pYC2 NT C

Centromeric vector for galactose-inducible expression of N-terminally tagged proteins in S. cerevisiae. For other reading frames, use pYC2/NT A or pYC2/NT B.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsYeast PlasmidspYC2 NT C
SpeI (4696) NaeI (4556) MreI - NgoMIV - SgrAI (4554) SwaI (4073) BsaAI - PmlI (4049) BspDI - ClaI (4029) NdeI (3880) BsgI (3851) SbfI (3817) BfuAI - BspMI (3806) XcmI (3668) BsmI (3435) StuI (3359) BsmBI - Esp3I (3169) Bpu10I (3062) NsiI (2972) BsaHI (2647) PvuI (2480) GAL1 promoter BtgZI (297) BstAPI (323) PvuII (471) T7 promoter SspI (497) HindIII (501) ATG 6xHis Xpress™ tag Acc65I (602) KpnI (606) BamHI (609) EcoRI (624) EagI - NotI (651) AvaI - BsoBI - PaeR7I - PspXI - XhoI (657) XbaI (663) 6xHis PmeI (756) CYC1 terminator BsrGI (948) DrdI (1325) BciVI (1420) BspHI (1937) AhdI (2110) AfeI (2140) BglI (2230) FspI (2332) pYC2/NT C 4701 bp
SpeI  (4696)
1 site		 A C T A G T T G A T C A
NaeI  (4556)
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.
MreI  (4554)
1 site		 C G C C G G C G G C G G C C G C
NgoMIV  (4554)
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.
SgrAI  (4554)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
SwaI  (4073)
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.
BsaAI  (4049)
1 site		 Y A C G T R R T G C A Y
PmlI  (4049)
1 site		 C A C G T G G T G C A C
BspDI  (4029)
1 site		 A T C G A T T A G C T A
ClaI  (4029)
1 site		 A T C G A T T A G C T A
NdeI  (3880)
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  (3851)
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  (3817)
1 site		 C C T G C A G G G G A C G T C C
BfuAI  (3806)
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  (3806)
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  (3668)
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  (3435)
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.
StuI  (3359)
1 site		 A G G C C T T C C G G A
BsmBI  (3169)
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  (3169)
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.
Bpu10I  (3062)
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.
NsiI  (2972)
1 site		 A T G C A T T A C G T A
BsaHI  (2647)
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.
PvuI  (2480)
1 site		 C G A T C G G C T A G C
BtgZI  (297)
1 site		 G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4
Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BstAPI  (323)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
PvuII  (471)
1 site		 C A G C T G G T C G A C
SspI  (497)
1 site		 A A T A T T T T A T A A
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
KpnI  (606)
1 site		 G G T A C C C C A T G G
BamHI  (609)
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  (624)
1 site		 G A A T T C C T T A A G
EagI  (651)
1 site		 C G G C C G G C C G G C
NotI  (651)
1 site		 G C G G C C G C C G C C G G C G
AvaI  (657)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (657)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (657)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (657)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (657)
1 site		 C T C G A G G A G C T C
XbaI  (663)
1 site		 T C T A G A A G A T C T
PmeI  (756)
1 site		 G T T T A A A C C A A A T T T G
BsrGI  (948)
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.
DrdI  (1325)
1 site		 G A C N N N N N N G T C C T G N N N N N N C A G
Sticky ends from different DrdI sites may not be compatible.
BciVI  (1420)
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  (1937)
1 site		 T C A T G A A G T A C T
AhdI  (2110)
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.
AfeI  (2140)
1 site		 A G C G C T T C G C G A
BglI  (2230)
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.
FspI  (2332)
1 site		 T G C G C A A C G C G T
AmpR
2037 .. 2897  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
   Segment 2:  
   2037 .. 2828  =  792 bp
   263 amino acids  =  29.1 kDa
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and related antibiotics
AmpR
2037 .. 2897  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
   Segment 1:  signal sequence  
   2829 .. 2897  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and related antibiotics
AmpR
2037 .. 2897  =  861 bp
286 amino acids  =  31.7 kDa
2 segments
Product: β-lactamase
confers resistance fo ampicillin, carbenicillin, and related antibiotics
URA3
2993 .. 3796  =  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
2993 .. 3796  =  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
1278 .. 1866  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1278 .. 1866  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CEN/ARS
4045 .. 4548  =  504 bp
S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence
CEN/ARS
4045 .. 4548  =  504 bp
S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence
GAL1 promoter
2 .. 443  =  442 bp
inducible promoter, regulated by Gal4
GAL1 promoter
2 .. 443  =  442 bp
inducible promoter, regulated by Gal4
CYC1 terminator
783 .. 1030  =  248 bp
transcription terminator for CYC1
CYC1 terminator
783 .. 1030  =  248 bp
transcription terminator for CYC1
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
681 .. 722  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
681 .. 722  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
732 .. 749  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
732 .. 749  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
URA3 promoter
3797 .. 4012  =  216 bp
URA3 promoter
3797 .. 4012  =  216 bp
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 .. 668  =  67 bp
multiple cloning site
MCS
602 .. 668  =  67 bp
multiple cloning site
ORF:  592 .. 858  =  267 bp
ORF:  88 amino acids  =  9.7 kDa
ORF:  2167 .. 2433  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  899 .. 1177  =  279 bp
ORF:  92 amino acids  =  10.6 kDa
ORF:  510 .. 752  =  243 bp
ORF:  80 amino acids  =  9.0 kDa
ORF:  3285 .. 3761  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
ORF:  465 .. 749  =  285 bp
ORF:  94 amino acids  =  10.9 kDa
ORF:  2037 .. 2897  =  861 bp
ORF:  286 amino acids  =  31.7 kDa
ORF:  2993 .. 3796  =  804 bp
ORF:  267 amino acids  =  29.3 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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