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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.

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pYC2 NT C Sequence and MappYC2 NT C.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 SpeI (4696) NaeI (4556) MreI - NgoMIV - SgrAI (4554) SwaI (4073) AanI - AanI (4067) BsaAI - PmlI (4049) BspDI - ClaI (4029) NdeI (3880) BsgI (3851) SbfI (3817) BfuAI - BspMI (3806) XcmI (3668) BsmI (3435) StuI (3359) BsmBI (3169) Bpu10I (3062) AanI - AanI (2980) NsiI (2972) BsaHI (2647) PvuI (2480) GAL1 promoter BtgZI (297) BstAPI (323) PvuII (471) SspI (497) HindIII (501) ATG 6xHis Acc65I (602) KpnI (606) BamHI (609) BstXI (620) EcoRI (624) BstXI (646) EagI - NotI (651) AvaI - BsoBI - PaeR7I - PspXI - XhoI (657) BmeT110I (658) XbaI (663) V5 tag 6xHis PmeI (756) 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.
AanI  (4067)
2 sites
T T A T A A A A T A T T
AanI  (4067)
2 sites
T T A T A A A A T A T T
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

PmlI gradually loses activity when stored at -20°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.
Bpu10I  (3062)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
AanI  (2980)
2 sites
T T A T A A A A T A T T
AanI  (2980)
2 sites
T T A T A A A A T A T T
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.
BstXI  (620)
2 sites
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.
EcoRI  (624)
1 site
G A A T T C C T T A A G
BstXI  (646)
2 sites
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.
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
BmeT110I  (658)
1 site
C Y C G R G G R G C Y 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
   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
   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
URA3 promoter
3797 .. 4012  =  216 bp
URA3 promoter
3797 .. 4012  =  216 bp
MCS
602 .. 668  =  67 bp
multiple cloning site
MCS
602 .. 668  =  67 bp
multiple cloning 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
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
T7 promoter
475 .. 493  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
475 .. 493  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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
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
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
UAS
2 .. 119  =  118 bp
upstream activating sequence mediating
Gal4-dependent induction
UAS
2 .. 119  =  118 bp
upstream activating sequence mediating
Gal4-dependent induction
Xpress™ tag
579 .. 602  =  24 bp
8 amino acids  =  998.0 Da
   Segment 1:  
   579 .. 587  =  9 bp
   3 amino acids  =  409.5 Da
Product: Xpress™ epitope tag, including an
enterokinase recognition and cleavage site
Xpress™ tag
579 .. 602  =  24 bp
8 amino acids  =  998.0 Da
   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
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