pYES2

High-copy episomal vector for galactose-inducible expression of proteins in Saccharomyces cerevisiae.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsYeast PlasmidspYES2
NaeI (5711) NgoMIV (5709) SwaI (5383) SnaBI (5079) HpaI (4434) BmgBI (4374) PfoI * (4332) NruI (4028) BlpI (3992) BspDI - ClaI (3848) BmtI (3843) NheI (3839) NdeI (3699) BsgI (3670) SbfI (3636) BfuAI - BspMI (3625) XcmI (3487) NcoI (3405) BsmI (3254) ApaI * (3240) PspOMI * (3236) BsmBI - Esp3I (2988) AgeI (71) PvuII (471) T7 promoter HindIII (501) Acc65I (507) KpnI (511) Eco53kI (515) SacI (517) BamHI (519) EcoRI (550) BsaBI (564) NotI (577) PaeR7I - PspXI - XhoI (583) SphI (592) XbaI (595) BsrGI (774) MluI (781) BciVI (1239) BspHI (1756) AhdI (1929) BglI (2049) BsaHI (2466) pYES2 5856 bp
NaeI  (5711)
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  (5709)
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  (5383)
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  (5079)
1 site		 T A C G T A A T G C A T
HpaI  (4434)
1 site		 G T T A A C C A A T T G
BmgBI  (4374)
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  (4332)
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  (4028)
1 site		 T C G C G A A G C G C T
BlpI  (3992)
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  (3848)
1 site		 A T C G A T T A G C T A
ClaI  (3848)
1 site		 A T C G A T T A G C T A
BmtI  (3843)
1 site		 G C T A G C C G A T C G
NheI  (3839)
1 site		 G C T A G C C G A T C G
NdeI  (3699)
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  (3670)
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  (3636)
1 site		 C C T G C A G G G G A C G T C C
BfuAI  (3625)
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  (3625)
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  (3487)
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.
NcoI  (3405)
1 site		 C C A T G G G G T A C C
BsmI  (3254)
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  (3240)
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  (3236)
1 site		 G G G C C C C C C G G G
* Blocked by Dcm methylation.
BsmBI  (2988)
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2988)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
AgeI  (71)
1 site		 A C C G G T T G G C C A
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
Acc65I  (507)
1 site		 G G T A C C C C A T G G
KpnI  (511)
1 site		 G G T A C C C C A T G G
Eco53kI  (515)
1 site		 G A G C T C C T C G A G
SacI  (517)
1 site		 G A G C T C C T C G A G
BamHI  (519)
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.
EcoRI  (550)
1 site		 G A A T T C C T T A A G
BsaBI  (564)
1 site		 G A T N N N N A T C C T A N N N N T A G
NotI  (577)
1 site		 G C G G C C G C C G C C G G C G
PaeR7I  (583)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (583)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (583)
1 site		 C T C G A G G A G C T C
SphI  (592)
1 site		 G C A T G C C G T A C G
XbaI  (595)
1 site		 T C T A G A A G A T C T
BsrGI  (774)
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  (781)
1 site		 A C G C G T T G C G C A
BciVI  (1239)
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  (1756)
1 site		 T C A T G A A G T A C T
AhdI  (1929)
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  (2049)
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  (2466)
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.
2μ ori
4435 .. 5315  =  881 bp
yeast 2μ plasmid origin of replication
2μ ori
4435 .. 5315  =  881 bp
yeast 2μ plasmid origin of replication
AmpR
1856 .. 2716  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
   Segment 2:  
   1856 .. 2647  =  792 bp
   263 amino acids  =  29.1 kDa
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and related antibiotics
AmpR
1856 .. 2716  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
   Segment 1:  signal sequence  
   2648 .. 2716  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and related antibiotics
AmpR
1856 .. 2716  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and related antibiotics
URA3
2812 .. 3615  =  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
2812 .. 3615  =  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
1097 .. 1685  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1097 .. 1685  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5384 .. 5839  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5384 .. 5839  =  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
609 .. 856  =  248 bp
transcription terminator for CYC1
CYC1 terminator
609 .. 856  =  248 bp
transcription terminator for CYC1
URA3 promoter
3616 .. 3831  =  216 bp
URA3 promoter
3616 .. 3831  =  216 bp
MCS
501 .. 600  =  100 bp
multiple cloning site
MCS
501 .. 600  =  100 bp
multiple cloning site
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:  3104 .. 3580  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
ORF:  1986 .. 2252  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4470 .. 4763  =  294 bp
ORF:  97 amino acids  =  11.6 kDa
ORF:  2812 .. 3615  =  804 bp
ORF:  267 amino acids  =  29.3 kDa
ORF:  4216 .. 4455  =  240 bp
ORF:  79 amino acids  =  9.8 kDa
ORF:  1856 .. 2716  =  861 bp
ORF:  286 amino acids  =  31.7 kDa
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Individual Sequences & Maps

