pYES-DEST52

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

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspYES-DEST52
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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
2 segments
   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
2 segments
   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
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
lac UV5 promoter
687 .. 717  =  31 bp
3 segments
   Segment 1:  -35  
   687 .. 692  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
687 .. 717  =  31 bp
3 segments
   Segment 2:  
   693 .. 710  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
687 .. 717  =  31 bp
3 segments
   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
UAS
2 .. 119  =  118 bp
upstream activating sequence mediating Gal4-dependent induction
UAS
2 .. 119  =  118 bp
upstream activating sequence mediating Gal4-dependent induction
ORF:  3751 .. 4017  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6235 .. 6528  =  294 bp
ORF:  97 amino acids  =  11.6 kDa
ORF:  1772 .. 2077  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  2483 .. 2761  =  279 bp
ORF:  92 amino acids  =  10.6 kDa
ORF:  771 .. 1430  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4869 .. 5345  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
ORF:  4577 .. 5380  =  804 bp
ORF:  267 amino acids  =  29.3 kDa
ORF:  5981 .. 6220  =  240 bp
ORF:  79 amino acids  =  9.8 kDa
ORF:  3621 .. 4481  =  861 bp
ORF:  286 amino acids  =  31.7 kDa
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Individual Sequences & Maps

pAd BLOCK-iT-DEST
pDEST R4-R3 Vector II
pENTR11
pHELLSGATE 4
pAd CMV V5-DEST
pDEST10
pENTR1A
pHELLSGATE 8
pAd PL-DEST
pDEST14
pENTR2B
pIB V5-His-DEST
pBAD-DEST49
pDEST15
pENTR3C
pLenti4 BLOCK-iT-DEST
pcDNA-DEST40
pDEST17
pENTR4
pLenti4 V5-DEST
pcDNA-DEST47
pDEST20
pET-53-DEST
pLenti6.2 C-Lumio V5-DEST
pcDNA-DEST53
pDEST24
pET-54-DEST
pLenti6.2 N-Lumio V5-DEST
pcDNA3.1 nV5-DEST
pDEST26
pET-55-DEST
pLenti6.2 V5-DEST
pcDNA3.2 capTEV-CT V5-DEST
pDEST27
pET-56-DEST
pLenti6.3 TO V5-DEST
pcDNA3.2 capTEV-NT V5-DEST
pDEST8
pET-57-DEST
pLenti6.3 V5-DEST
pcDNA3.2 V5-DEST
pDONR221 P5-P2
pET-58-DEST
pLenti6.4 R4R2 V5-DEST
pcDNA6.2 C-EmGFP-DEST
pDONR P2r-P3
pET-59-DEST
pLenti6 BLOCK-iT-DEST
pcDNA6.2 C-YFP-DEST
pDONR P4-P1r
pET-60-DEST
pLenti6 capTEV-CT-DEST
pcDNA6.2 cGeneBLAzer-DEST
pDONR Zeo
pET-61-DEST
pLenti6 capTEV-NT-DEST1
pcDNA6.2 cLumio-DEST
pDONR201
pET-62-DEST
pLenti6 UbC V5-DEST
pcDNA6.2 EmGFP-Bsd V5-DEST
pDONR207
pET-DEST42
pLenti6 V5-DEST
pcDNA6.2 N-EmGFP-DEST
pDONR221
pET104-DEST
pLenti7.3 V5-DEST
pcDNA6.2 N-YFP-DEST
pDONR221 P1-P4
pET104.1-DEST
pMT-DEST48
pcDNA6.2 nGeneBLAzer-DEST
pDONR221 P1-P5r
pET160-DEST
pMT BioEase-DEST
pcDNA6.2 nLumio-DEST
pDONR221 P3-P2
pET161-DEST
pSCREEN-iT lacZ-DEST
pcDNA6.2 V5-DEST
pDONR221 P4r-P3r
pEXP1-DEST
pT-REx-DEST30
pcDNA6.2 V5-pL-DEST
pDONR221 P5-P4
pEXP2-DEST
pT-REx-DEST31
pCMV SPORT6
pDONR222
pEXP3-DEST
pYES-DEST52
pCMV SPORT6.1
pDONR223
pEXP4-DEST
Reading Frame Cassette A
pCOLA-2-DEST
pEF-DEST51
pHELLSGATE
Reading Frame Cassette B
pCOLA-3-DEST
pEF5 FRT V5-DEST
pHELLSGATE 12
Reading Frame Cassette C.1
pDEST R4-R3
pENTR GeneBLAzer

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