YCp50

Yeast centromere-containing shuttle vector with ampicillin-resistance and tetracycline-resistance markers.
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AatII (7880) ZraI (7878) PvuI (7328) AseI (7130) AhdI (6958) BsaAI (5819) PfoI (5709) BglII (5417) PaeR7I - XhoI (4702) BstXI (4656) KpnI (4231) Acc65I (4227) MfeI (4164) EcoRI (1) tet promoter BspDI - ClaI (27) HindIII (32) BamHI (378) SgrAI (413) SphI (569) EcoNI (629) SalI (654) PshAI (719) EagI (942) NruI (977) NcoI (1866) BstBI (1926) PspOMI * (2035) ApaI * (2039) NsiI (2488) TspMI - XmaI (2540) SmaI (2542) MscI * (2627) BspEI * (2845) PaqCI (3237) SpeI (3294) HpaI (4101) YCp50 7950 bp
AatII  (7880)
1 site
G A C G T C C T G C A G
ZraI  (7878)
1 site
G A C G T C C T G C A G
PvuI  (7328)
1 site
C G A T C G G C T A G C
AseI  (7130)
1 site
A T T A A T T A A T T A
AhdI  (6958)
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.
BsaAI  (5819)
1 site
Y A C G T R R T G C A Y
PfoI  (5709)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BglII  (5417)
1 site
A G A T C T T C T A G A
PaeR7I  (4702)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (4702)
1 site
C T C G A G G A G C T C
BstXI  (4656)
1 site
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.
KpnI  (4231)
1 site
G G T A C C C C A T G G
Acc65I  (4227)
1 site
G G T A C C C C A T G G
MfeI  (4164)
1 site
C A A T T G G T T A A C
EcoRI  (1)
1 site
G A A T T C C T T A A G
BspDI  (27)
1 site
A T C G A T T A G C T A
ClaI  (27)
1 site
A T C G A T T A G C T A
HindIII  (32)
1 site
A A G C T T T T C G A A
BamHI  (378)
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.
SgrAI  (413)
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.
SphI  (569)
1 site
G C A T G C C G T A C G
EcoNI  (629)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
SalI  (654)
1 site
G T C G A C C A G C T G
PshAI  (719)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
EagI  (942)
1 site
C G G C C G G C C G G C
NruI  (977)
1 site
T C G C G A A G C G C T
NcoI  (1866)
1 site
C C A T G G G G T A C C
BstBI  (1926)
1 site
T T C G A A A A G C T T
PspOMI  (2035)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI  (2039)
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.
NsiI  (2488)
1 site
A T G C A T T A C G T A
TspMI  (2540)
1 site
C C C G G G G G G C C C
XmaI  (2540)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2542)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
MscI  (2627)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
BspEI  (2845)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
PaqCI  (3237)
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.
SpeI  (3294)
1 site
A C T A G T T G A T C A
HpaI  (4101)
1 site
G T T A A C C A A T T G
CEN/ARS
3809 .. 5658  =  1850 bp
S. cerevisiae CEN4 centromere fused to the autonomously replicating sequence ARS1/ARS416
CEN/ARS
3809 .. 5658  =  1850 bp
S. cerevisiae CEN4 centromere fused to the autonomously replicating sequence ARS1/ARS416
TcR
89 .. 1279  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
89 .. 1279  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
AmpR
6885 .. 7745  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   6885 .. 7676  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6885 .. 7745  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   7677 .. 7745  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6885 .. 7745  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
URA3
1661 .. 2464  =  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
1661 .. 2464  =  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
6126 .. 6714  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
6126 .. 6714  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
URA3 promoter
1445 .. 1660  =  216 bp
URA3 promoter
1445 .. 1660  =  216 bp
AmpR promoter
7746 .. 7850  =  105 bp
AmpR promoter
7746 .. 7850  =  105 bp
tet promoter
13 .. 41  =  29 bp
3 segments
   Segment 1:  -35  
   13 .. 18  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
13 .. 41  =  29 bp
3 segments
   Segment 2:  
   19 .. 35  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
13 .. 41  =  29 bp
3 segments
   Segment 3:  -10  
   36 .. 41  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
13 .. 41  =  29 bp
3 segments
E. coli promoter for tetracycline efflux protein gene
ORF:  262 .. 645  =  384 bp
ORF:  127 amino acids  =  14.7 kDa
ORF:  1183 .. 1506  =  324 bp
ORF:  107 amino acids  =  12.1 kDa
ORF:  7015 .. 7281  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  89 .. 1279  =  1191 bp
ORF:  396 amino acids  =  41.5 kDa
ORF:  1661 .. 2464  =  804 bp
ORF:  267 amino acids  =  29.3 kDa
ORF:  2999 .. 3298  =  300 bp
ORF:  99 amino acids  =  12.1 kDa
ORF:  5411 .. 5698  =  288 bp
ORF:  95 amino acids  =  10.7 kDa
ORF:  442 .. 792  =  351 bp
ORF:  116 amino acids  =  12.2 kDa
ORF:  1696 .. 2172  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
ORF:  6885 .. 7745  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  326 .. 568  =  243 bp
ORF:  80 amino acids  =  8.0 kDa
ORF:  626 .. 1084  =  459 bp
ORF:  152 amino acids  =  16.7 kDa
ORF:  2696 .. 3094  =  399 bp
ORF:  132 amino acids  =  15.3 kDa
ORF:  4283 .. 4939  =  657 bp
ORF:  218 amino acids  =  25.1 kDa
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Individual Sequences & Maps

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