pDH25

Plasmid for transforming fungi such as Aspergillus nidulans to hygromycin resistance.
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HomePlasmidsYeast PlasmidspDH25
XbaI (6987) PasI (6748) BseRI (6690) BlpI (6566) SacII (6015) RsrII (5643) AsiSI (5599) BspDI - ClaI (5230) AflII (5167) SalI (4869) SmaI (4579) TspMI - XmaI (4577) BclI * (4546) EcoRV (4479) BmgBI (4339) BstBI (4017) SspI (3795) SgrAI (35) SphI (191) EcoNI (251) SalI (276) NruI (599) MscI * (1071) FspAI (1081) Bpu10I (1206) PvuII (1691) BsaAI (1852) BspQI - SapI (1982) PciI (2098) AlwNI (2514) AhdI (2991) pDH25 6993 bp
XbaI  (6987)
1 site		 T C T A G A A G A T C T
PasI  (6748)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
BseRI  (6690)
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.
BlpI  (6566)
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.
SacII  (6015)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
RsrII  (5643)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
AsiSI  (5599)
1 site		 G C G A T C G C C G C T A G C G
BspDI  (5230)
1 site		 A T C G A T T A G C T A
ClaI  (5230)
1 site		 A T C G A T T A G C T A
AflII  (5167)
1 site		 C T T A A G G A A T T C
SalI  (4869)
2 sites		 G T C G A C C A G C T G
SmaI  (4579)
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  (4577)
1 site		 C C C G G G G G G C C C
XmaI  (4577)
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.
BclI  (4546)
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.
EcoRV  (4479)
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.
BmgBI  (4339)
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.
BstBI  (4017)
1 site		 T T C G A A A A G C T T
SspI  (3795)
1 site		 A A T A T T T T A T A A
SgrAI  (35)
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  (191)
1 site		 G C A T G C C G T A C G
EcoNI  (251)
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  (276)
2 sites		 G T C G A C C A G C T G
NruI  (599)
1 site		 T C G C G A A G C G C T
MscI  (1071)
1 site		 T G G C C A A C C G G T
* Blocked by Dcm methylation.
FspAI  (1081)
1 site		 R T G C G C A Y Y A C G C G T R
Bpu10I  (1206)
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.
PvuII  (1691)
1 site		 C A G C T G G T C G A C
BsaAI  (1852)
1 site		 Y A C G T R R T G C A Y
BspQI  (1982)
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  (1982)
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  (2098)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AlwNI  (2514)
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.
AhdI  (2991)
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.
HygR
5235 .. 6260  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
HygR
5235 .. 6260  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
AmpR
2918 .. 3778  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   2918 .. 3709  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2918 .. 3778  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   3710 .. 3778  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2918 .. 3778  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2159 .. 2747  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2159 .. 2747  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
trpC terminator
6429 .. 6987  =  559 bp
terminator for Aspergillus nidulans trpC
trpC terminator
6429 .. 6987  =  559 bp
terminator for Aspergillus nidulans trpC
trpC promoter
4870 .. 5230  =  361 bp
promoter for Aspergillus nidulans trpC
trpC promoter
4870 .. 5230  =  361 bp
promoter for Aspergillus nidulans trpC
rop
1540 .. 1731  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
1540 .. 1731  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
AmpR promoter
3779 .. 3883  =  105 bp
AmpR promoter
3779 .. 3883  =  105 bp
ORF:  805 .. 1143  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  1507 .. 1731  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  116 .. 901  =  786 bp
ORF:  261 amino acids  =  27.5 kDa
ORF:  3048 .. 3314  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5235 .. 6260  =  1026 bp
ORF:  341 amino acids  =  38.0 kDa
ORF:  64 .. 414  =  351 bp
ORF:  116 amino acids  =  12.2 kDa
ORF:  1140 .. 1508  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  5214 .. 5993  =  780 bp
ORF:  259 amino acids  =  29.2 kDa
ORF:  6920 .. 190  =  264 bp
ORF:  87 amino acids  =  9.0 kDa
ORF:  248 .. 706  =  459 bp
ORF:  152 amino acids  =  16.7 kDa
ORF:  2918 .. 3778  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  6059 .. 6301  =  243 bp
ORF:  80 amino acids  =  8.4 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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