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Plasmid Files

pYES2.1-E

High-copy episomal Saccharomyces cerevisiae expression vector that serves as an acceptor in the Echo™ Cloning System.

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pYES2.1-E Sequence and MappYES2.1-E.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 SpeI (5820) NaeI (5680) NgoMIV (5678) SwaI (5352) SnaBI (5048) BmeT110I (4699) AvaI - BsoBI (4698) HpaI (4403) BmgBI (4343) PfoI * (4301) NruI (3997) BlpI (3961) BspDI - ClaI (3817) BmtI (3812) NheI (3808) NdeI (3668) BsgI (3639) SbfI (3605) BfuAI - BspMI (3594) XcmI (3456) EcoRV (3395) NcoI (3374) BsmI (3223) ApaI * (3209) PspOMI * (3205) AgeI (71) PvuII (471) HindIII (501) PmeI (544) BsrGI (736) MluI (743) BciVI (1208) BspHI (1725) AhdI (1898) BglI (2018) BsaHI (2435) BsmBI (2957) pYES2.1-E 5825 bp
SpeI  (5820)
1 site
A C T A G T T G A T C A
NaeI  (5680)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
NgoMIV  (5678)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
SwaI  (5352)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
SnaBI  (5048)
1 site
T A C G T A A T G C A T
BmeT110I  (4699)
1 site
C Y C G R G G R G C Y C
AvaI  (4698)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (4698)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
HpaI  (4403)
1 site
G T T A A C C A A T T G
BmgBI  (4343)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
PfoI  (4301)
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.
NruI  (3997)
1 site
T C G C G A A G C G C T
BlpI  (3961)
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.
BspDI  (3817)
1 site
A T C G A T T A G C T A
ClaI  (3817)
1 site
A T C G A T T A G C T A
BmtI  (3812)
1 site
G C T A G C C G A T C G
NheI  (3808)
1 site
G C T A G C C G A T C G
NdeI  (3668)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
BsgI  (3639)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

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).
SbfI  (3605)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (3594)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (3594)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
XcmI  (3456)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
EcoRV  (3395)
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.
NcoI  (3374)
1 site
C C A T G G G G T A C C
BsmI  (3223)
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.
ApaI  (3209)
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.
PspOMI  (3205)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
AgeI  (71)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
PvuII  (471)
1 site
C A G C T G G T C G A C
HindIII  (501)
1 site
A A G C T T T T C G A A
PmeI  (544)
1 site
G T T T A A A C C A A A T T T G
BsrGI  (736)
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.
MluI  (743)
1 site
A C G C G T T G C G C A
BciVI  (1208)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
BspHI  (1725)
1 site
T C A T G A A G T A C T
AhdI  (1898)
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.
BglI  (2018)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
BsaHI  (2435)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
BsmBI  (2957)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
2μ ori
4404 .. 5284  =  881 bp
yeast 2μ plasmid origin of replication
2μ ori
4404 .. 5284  =  881 bp
yeast 2μ plasmid origin of replication
AmpR
1825 .. 2685  =  861 bp
287 codons
   Segment 2:  
   1825 .. 2616  =  792 bp
   264 codons
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and
related antibiotics
AmpR
1825 .. 2685  =  861 bp
287 codons
   Segment 1:  signal sequence  
   2617 .. 2685  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and
related antibiotics
AmpR
1825 .. 2685  =  861 bp
287 codons
2 segments
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and
related antibiotics
URA3
2781 .. 3584  =  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
2781 .. 3584  =  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
1066 .. 1654  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1066 .. 1654  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
5353 .. 5808  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
5353 .. 5808  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
GAL1 promoter
2 .. 443  =  442 bp
inducible promoter, regulated by Gal4
GAL1 promoter
2 .. 443  =  442 bp
inducible promoter, regulated by Gal4
CYC1 terminator
571 .. 818  =  248 bp
transcription terminator for CYC1
CYC1 terminator
571 .. 818  =  248 bp
transcription terminator for CYC1
URA3 promoter
3585 .. 3800  =  216 bp
URA3 promoter
3585 .. 3800  =  216 bp
loxH
507 .. 540  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT). The loxH variant was
optimized for expression constructs but remains
compatible with loxP.
loxH
507 .. 540  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT). The loxH variant was
optimized for expression constructs but remains
compatible with loxP.
T7 promoter
475 .. 493  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
475 .. 493  =  19 bp
promoter for bacteriophage T7 RNA polymerase
UAS
2 .. 119  =  118 bp
upstream activating sequence mediating
Gal4-dependent induction
UAS
2 .. 119  =  118 bp
upstream activating sequence mediating
Gal4-dependent induction
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