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pYC6/CT

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

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pYC6 CT Sequence and MappYC6 CT.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 NaeI (4359) MreI - NgoMIV - SgrAI (4357) SwaI (3876) AanI - PsiI (3870) BsaAI - PmlI (3852) BspDI - ClaI (3832) BmtI (3827) NheI (3823) HincII (3404) BtgI - NcoI - StyI (3338) MscI (3337) NruI * (3109) BsaBI * (3042) BlpI (2942) DraIII (2938) BmgBI (2911) SmaI (2906) TspMI - XmaI (2904) BglII (2898) AccI (2889) BsaHI (2573) ScaI (2516) BstAPI (323) SspI (497) HindIII (501) Acc65I (507) KpnI (511) Eco53kI (515) SacI (517) BamHI (519) BstXI (546) EcoRI (550) PstI (559) EcoRV (562) BstXI (572) NotI (577) PaeR7I - PspXI - XhoI (583) XbaI (589) PspOMI (595) ApaI (599) BstBI (602) PmeI (682) BsrGI (874) PciI (1143) BciVI (1346) AlwNI (1559) BspHI (1863) AhdI (2036) BpmI (2106) BglI (2156) pYC6/CT 4504 bp
NaeI  (4359)
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  (4357)
1 site
C G C C G G C G G C G G C C G C
NgoMIV  (4357)
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  (4357)
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  (3876)
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  (3870)
1 site
T T A T A A A A T A T T
PsiI  (3870)
1 site
T T A T A A A A T A T T
BsaAI  (3852)
1 site
Y A C G T R R T G C A Y
PmlI  (3852)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
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
HincII  (3404)
1 site
G T Y R A C C A R Y T G
BtgI  (3338)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
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
NruI  (3109)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
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.
BlpI  (2942)
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.
DraIII  (2938)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BmgBI  (2911)
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.
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.
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.
BglII  (2898)
1 site
A G A T C T T C T A G A
AccI  (2889)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
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.
ScaI  (2516)
1 site
A G T A C T T C A T G A
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.
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  (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.
BstXI  (546)
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  (550)
1 site
G A A T T C C T T A A G
PstI  (559)
1 site
C T G C A G G A C G T C
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.
BstXI  (572)
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.
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.
BstBI  (602)
1 site
T T C G A A A A G C T T
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.
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.
BpmI  (2106)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
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.
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
CEN/ARS
3848 .. 4351  =  504 bp
S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence
CEN/ARS
3848 .. 4351  =  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
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
495 .. 606  =  112 bp
multiple cloning site
MCS
495 .. 606  =  112 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
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