pYES3 CT

High-copy episomal vector with a TRP1 marker for galactose-inducible expression of C-terminally tagged proteins in S. cerevisiae.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsYeast PlasmidspYES3 CT
NaeI (5725) NgoMIV (5723) DraIII (5622) PsiI (5494) SwaI (5397) SnaBI (5093) NsiI (4741) HpaI (4448) BmgBI (4388) PfoI * (4346) NruI (4042) BlpI (4006) StuI (3972) Bpu10I (3948) BtgI (3895) BspDI - ClaI (3862) BfuAI - BspMI - PaqCI (3565) Bsu36I (3374) MfeI (3272) BstZ17I (3099) PmlI (3092) TRP1 promoter AatII (2958) PvuII (471) T7 promoter HindIII (501) Acc65I (507) KpnI (511) Eco53kI (515) SacI (517) BamHI (519) EcoRI (550) NotI (577) PaeR7I - PspXI - XhoI (583) PspOMI (595) ApaI (599) PmeI (682) BsrGI (874) BspQI - SapI (1027) PciI (1143) AlwNI (1559) AhdI (2036) BglI (2156) ScaI (2516) ZraI (2956) pYES3/CT 5870 bp
NaeI  (5725)
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  (5723)
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.
DraIII  (5622)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
PsiI  (5494)
1 site		 T T A T A A A A T A T T
SwaI  (5397)
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  (5093)
1 site		 T A C G T A A T G C A T
NsiI  (4741)
1 site		 A T G C A T T A C G T A
HpaI  (4448)
1 site		 G T T A A C C A A T T G
BmgBI  (4388)
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  (4346)
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  (4042)
1 site		 T C G C G A A G C G C T
BlpI  (4006)
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.
StuI  (3972)
1 site		 A G G C C T T C C G G A
Bpu10I  (3948)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BtgI  (3895)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
BspDI  (3862)
1 site		 A T C G A T T A G C T A
ClaI  (3862)
1 site		 A T C G A T T A G C T A
BfuAI  (3565)
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  (3565)
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.
PaqCI  (3565)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
Bsu36I  (3374)
1 site		 C C T N A G G G G A N T C C
Sticky ends from different Bsu36I sites may not be compatible.
MfeI  (3272)
1 site		 C A A T T G G T T A A C
BstZ17I  (3099)
1 site		 G T A T A C C A T A T G
PmlI  (3092)
1 site		 C A C G T G G T G C A C
AatII  (2958)
1 site		 G A C G T C C T G C A G
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
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
PspOMI  (595)
1 site		 G G G C C C C C C G G G
ApaI  (599)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PmeI  (682)
1 site		 G T T T A A A C C A A A T T T G
BsrGI  (874)
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.
BspQI  (1027)
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  (1027)
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.
PciI  (1143)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AlwNI  (1559)
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  (2036)
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  (2156)
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.
ScaI  (2516)
1 site		 A G T A C T T C A T G A
ZraI  (2956)
1 site		 G A C G T C C T G C A G
2μ ori
4449 .. 5329  =  881 bp
yeast 2μ plasmid origin of replication
2μ ori
4449 .. 5329  =  881 bp
yeast 2μ plasmid origin of replication
AmpR
1963 .. 2823  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1963 .. 2754  =  792 bp
   263 amino acids  =  29.0 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1963 .. 2823  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2755 .. 2823  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1963 .. 2823  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
TRP1
3133 .. 3807  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
TRP1
3133 .. 3807  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
ori
1204 .. 1792  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1204 .. 1792  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5398 .. 5853  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5398 .. 5853  =  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
709 .. 956  =  248 bp
transcription terminator for CYC1
CYC1 terminator
709 .. 956  =  248 bp
transcription terminator for CYC1
AmpR promoter
2824 .. 2928  =  105 bp
AmpR promoter
2824 .. 2928  =  105 bp
TRP1 promoter
3031 .. 3132  =  102 bp
TRP1 promoter
3031 .. 3132  =  102 bp
MCS
501 .. 600  =  100 bp
multiple cloning site
MCS
501 .. 600  =  100 bp
multiple cloning site
V5 tag
607 .. 648  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
607 .. 648  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
658 .. 675  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
658 .. 675  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
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:  3133 .. 3807  =  675 bp
ORF:  224 amino acids  =  24.1 kDa
ORF:  2093 .. 2359  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3383 .. 3664  =  282 bp
ORF:  93 amino acids  =  10.8 kDa
ORF:  4484 .. 4777  =  294 bp
ORF:  97 amino acids  =  11.6 kDa
ORF:  825 .. 1103  =  279 bp
ORF:  92 amino acids  =  10.6 kDa
ORF:  4230 .. 4469  =  240 bp
ORF:  79 amino acids  =  9.8 kDa
ORF:  1963 .. 2823  =  861 bp
ORF:  286 amino acids  =  31.6 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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