YEp363

Yeast episomal vector with a LEU2 marker for construction of lacZ fusions. For other reading frames, use YEp364 or YEp365.
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HomePlasmidsYeast PlasmidsYEp363
SmaI (8) TspMI - XmaI (6) BspQI - SapI (8346) AhdI (7341) BsaI (7275) ScaI (6860) BsaAI - SnaBI (5826) NsiI (5474) PpuMI (4885) BstEII (4740) KpnI (4532) Acc65I (4528) AgeI (4525) BamHI (11) SalI (17) PstI (27) HindIII (31) Bsu36I (259) Bpu10I (540) DraIII (1225) BsaBI * (1366) BclI * (1381) BssHII - MauBI (1533) Eco53kI (1973) SacI (1975) BstZ17I (2800) BsiWI (2806) NdeI (2992) BlpI (3048) BseRI (3435) BsrGI (3655) AflII (4460) YEp363 8406 bp
SmaI  (8)
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  (6)
1 site		 C C C G G G G G G C C C
XmaI  (6)
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.
BspQI  (8346)
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  (8346)
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.
AhdI  (7341)
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.
BsaI  (7275)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
ScaI  (6860)
1 site		 A G T A C T T C A T G A
BsaAI  (5826)
1 site		 Y A C G T R R T G C A Y
SnaBI  (5826)
1 site		 T A C G T A A T G C A T
NsiI  (5474)
1 site		 A T G C A T T A C G T A
PpuMI  (4885)
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.
BstEII  (4740)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
KpnI  (4532)
1 site		 G G T A C C C C A T G G
Acc65I  (4528)
1 site		 G G T A C C C C A T G G
AgeI  (4525)
1 site		 A C C G G T T G G C C A
BamHI  (11)
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.
SalI  (17)
1 site		 G T C G A C C A G C T G
PstI  (27)
1 site		 C T G C A G G A C G T C
HindIII  (31)
1 site		 A A G C T T T T C G A A
Bsu36I  (259)
1 site		 C C T N A G G G G A N T C C
Sticky ends from different Bsu36I sites may not be compatible.
Bpu10I  (540)
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.
DraIII  (1225)
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.
BsaBI  (1366)
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.
BclI  (1381)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BssHII  (1533)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
MauBI  (1533)
1 site		 C G C G C G C G G C G C G C G C
Eco53kI  (1973)
1 site		 G A G C T C C T C G A G
SacI  (1975)
1 site		 G A G C T C C T C G A G
BstZ17I  (2800)
1 site		 G T A T A C C A T A T G
BsiWI  (2806)
1 site		 C G T A C G G C A T G C
BsiWI is typically used at 55°C, but is 50% active at 37°C.
NdeI  (2992)
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.
BlpI  (3048)
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.
BseRI  (3435)
1 site		 G A G G A G ( N ) 8 N N C T C C T C ( N ) 8
Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BsrGI  (3655)
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.
AflII  (4460)
1 site		 C T T A A G G A A T T C
lacZ
48 .. 3095  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
lacZ
48 .. 3095  =  3048 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
LEU2
3685 .. 4779  =  1095 bp
364 amino acids  =  39.2 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
LEU2
3685 .. 4779  =  1095 bp
364 amino acids  =  39.2 kDa
Product: 3-isopropylmalate dehydrogenase, required for leucine biosynthesis
yeast auxotrophic marker
2μ ori
5185 .. 6167  =  983 bp
yeast 2μ plasmid origin of replication
2μ ori
5185 .. 6167  =  983 bp
yeast 2μ plasmid origin of replication
AmpR
6554 .. 7414  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6554 .. 6622  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6554 .. 7414  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6623 .. 7414  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6554 .. 7414  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
7585 .. 8173  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
7585 .. 8173  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
LEU2 promoter
4780 .. 5184  =  405 bp
LEU2 promoter
4780 .. 5184  =  405 bp
AmpR promoter
6449 .. 6553  =  105 bp
AmpR promoter
6449 .. 6553  =  105 bp
MCS
1 .. 36  =  36 bp
multiple cloning site
MCS
1 .. 36  =  36 bp
multiple cloning site
ORF:  2969 .. 3229  =  261 bp
ORF:  86 amino acids  =  10.1 kDa
ORF:  6554 .. 7414  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  297 .. 3095  =  2799 bp
ORF:  932 amino acids  =  105.9 kDa
ORF:  5220 .. 5510  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
ORF:  3685 .. 4779  =  1095 bp
ORF:  364 amino acids  =  39.2 kDa
ORF:  7018 .. 7284  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  894 .. 1208  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2319 .. 2555  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  1958 .. 2269  =  312 bp
ORF:  103 amino acids  =  10.4 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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