YEplac181

Yeast episomal plasmid with a LEU2 marker. See also YEplac112 and YEplac195.
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BspQI - SapI (5732) DrdI (5513) PspFI (5315) BseYI (5311) AlwNI (5206) AhdI (4727) BsaI (4661) NmeAIII (4580) ScaI (4246) AatII (3808) ZraI (3806) SacII (3519) PpuMI (3429) PflMI (3413) BstEII (3282) AflII (3002) lac operator HindIII (233) SphI (243) PstI - SbfI (249) SalI (251) AccI (252) XbaI (257) BamHI (263) TspMI - XmaI (268) SmaI (270) Acc65I (272) KpnI (276) Eco53kI (280) BanII - SacI (282) EcoRI (284) KasI (445) NarI (446) SfoI (447) PluTI (449) PfoI * (640) BstAPI (662) HpaI (742) NsiI (1032) BsaAI - SnaBI (1384) BseRI (2024) BsrGI (2196) EcoRV (2571) BstXI (2610) YEplac181 5741 bp
BspQI  (5732)
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  (5732)
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.
DrdI  (5513)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PspFI  (5315)
1 site
C C C A G C G G G T C G
BseYI  (5311)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AlwNI  (5206)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AhdI  (4727)
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  (4661)
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.
NmeAIII  (4580)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
ScaI  (4246)
1 site
A G T A C T T C A T G A
AatII  (3808)
1 site
G A C G T C C T G C A G
ZraI  (3806)
1 site
G A C G T C C T G C A G
SacII  (3519)
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.
PpuMI  (3429)
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.
PflMI  (3413)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BstEII  (3282)
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.
AflII  (3002)
1 site
C T T A A G G A A T T C
HindIII  (233)
1 site
A A G C T T T T C G A A
SphI  (243)
1 site
G C A T G C C G T A C G
PstI  (249)
1 site
C T G C A G G A C G T C
SbfI  (249)
1 site
C C T G C A G G G G A C G T C C
SalI  (251)
1 site
G T C G A C C A G C T G
AccI  (252)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
XbaI  (257)
1 site
T C T A G A A G A T C T
BamHI  (263)
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  (268)
1 site
C C C G G G G G G C C C
XmaI  (268)
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  (270)
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  (272)
1 site
G G T A C C C C A T G G
KpnI  (276)
1 site
G G T A C C C C A T G G
Eco53kI  (280)
1 site
G A G C T C C T C G A G
BanII  (282)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (282)
1 site
G A G C T C C T C G A G
EcoRI  (284)
1 site
G A A T T C C T T A A G
KasI  (445)
1 site
G G C G C C C C G C G G
NarI  (446)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (447)
1 site
G G C G C C C C G C G G
PluTI  (449)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
PfoI  (640)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
BstAPI  (662)
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.
HpaI  (742)
1 site
G T T A A C C A A T T G
NsiI  (1032)
1 site
A T G C A T T A C G T A
BsaAI  (1384)
1 site
Y A C G T R R T G C A Y
SnaBI  (1384)
1 site
T A C G T A A T G C A T
BseRI  (2024)
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  (2196)
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.
EcoRV  (2571)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
BstXI  (2610)
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.
2μ ori
745 .. 2091  =  1347 bp
yeast 2μ plasmid origin of replication
2μ ori
745 .. 2091  =  1347 bp
yeast 2μ plasmid origin of replication
LEU2
2227 .. 3321  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
LEU2
2227 .. 3321  =  1095 bp
364 amino acids  =  39.0 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
AmpR
3940 .. 4800  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3940 .. 4008  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3940 .. 4800  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4009 .. 4800  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3940 .. 4800  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
4971 .. 5559  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
4971 .. 5559  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
LEU2 promoter
3322 .. 3729  =  408 bp
LEU2 promoter
3322 .. 3729  =  408 bp
lacZα
216 .. 500  =  285 bp
95 amino acids  =  11.0 kDa
Product: LacZα fragment of β-galactosidase
lacZα
216 .. 500  =  285 bp
95 amino acids  =  11.0 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
3835 .. 3939  =  105 bp
AmpR promoter
3835 .. 3939  =  105 bp
lac promoter
142 .. 172  =  31 bp
3 segments
   Segment 1:  -35  
   142 .. 147  =  6 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
   Segment 2:  
   148 .. 165  =  18 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
   Segment 3:  -10  
   166 .. 172  =  7 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
180 .. 196  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
180 .. 196  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
MCS
233 .. 289  =  57 bp
pUC19 multiple cloning site
MCS
233 .. 289  =  57 bp
pUC19 multiple cloning site
M13 rev
204 .. 220  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
204 .. 220  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
290 .. 306  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
290 .. 306  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  778 .. 1068  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
ORF:  3940 .. 4800  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2348 .. 2659  =  312 bp
ORF:  103 amino acids  =  11.1 kDa
ORF:  216 .. 530  =  315 bp
ORF:  104 amino acids  =  12.1 kDa
ORF:  4404 .. 4670  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  143 .. 499  =  357 bp
ORF:  118 amino acids  =  13.5 kDa
ORF:  524 .. 763  =  240 bp
ORF:  79 amino acids  =  9.8 kDa
ORF:  1544 .. 1810  =  267 bp
ORF:  88 amino acids  =  10.3 kDa
ORF:  2227 .. 3321  =  1095 bp
ORF:  364 amino acids  =  39.0 kDa
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Download YEplac181.dna file

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