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Plasmid Files

pRI 910

Agrobacterium binary vector with a multiple cloning site (MCS), for plant cell transformation. The MCS is reversed in pRI 909.

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pRI 910 Sequence and MappRI 910.dna
Map and Sequence File   
Sequence Author:  TaKaRa
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 PpuMI (8573) PmeI (7884) M13 fwd HindIII (7759) SphI (7757) PstI - SbfI (7751) AccI (7742) SalI (7741) XbaI (7735) BamHI (7729) SmaI (7726) KpnI - TspMI - XmaI (7724) Acc65I (7720) SacI (7718) Eco53kI (7716) EcoRI (7708) M13 rev FspI (6982) NeoR/KanR BsaAI (6780) ApaI (6151) PspOMI (6147) PciI (6031) NsiI (6028) MauBI (5916) TatI (5602) LB T-DNA repeat PspXI (5493) BbvCI (5292) AvrII (5073) ApaLI (310) MfeI (1473) BstEII (1838) BstAPI (2018) PflMI (2086) BclI * (2394) BmgBI (3159) BspEI (3940) AgeI (4300) pRI 910 9168 bp
PpuMI  (8573)
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.
PmeI  (7884)
1 site
G T T T A A A C C A A A T T T G
HindIII  (7759)
1 site
A A G C T T T T C G A A
SphI  (7757)
1 site
G C A T G C C G T A C G
PstI  (7751)
1 site
C T G C A G G A C G T C
SbfI  (7751)
1 site
C C T G C A G G G G A C G T C C
AccI  (7742)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
SalI  (7741)
1 site
G T C G A C C A G C T G
XbaI  (7735)
1 site
T C T A G A A G A T C T
BamHI  (7729)
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  (7726)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KpnI  (7724)
1 site
G G T A C C C C A T G G
TspMI  (7724)
1 site
C C C G G G G G G C C C
XmaI  (7724)
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.
Acc65I  (7720)
1 site
G G T A C C C C A T G G
SacI  (7718)
1 site
G A G C T C C T C G A G
Eco53kI  (7716)
1 site
G A G C T C C T C G A G
EcoRI  (7708)
1 site
G A A T T C C T T A A G
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
TatI  (5602)
1 site
W G T A C W W C A T G W
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
AvrII  (5073)
1 site
C C T A G G G G A T C C
ApaLI  (310)
1 site
G T G C A C C A C G T G
MfeI  (1473)
1 site
C A A T T G G T T A A C
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.
BmgBI  (3159)
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.
BspEI  (3940)
1 site
T C C G G A A G G C C T
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.
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
8007 .. 8801  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
8007 .. 8801  =  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
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
NOS promoter
7270 .. 7449  =  180 bp
nopaline synthase promoter
NOS promoter
7270 .. 7449  =  180 bp
nopaline synthase promoter
MCS
7708 .. 7764  =  57 bp
pUC18/19 multiple cloning site
MCS
7708 .. 7764  =  57 bp
pUC18/19 multiple cloning site
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
7899 .. 7923  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
7899 .. 7923  =  25 bp
right border repeat from nopaline C58 T-DNA
M13 rev
7682 .. 7698  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
7682 .. 7698  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
7768 .. 7784  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
7768 .. 7784  =  17 bp
common sequencing primer, one of multiple similar
variants
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