pOBD2

Yeast two-hybrid "bait" vector for fusing a gene to the GAL4 DNA binding domain.

Sequence Author: Fields Lab

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PacI (6987) BsmI (6627) Bpu10I (6593) PfoI (6323) BspQI - SapI (6257) DrdI (6038) AlwNI (5731) AhdI (5252) AatII (4333) ZraI (4331) EcoO109I (4272) TRP1 promoter PmlI (4192) BstAPI (4076) Bsu36I (3907) PaqCI (3715) BanII (3648) PaeR7I - XhoI (753) HpaI (813) EcoRI (1028) PvuII (1036) NcoI (1044) TspMI - XmaI (1055) SmaI (1057) SalI (1066) PstI (1076) BseRI (1184) MscI (1214) SpeI (1962) PflFI - Tth111I (2354) DraIII (2840) NheI (3213) BmtI (3217) pOBD2 7269 bp
PacI  (6987)
1 site
T T A A T T A A A A T T A A T T
BsmI  (6627)
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.
Bpu10I  (6593)
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.
PfoI  (6323)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BspQI  (6257)
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  (6257)
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.
DrdI  (6038)
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.
AlwNI  (5731)
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  (5252)
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  (4333)
1 site
G A C G T C C T G C A G
ZraI  (4331)
1 site
G A C G T C C T G C A G
EcoO109I  (4272)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PmlI  (4192)
1 site
C A C G T G G T G C A C
BstAPI  (4076)
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.
Bsu36I  (3907)
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.
PaqCI  (3715)
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.
BanII  (3648)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PaeR7I  (753)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (753)
1 site
C T C G A G G A G C T C
HpaI  (813)
1 site
G T T A A C C A A T T G
EcoRI  (1028)
1 site
G A A T T C C T T A A G
PvuII  (1036)
1 site
C A G C T G G T C G A C
NcoI  (1044)
1 site
C C A T G G G G T A C C
TspMI  (1055)
1 site
C C C G G G G G G C C C
XmaI  (1055)
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  (1057)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SalI  (1066)
1 site
G T C G A C C A G C T G
PstI  (1076)
1 site
C T G C A G G A C G T C
BseRI  (1184)
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.
MscI  (1214)
1 site
T G G C C A A C C G G T
SpeI  (1962)
1 site
A C T A G T T G A T C A
PflFI  (2354)
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  (2354)
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.
DraIII  (2840)
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.
NheI  (3213)
1 site
G C T A G C C G A T C G
BmtI  (3217)
1 site
G C T A G C C G A T C G
CEN/ARS
2477 .. 3639  =  1163 bp
S. cerevisiae CEN4 centromere fused to the autonomously replicating sequence ARS1/ARS416
CEN/ARS
2477 .. 3639  =  1163 bp
S. cerevisiae CEN4 centromere fused to the autonomously replicating sequence ARS1/ARS416
AmpR
4465 .. 5325  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4465 .. 4533  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4465 .. 5325  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4534 .. 5325  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4465 .. 5325  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ADH1 promoter
7073 .. 508  =  705 bp
promoter for alcohol dehydrogenase 1
ADH1 promoter
7073 .. 508  =  705 bp
promoter for alcohol dehydrogenase 1
ori
5496 .. 6084  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
5496 .. 6084  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
GAL4 DNA binding domain
536 .. 976  =  441 bp
147 amino acids  =  16.9 kDa
Product: DNA binding domain of the GAL4 transcriptional activator
GAL4 DNA binding domain
536 .. 976  =  441 bp
147 amino acids  =  16.9 kDa
Product: DNA binding domain of the GAL4 transcriptional activator
ADH1 terminator
1458 .. 1645  =  188 bp
transcription terminator for alcohol dehydrogenase 1
ADH1 terminator
1458 .. 1645  =  188 bp
transcription terminator for alcohol dehydrogenase 1
AmpR promoter
4360 .. 4464  =  105 bp
AmpR promoter
4360 .. 4464  =  105 bp
TRP1 promoter
4153 .. 4254  =  102 bp
TRP1 promoter
4153 .. 4254  =  102 bp
TRP1
3478 .. 4152  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
TRP1
3478 .. 4152  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
ORF:  4465 .. 5325  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  6370 .. 6711  =  342 bp
ORF:  113 amino acids  =  12.8 kDa
ORF:  536 .. 1087  =  552 bp
ORF:  183 amino acids  =  21.0 kDa
ORF:  3392 .. 3742  =  351 bp
ORF:  116 amino acids  =  12.8 kDa
ORF:  3478 .. 4152  =  675 bp
ORF:  224 amino acids  =  24.1 kDa
ORF:  3621 .. 3902  =  282 bp
ORF:  93 amino acids  =  10.8 kDa
ORF:  4929 .. 5195  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Download pOBD2.dna file

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