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Plasmid Files

pRI 201-ON

Agrobacterium binary vector for high-level expression of a foreign gene in monocotyledonous plants, with optional expression of a second gene.

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pRI 201-ON Sequence and MappRI 201-ON.dna
Map and Sequence File   
Sequence Author:  TaKaRa
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 PmeI (9191) HindIII (9066) SphI (9064) PstI - SbfI (9058) BsaI (8971) ScaI (8841) PshAI (8398) XbaI (8201) NdeI (8095) SalI (8062) SacI (7998) Eco53kI (7996) KpnI (7737) Acc65I (7733) NotI (7725) MCS 2 BamHI (7717) SmaI (7715) TspMI - XmaI (7713) NOS promoter FspI (6982) BsaAI (6780) ApaI (6151) PspOMI (6147) PciI (6031) NsiI (6028) MauBI (5916) PspXI (5493) BbvCI (5292) ApaLI (310) BstEII (1838) BstAPI (2018) PflMI (2086) BclI * (2394) AgeI (4300) AvrII (5073) pRI 201-ON 10,475 bp
PmeI  (9191)
1 site
G T T T A A A C C A A A T T T G
HindIII  (9066)
1 site
A A G C T T T T C G A A
SphI  (9064)
1 site
G C A T G C C G T A C G
PstI  (9058)
1 site
C T G C A G G A C G T C
SbfI  (9058)
1 site
C C T G C A G G G G A C G T C C
BsaI  (8971)
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  (8841)
1 site
A G T A C T T C A T G A
PshAI  (8398)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
XbaI  (8201)
1 site
T C T A G A A G A T C T
NdeI  (8095)
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.
SalI  (8062)
1 site
G T C G A C C A G C T G
SacI  (7998)
1 site
G A G C T C C T C G A G
Eco53kI  (7996)
1 site
G A G C T C C T C G A G
KpnI  (7737)
1 site
G G T A C C C C A T G G
Acc65I  (7733)
1 site
G G T A C C C C A T G G
NotI  (7725)
1 site
G C G G C C G C C G C C G G C G
BamHI  (7717)
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.
SmaI  (7715)
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  (7713)
1 site
C C C G G G G G G C C C
XmaI  (7713)
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.
FspI  (6982)
1 site
T G C G C A A C G C G T
BsaAI  (6780)
1 site
Y A C G T R R T G C A Y
ApaI  (6151)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (6147)
1 site
G G G C C C C C C G G G
PciI  (6031)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NsiI  (6028)
1 site
A T G C A T T A C G T A
MauBI  (5916)
1 site
C G C G C G C G G C G C G C G C
PspXI  (5493)
1 site
V C T C G A G B B G A G C T C V
BbvCI  (5292)
1 site
C C T C A G C G G A G T C G
ApaLI  (310)
1 site
G T G C A C C A C G T G
BstEII  (1838)
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.
BstAPI  (2018)
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.
PflMI  (2086)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BclI  (2394)
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.
AgeI  (4300)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AvrII  (5073)
1 site
C C T A G G G G A T C C
pRi replicator region
889 .. 5524  =  4636 bp
replicator region from Agrobacterium plasmid pRiA
4b
pRi replicator region
889 .. 5524  =  4636 bp
replicator region from Agrobacterium plasmid pRiA
4b
NeoR/KanR
6419 .. 7213  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
6419 .. 7213  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
KanR
9314 .. 10,108  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
9314 .. 10,108  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
57 .. 645  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
57 .. 645  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CaMV 35S promoter
8215 .. 8560  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
8215 .. 8560  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
5893 .. 6145  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
5893 .. 6145  =  253 bp
nopaline synthase terminator and poly(A) signal
HSP terminator
7744 .. 7992  =  249 bp
efficient transcription terminator from the
Arabidopsis thaliana heat shock protein 18.2 gene
(Nagaya et al., 2010)
HSP terminator
7744 .. 7992  =  249 bp
efficient transcription terminator from the
Arabidopsis thaliana heat shock protein 18.2 gene
(Nagaya et al., 2010)
NOS promoter
7270 .. 7449  =  180 bp
nopaline synthase promoter
NOS promoter
7270 .. 7449  =  180 bp
nopaline synthase promoter
OsADH 5'-UTR
8100 .. 8200  =  101 bp
translational enhancer from the 5'-UTR of the Oryza
sativa
alcohol dehydrogenase gene (Sugio et al.,
2008)
OsADH 5'-UTR
8100 .. 8200  =  101 bp
translational enhancer from the 5'-UTR of the Oryza
sativa
alcohol dehydrogenase gene (Sugio et al.,
2008)
MCS 1
8062 .. 8099  =  38 bp
multiple cloning site 1
MCS 1
8062 .. 8099  =  38 bp
multiple cloning site 1
MCS 2
7708 .. 7738  =  31 bp
multiple cloning site 2
MCS 2
7708 .. 7738  =  31 bp
multiple cloning site 2
LB T-DNA repeat
5531 .. 5555  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
5531 .. 5555  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
9206 .. 9230  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
9206 .. 9230  =  25 bp
right border repeat from nopaline C58 T-DNA
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