pYES2 NT A

High-copy episomal vector for galactose-inducible expression of N-terminally tagged proteins in S. cerevisiae. For other reading frames, use pYES2/NT B or pYES2/NT C.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsYeast PlasmidspYES2 NT A
SpeI (6033) NaeI (5893) NgoMIV (5891) SwaI (5565) SnaBI (5261) HpaI (4616) BmgBI (4556) PfoI * (4514) NruI (4210) BlpI (4174) BspDI - ClaI (4030) NdeI (3881) BsgI (3852) SbfI (3818) BfuAI - BspMI (3807) XcmI (3669) BsmI (3436) BsmBI - Esp3I (3170) GAL1 promoter PvuII (471) T7 promoter HindIII (501) ATG 6xHis Xpress™ tag Acc65I (602) AvrII - KpnI (606) BamHI (610) EcoRI (625) EagI - NotI (652) PaeR7I - PspXI - XhoI (658) XbaI (664) 6xHis PmeI (757) BsrGI (949) BciVI (1421) BspHI (1938) AhdI (2111) BglI (2231) BsaHI (2648) pYES2/NT A 6038 bp
SpeI  (6033)
1 site		 A C T A G T T G A T C A
NaeI  (5893)
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  (5891)
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  (5565)
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  (5261)
1 site		 T A C G T A A T G C A T
HpaI  (4616)
1 site		 G T T A A C C A A T T G
BmgBI  (4556)
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  (4514)
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  (4210)
1 site		 T C G C G A A G C G C T
BlpI  (4174)
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  (4030)
1 site		 A T C G A T T A G C T A
ClaI  (4030)
1 site		 A T C G A T T A G C T A
NdeI  (3881)
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  (3852)
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  (3818)
1 site		 C C T G C A G G G G A C G T C C
BfuAI  (3807)
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  (3807)
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  (3669)
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.
BsmI  (3436)
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.
BsmBI  (3170)
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  (3170)
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.
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  (602)
1 site		 G G T A C C C C A T G G
AvrII  (606)
1 site		 C C T A G G G G A T C C
KpnI  (606)
1 site		 G G T A C C C C A T G G
BamHI  (610)
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  (625)
1 site		 G A A T T C C T T A A G
EagI  (652)
1 site		 C G G C C G G C C G G C
NotI  (652)
1 site		 G C G G C C G C C G C C G G C G
PaeR7I  (658)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (658)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (658)
1 site		 C T C G A G G A G C T C
XbaI  (664)
1 site		 T C T A G A A G A T C T
PmeI  (757)
1 site		 G T T T A A A C C A A A T T T G
BsrGI  (949)
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.
BciVI  (1421)
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  (1938)
1 site		 T C A T G A A G T A C T
AhdI  (2111)
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  (2231)
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  (2648)
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
4617 .. 5497  =  881 bp
yeast 2μ plasmid origin of replication
2μ ori
4617 .. 5497  =  881 bp
yeast 2μ plasmid origin of replication
AmpR
2038 .. 2898  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
   Segment 2:  
   2038 .. 2829  =  792 bp
   263 amino acids  =  29.1 kDa
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and related antibiotics
AmpR
2038 .. 2898  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
   Segment 1:  signal sequence  
   2830 .. 2898  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and related antibiotics
AmpR
2038 .. 2898  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and related antibiotics
URA3
2994 .. 3797  =  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
2994 .. 3797  =  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
1279 .. 1867  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1279 .. 1867  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5566 .. 6021  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5566 .. 6021  =  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
784 .. 1031  =  248 bp
transcription terminator for CYC1
CYC1 terminator
784 .. 1031  =  248 bp
transcription terminator for CYC1
URA3 promoter
3798 .. 4013  =  216 bp
URA3 promoter
3798 .. 4013  =  216 bp
ATG
510 .. 512  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
510 .. 512  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
522 .. 539  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
522 .. 539  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
T7 tag (gene 10 leader)
543 .. 575  =  33 bp
11 amino acids  =  1.1 kDa
Product: leader peptide from bacteriophage T7 gene 10
promotes efficient translation in E. coli
T7 tag (gene 10 leader)
543 .. 575  =  33 bp
11 amino acids  =  1.1 kDa
Product: leader peptide from bacteriophage T7 gene 10
promotes efficient translation in E. coli
Xpress™ tag
579 .. 602  =  24 bp
8 amino acids  =  998.0 Da
2 segments
   Segment 1:  
   579 .. 587  =  9 bp
   3 amino acids  =  409.4 Da
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
Xpress™ tag
579 .. 602  =  24 bp
8 amino acids  =  998.0 Da
2 segments
   Segment 2:  enterokinase site  
   588 .. 602  =  15 bp
   5 amino acids  =  606.5 Da
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
Xpress™ tag
579 .. 602  =  24 bp
8 amino acids  =  998.0 Da
2 segments
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
V5 tag
682 .. 723  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
682 .. 723  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
733 .. 750  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
733 .. 750  =  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
MCS
602 .. 669  =  68 bp
multiple cloning site
MCS
602 .. 669  =  68 bp
multiple cloning site
ORF:  3286 .. 3762  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
ORF:  2168 .. 2434  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4652 .. 4945  =  294 bp
ORF:  97 amino acids  =  11.6 kDa
ORF:  900 .. 1178  =  279 bp
ORF:  92 amino acids  =  10.6 kDa
ORF:  2994 .. 3797  =  804 bp
ORF:  267 amino acids  =  29.3 kDa
ORF:  4398 .. 4637  =  240 bp
ORF:  79 amino acids  =  9.8 kDa
ORF:  2038 .. 2898  =  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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