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Plasmid Files

pYES6/CT

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

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pYES6 CT.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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NaeI (5695) NgoMIV (5693) PsiI (5464) SwaI (5367) SnaBI (5063) NsiI (4711) HpaI (4418) PfoI * (4316) StuI (3942) Bpu10I (3918) BspDI - ClaI (3832) BmtI (3827) NheI (3823) NcoI - StyI (3338) MscI (3337) BsaBI * (3042) SmaI (2906) HindIII (501) Acc65I (507) KpnI (511) Eco53kI (515) SacI (517) BamHI (519) EcoRI (550) EcoRV (562) NotI (577) PaeR7I - PspXI - XhoI (583) XbaI (589) PspOMI (595) ApaI (599) PmeI (682) BsrGI (874) BspQI - SapI (1027) PciI (1143) BciVI (1346) AlwNI (1559) BspHI (1863) AhdI (2036) BglI (2156) ScaI (2516) BsaHI (2573) BglII (2898) TspMI - XmaI (2904) pYES6/CT 5840 bp
NaeI  (5695)
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  (5693)
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.
PsiI  (5464)
1 site
T T A T A A A A T A T T
SwaI  (5367)
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  (5063)
1 site
T A C G T A A T G C A T
NsiI  (4711)
1 site
A T G C A T T A C G T A
HpaI  (4418)
1 site
G T T A A C C A A T T G
PfoI  (4316)
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.
StuI  (3942)
1 site
A G G C C T T C C G G A
Bpu10I  (3918)
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.
BspDI  (3832)
1 site
A T C G A T T A G C T A
ClaI  (3832)
1 site
A T C G A T T A G C T A
BmtI  (3827)
1 site
G C T A G C C G A T C G
NheI  (3823)
1 site
G C T A G C C G A T C G
NcoI  (3338)
1 site
C C A T G G G G T A C C
StyI  (3338)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
MscI  (3337)
1 site
T G G C C A A C C G G T
BsaBI  (3042)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
SmaI  (2906)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
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
EcoRV  (562)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
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
XbaI  (589)
1 site
T C T A G A A G A T C T
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.
BciVI  (1346)
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.
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.
BspHI  (1863)
1 site
T C A T G A A G T A C T
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
BsaHI  (2573)
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.
BglII  (2898)
1 site
A G A T C T T C T A G A
TspMI  (2904)
1 site
C C C G G G G G G C C C
XmaI  (2904)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
2μ ori
4419 .. 5299  =  881 bp
yeast 2μ plasmid origin of replication
2μ ori
4419 .. 5299  =  881 bp
yeast 2μ plasmid origin of replication
AmpR
1963 .. 2823  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1963 .. 2754  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1963 .. 2823  =  861 bp
286 amino acids  =  31.6 kDa
   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
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
5368 .. 5823  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5368 .. 5823  =  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
TEF1 promoter
3414 .. 3821  =  408 bp
promoter for EF-1α
TEF1 promoter
3414 .. 3821  =  408 bp
promoter for EF-1α
BSD
2943 .. 3341  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
2943 .. 3341  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CYC1 terminator
709 .. 956  =  248 bp
transcription terminator for CYC1
CYC1 terminator
709 .. 956  =  248 bp
transcription terminator for CYC1
MCS
501 .. 600  =  100 bp
multiple cloning site
MCS
501 .. 600  =  100 bp
multiple cloning site
EM7 promoter
3360 .. 3407  =  48 bp
synthetic bacterial promoter
EM7 promoter
3360 .. 3407  =  48 bp
synthetic bacterial promoter
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
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
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
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:  2093 .. 2359  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4454 .. 4747  =  294 bp
ORF:  97 amino acids  =  11.6 kDa
ORF:  825 .. 1103  =  279 bp
ORF:  92 amino acids  =  10.6 kDa
ORF:  3039 .. 3395  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  2943 .. 3341  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  4200 .. 4439  =  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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