bleMX6
pFA6a-kanMX6
pPIC6(alpha) B
pYES2.1-E
hphMX6
pFA6a-kanMX6-PGAL1
pPIC6(alpha) C
pYES2 CT
kanMX
pFA6a-kanMX6-PGAL1-3HA
pPIC9
pYES2 NT A
kanMX2
pFA6a-kanMX6-PGAL1-GFP
pPIC9K
pYES2 NT B
kanMX4
pFA6a-kanMX6-PGAL1-GST
pPICZ A
pYES2 NT C
kanMX6
pFA6a-natMX6
pPICZ B
pYES3 CT
natMX6
pFA6a-TRP1
pPICZ C
pYES6 CT
pAbAi
pFA6a-TRP1-PGAL1
pPICZ(alpha) A
pYM25
pAD-GAL4
pFA6a-TRP1-PGAL1-3HA
pPICZ(alpha) B
pYM26
pAG25
pFA6a-TRP1-PGAL1-GFP
pPICZ(alpha) C
pYM44
pAG29
pFA6a-TRP1-PGAL1-GST
pPink-HC
YCp50
pAG32
pFLD
pPink-LC
YCp50-poly
pAO815
pFLD(alpha)
pPINK(alpha)-HC
YCplac111
patMX4
pGAD-C1
pRS303
YCplac22
pAUR101
pGAD-C2
pRS304
YCplac33
pAUR112
pGAD-C3
pRS305
YEp13
pAUR123
pGADT7 AD
pRS306
YEp24
pAUR224
pGADT7-Rec
pRS313
YEp351
pBridge
pGADT7-Rec (linearized)
pRS314
YEp352
pDH25
pGAPZ A
pRS315
YEp353
pEG202
pGAPZ B
pRS316
YEp354
pESC-HIS
pGAPZ C
pRS403
YEp355
pESC-LEU
pGAPZ(alpha) A
pRS404
YEp356
pESC-TRP
pGAPZ(alpha) B
pRS405
YEp356R
pESC-URA
pGAPZ(alpha) C
pRS406
YEp357
pFA6-kanMX4
pGBD-C1
pRS413
YEp357R
pFA6a
pGBD-C2
pRS414
YEp358
pFA6a-12Pk-kanMX6
pGBD-C3
pRS415
YEp358R
pFA6a-13Myc-His3MX6
pGBDU-C1
pRS416
YEp363
pFA6a-13Myc-kanMX6
pGBDU-C2
pRS423
YEp364
pFA6a-13Myc-TRP1
pGBDU-C3
pRS424
YEp365
pFA6a-3HA-His3MX6
pGBKT7
pRS425
YEp366
pFA6a-3HA-kanMX6
pGilda
pRS426
YEp366R
pFA6a-3HA-TRP1
pHIL-D2
pSpot5
YEp367
pFA6a-5FLAG-kanMX6
pHIL-S1
pSpot6
YEp367R
pFA6a-bleMX6
pJG4-5
pSpot7
YEp368
pFA6a-GFP(S65T)-His3MX6
pKLAC1
pSpot8
YEp368R
pFA6a-GFP(S65T)-kanMX6
pKLAC2
pTEF1 Zeo
YEplac112
pFA6a-GFP(S65T)-TRP1
pOAD
pUCATPH
YEplac181
pFA6a-GST-His3MX6
pOBD2
pY-secSUMOstar
YEplac195
pFA6a-GST-kanMX6
pP-secSUMO3
pY-SUMOstar
YIp351
pFA6a-GST-TRP1
pPIC3
pYC2-E
YIp352
pFA6a-His3MX6
pPIC3.5
pYC2 CT
YIp5
pFA6a-His3MX6-PGAL1
pPIC3.5K
pYC2 NT A
YIplac128
pFA6a-His3MX6-PGAL1-3HA
pPIC6 A
pYC2 NT B
YIplac204
pFA6a-His3MX6-PGAL1-GFP
pPIC6 B
pYC2 NT C
YIplac211
pFA6a-His3MX6-PGAL1-GST
pPIC6 C
pYC6 CT
YRp7
pFA6a-hphMX6
pPIC6(alpha) A
pYES2
alpha-factor secretion signal

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