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Plasmid Files

pYES-DEST52

Gateway® destination vector for galactose-inducible high-level expression of proteins in Saccharomyces cerevisiae.

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pYES-DEST52 Sequence and MappYES-DEST52.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 NaeI (7476) NgoMIV (7474) SwaI (7148) SnaBI (6844) HpaI (6199) PfoI * (6097) NruI (5793) BlpI (5757) BspDI - ClaI (5613) BmtI (5608) NheI (5604) NdeI (5464) BsgI (5435) SbfI (5401) XcmI (5252) EcoRV (5191) BsaHI (4231) BglI (3814) SpeI (7616) T7 promoter SacII (643) BspEI (980) EcoRI (984) PasI - PflMI * (1216) BssHII (1478) BbvCI (1709) TspMI - XmaI (1853) SmaI - SrfI (1855) BsaBI * (1868) BstXI (1972) SalI (2112) 6xHis PmeI (2340) BciVI (3004) BspHI (3521) AhdI (3694) pYES-DEST52 7621 bp
NaeI  (7476)
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  (7474)
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  (7148)
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  (6844)
1 site
T A C G T A A T G C A T
HpaI  (6199)
1 site
G T T A A C C A A T T G
PfoI  (6097)
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  (5793)
1 site
T C G C G A A G C G C T
BlpI  (5757)
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  (5613)
1 site
A T C G A T T A G C T A
ClaI  (5613)
1 site
A T C G A T T A G C T A
BmtI  (5608)
1 site
G C T A G C C G A T C G
NheI  (5604)
1 site
G C T A G C C G A T C G
NdeI  (5464)
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  (5435)
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  (5401)
1 site
C C T G C A G G G G A C G T C C
XcmI  (5252)
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  (5191)
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.
BsaHI  (4231)
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.
BglI  (3814)
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.
SpeI  (7616)
1 site
A C T A G T T G A T C A
SacII  (643)
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.
BspEI  (980)
1 site
T C C G G A A G G C C T
EcoRI  (984)
1 site
G A A T T C C T T A A G
PasI  (1216)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (1216)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BssHII  (1478)
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.
BbvCI  (1709)
1 site
C C T C A G C G G A G T C G
TspMI  (1853)
1 site
C C C G G G G G G C C C
XmaI  (1853)
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  (1855)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (1855)
1 site
G C C C G G G C C G G G C C C G
BsaBI  (1868)
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.
BstXI  (1972)
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.
SalI  (2112)
1 site
G T C G A C C A G C T G
PmeI  (2340)
1 site
G T T T A A A C C A A A T T T G
BciVI  (3004)
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  (3521)
1 site
T C A T G A A G T A C T
AhdI  (3694)
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.
2μ ori
6200 .. 7080  =  881 bp
yeast 2μ plasmid origin of replication
2μ ori
6200 .. 7080  =  881 bp
yeast 2μ plasmid origin of replication
AmpR
3621 .. 4481  =  861 bp
286 amino acids  =  31.7 kDa
   Segment 2:  
   3621 .. 4412  =  792 bp
   263 amino acids  =  29.1 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3621 .. 4481  =  861 bp
286 amino acids  =  31.7 kDa
   Segment 1:  signal sequence  
   4413 .. 4481  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3621 .. 4481  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
URA3
4577 .. 5380  =  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
4577 .. 5380  =  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)
CmR
771 .. 1430  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
771 .. 1430  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
2862 .. 3450  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2862 .. 3450  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
7149 .. 7604  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
7149 .. 7604  =  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
ccdB
1772 .. 2077  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
ccdB
1772 .. 2077  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
CYC1 terminator
2367 .. 2614  =  248 bp
transcription terminator for CYC1
CYC1 terminator
2367 .. 2614  =  248 bp
transcription terminator for CYC1
URA3 promoter
5381 .. 5596  =  216 bp
URA3 promoter
5381 .. 5596  =  216 bp
attR1
511 .. 635  =  125 bp
recombination site for the Gateway® LR reaction
attR1
511 .. 635  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2118 .. 2242  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2118 .. 2242  =  125 bp
recombination site for the Gateway® LR reaction
V5 tag
2265 .. 2306  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
2265 .. 2306  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
687 .. 717  =  31 bp
   Segment 1:  -35  
   687 .. 692  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
687 .. 717  =  31 bp
   Segment 2:  
   693 .. 710  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
687 .. 717  =  31 bp
   Segment 3:  -10  
   711 .. 717  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
687 .. 717  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
475 .. 493  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
475 .. 493  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
2316 .. 2333  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2316 .. 2333  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
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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