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Plasmid Files

pPTR II

Autonomously replicating E. coli-Aspergillus shuttle vector with the ptrA gene conferring resistance to pyrithiamine.            

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pPTR II.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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NaeI (9900) NgoMIV (9898) KpnI (7045) Acc65I (7041) SmaI (7039) TspMI - XmaI (7037) SbfI (7018) HindIII (7002) AlwNI (6234) BspHI (5923) AhdI (5755) MfeI (4942) BstAPI (33) BsaAI (2736) EagI (3375) AfeI (3440) BstEII (3547) SacII (3777) BsmBI (3891) PflFI - Tth111I (3995) PpuMI * (4021) AgeI (4848) pPTR II 10,032 bp
NaeI  (9900)
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.
NgoMIV  (9898)
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.
KpnI  (7045)
1 site
G G T A C C C C A T G G
Acc65I  (7041)
1 site
G G T A C C C C A T G G
SmaI  (7039)
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  (7037)
1 site
C C C G G G G G G C C C
XmaI  (7037)
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.
SbfI  (7018)
1 site
C C T G C A G G G G A C G T C C
HindIII  (7002)
1 site
A A G C T T T T C G A A
AlwNI  (6234)
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  (5923)
1 site
T C A T G A A G T A C T
AhdI  (5755)
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.
MfeI  (4942)
1 site
C A A T T G G T T A A C
BstAPI  (33)
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.
BsaAI  (2736)
1 site
Y A C G T R R T G C A Y
EagI  (3375)
1 site
C G G C C G G C C G G C
AfeI  (3440)
1 site
A G C G C T T C G C G A
BstEII  (3547)
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.
SacII  (3777)
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.
BsmBI  (3891)
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.
PflFI  (3995)
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  (3995)
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.
PpuMI  (4021)
1 site
R G G W C C Y Y C C W G G R
* Blocked by Dcm methylation.
Sticky ends from different PpuMI sites may not be compatible.
AgeI  (4848)
1 site
A C C G G T T G G C C A
AMA1
7416 .. 2633  =  5250 bp
replication origin from Aspergillus nidulans (Aleksenko and Clutterbuck, 1996)
AMA1
7416 .. 2633  =  5250 bp
replication origin from Aspergillus nidulans (Aleksenko and Clutterbuck, 1996)
ptrA
2943 .. 4950  =  2008 bp
Product: pyrithiamine resistance gene of Aspergillus oryzae (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
ptrA
2943 .. 4950  =  2008 bp
Product: pyrithiamine resistance gene of Aspergillus oryzae (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
AmpR
4968 .. 5828  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4968 .. 5036  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4968 .. 5828  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5037 .. 5828  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4968 .. 5828  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
5999 .. 6587  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5999 .. 6587  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
lacZα
6985 .. 7311  =  327 bp
108 amino acids  =  12.4 kDa
Product: LacZα fragment of β-galactosidase
lacZα
6985 .. 7311  =  327 bp
108 amino acids  =  12.4 kDa
Product: LacZα fragment of β-galactosidase
lac promoter
6911 .. 6941  =  31 bp
   Segment 1:  -35  
   6911 .. 6916  =  6 bp
promoter for the E. coli lac operon
lac promoter
6911 .. 6941  =  31 bp
   Segment 2:  
   6917 .. 6934  =  18 bp
promoter for the E. coli lac operon
lac promoter
6911 .. 6941  =  31 bp
   Segment 3:  -10  
   6935 .. 6941  =  7 bp
promoter for the E. coli lac operon
lac promoter
6911 .. 6941  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
6949 .. 6965  =  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
6949 .. 6965  =  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).
ptrA
3187 .. 4228  =  1042 bp
327 amino acids  =  35.1 kDa
2 segments
Product: pyrithiamine resistance protein (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
ptrA
3187 .. 4228  =  1042 bp
327 amino acids  =  35.1 kDa
   Segment 2:  
   3187 .. 3833  =  647 bp
   215 amino acids  =  23.1 kDa
Product: pyrithiamine resistance protein (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
ptrA
3187 .. 4228  =  1042 bp
327 amino acids  =  35.1 kDa
   Segment 1:  
   3892 .. 4228  =  337 bp
   112 amino acids  =  12.0 kDa
Product: pyrithiamine resistance protein (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
ptrA
3187 .. 4228  =  1042 bp
327 amino acids  =  35.1 kDa
2 segments
Product: pyrithiamine resistance protein (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
MCS
7002 .. 7046  =  45 bp
multiple cloning site
MCS
7002 .. 7046  =  45 bp
multiple cloning site
M13 rev
6973 .. 6989  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
6973 .. 6989  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
7059 .. 7075  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
7059 .. 7075  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  6985 .. 7311  =  327 bp
ORF:  108 amino acids  =  12.4 kDa
ORF:  2954 .. 3178  =  225 bp
ORF:  74 amino acids  =  8.4 kDa
ORF:  4968 .. 5828  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3187 .. 3795  =  609 bp
ORF:  202 amino acids  =  21.9 kDa
ORF:  6912 .. 7268  =  357 bp
ORF:  118 amino acids  =  13.5 kDa
ORF:  3623 .. 3880  =  258 bp
ORF:  85 amino acids  =  9.7 kDa
ORF:  3884 .. 4228  =  345 bp
ORF:  114 amino acids  =  12.2 kDa
ORF:  5432 .. 5698  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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