:  106 amino acids  =  12.9 kDa
ORF:  326 .. 568  =  243 bp
ORF:  80 amino acids  =  8.0 kDa
ORF:  626 .. 1084  =  459 bp
ORF:  152 amino acids  =  16.7 kDa
ORF:  10,151 .. 10,492  =  342 bp
ORF:  113 amino acids  =  13.0 kDa
ORF:  442 .. 792  =  351 bp
ORF:  116 amino acids  =  12.2 kDa
ORF:  3298 .. 4158  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  6505 .. 6816  =  312 bp
ORF:  103 amino acids  =  11.1 kDa
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