YCplac33

Yeast centromeric plasmid with a URA3 marker. See also YCplac22 and YCplac111.
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HomePlasmidsYeast PlasmidsYCplac33
DrdI (5375) AhdI (4589) AatII (3670) ZraI (3668) NdeI (3449) XcmI (3237) PpuMI (3208) EcoRV (3176) NcoI (3155) BstBI (3097) BsmI (3004) ApaI * (2990) PspOMI * (2986) StuI (2928) lac operator HindIII (233) SphI (243) PstI - SbfI (249) SalI (251) XbaI (257) BamHI (263) TspMI - XmaI (268) SmaI (270) Acc65I (272) KpnI (276) Eco53kI (280) SacI (282) EcoRI (284) KasI (445) NarI (446) SfoI (447) PluTI (449) SpeI (550) PflFI - Tth111I (942) BsaBI (1516) NgoMIV (1771) NaeI (1773) NheI (1801) BmtI (1805) BtgZI (1913) BglII (1986) PaqCI (2303) BstXI (2446) BspDI * - ClaI * (2455) NsiI (2541) Bpu10I (2631) BsmBI - Esp3I (2738) YCplac33 5603 bp
DrdI  (5375)
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.
AhdI  (4589)
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.
AatII  (3670)
1 site		 G A C G T C C T G C A G
ZraI  (3668)
1 site		 G A C G T C C T G C A G
NdeI  (3449)
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.
XcmI  (3237)
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.
PpuMI  (3208)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
EcoRV  (3176)
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.
NcoI  (3155)
1 site		 C C A T G G G G T A C C
BstBI  (3097)
1 site		 T T C G A A A A G C T T
BsmI  (3004)
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.
ApaI  (2990)
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.
PspOMI  (2986)
1 site		 G G G C C C C C C G G G
* Blocked by Dcm methylation.
StuI  (2928)
1 site		 A G G C C T T C C G G A
HindIII  (233)
1 site		 A A G C T T T T C G A A
SphI  (243)
1 site		 G C A T G C C G T A C G
PstI  (249)
1 site		 C T G C A G G A C G T C
SbfI  (249)
1 site		 C C T G C A G G G G A C G T C C
SalI  (251)
1 site		 G T C G A C C A G C T G
XbaI  (257)
1 site		 T C T A G A A G A T C T
BamHI  (263)
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.
TspMI  (268)
1 site		 C C C G G G G G G C C C
XmaI  (268)
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  (270)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
Acc65I  (272)
1 site		 G G T A C C C C A T G G
KpnI  (276)
1 site		 G G T A C C C C A T G G
Eco53kI  (280)
1 site		 G A G C T C C T C G A G
SacI  (282)
1 site		 G A G C T C C T C G A G
EcoRI  (284)
1 site		 G A A T T C C T T A A G
KasI  (445)
1 site		 G G C G C C C C G C G G
NarI  (446)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (447)
1 site		 G G C G C C C C G C G G
PluTI  (449)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SpeI  (550)
1 site		 A C T A G T T G A T C A
PflFI  (942)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (942)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsaBI  (1516)
1 site		 G A T N N N N A T C C T A N N N N T A G
NgoMIV  (1771)
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.
NaeI  (1773)
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.
NheI  (1801)
1 site		 G C T A G C C G A T C G
BmtI  (1805)
1 site		 G C T A G C C G A T C G
BtgZI  (1913)
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.
BglII  (1986)
1 site		 A G A T C T T C T A G A
PaqCI  (2303)
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.
BstXI  (2446)
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.
BspDI  (2455)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2455)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
NsiI  (2541)
1 site		 A T G C A T T A C G T A
Bpu10I  (2631)
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.
BsmBI  (2738)
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  (2738)
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.
CEN/ARS
1064 .. 2227  =  1164 bp
S. cerevisiae CEN4 centromere fused to the autonomously replicating sequence ARS1/ARS416
CEN/ARS
1064 .. 2227  =  1164 bp
S. cerevisiae CEN4 centromere fused to the autonomously replicating sequence ARS1/ARS416
AmpR
3802 .. 4662  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3802 .. 3870  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3802 .. 4662  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3871 .. 4662  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3802 .. 4662  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
URA3
2562 .. 3365  =  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
2562 .. 3365  =  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
4833 .. 5421  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
4833 .. 5421  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
lacZα
216 .. 554  =  339 bp
112 amino acids  =  13.1 kDa
Product: LacZα fragment of β-galactosidase
lacZα
216 .. 554  =  339 bp
112 amino acids  =  13.1 kDa
Product: LacZα fragment of β-galactosidase
URA3 promoter
3366 .. 3586  =  221 bp
URA3 promoter
3366 .. 3586  =  221 bp
AmpR promoter
3697 .. 3801  =  105 bp
AmpR promoter
3697 .. 3801  =  105 bp
lac promoter
142 .. 172  =  31 bp
3 segments
   Segment 1:  -35  
   142 .. 147  =  6 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
   Segment 2:  
   148 .. 165  =  18 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
   Segment 3:  -10  
   166 .. 172  =  7 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
180 .. 196  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
180 .. 196  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
MCS
233 .. 289  =  57 bp
pUC19 multiple cloning site
MCS
233 .. 289  =  57 bp
pUC19 multiple cloning site
M13 rev
204 .. 220  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
204 .. 220  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
290 .. 306  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
290 .. 306  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  2854 .. 3330  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
ORF:  3802 .. 4662  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  216 .. 554  =  339 bp
ORF:  112 amino acids  =  13.1 kDa
ORF:  1980 .. 2330  =  351 bp
ORF:  116 amino acids  =  12.8 kDa
ORF:  2562 .. 3365  =  804 bp
ORF:  267 amino acids  =  29.3 kDa
ORF:  4266 .. 4532  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  143 .. 520  =  378 bp
ORF:  125 amino acids  =  14.4 kDa
ORF:  2066 .. 2476  =  411 bp
ORF:  136 amino acids  =  14.8 kDa
ORF:  2209 .. 2442  =  234 bp
ORF:  77 amino acids  =  8.9 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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