pRI 909

Agrobacterium binary vector with a multiple cloning site (MCS), for plant cell transformation. The MCS is reversed in pRI 910.

Sequence Author: TaKaRa

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PpuMI (8573) PmeI (7884) M13 rev EcoRI (7674) SacI (7672) Eco53kI (7670) KpnI (7666) Acc65I (7662) SmaI (7660) TspMI - XmaI (7658) BamHI (7653) XbaI (7647) AccI (7642) SalI (7641) PstI - SbfI (7639) SphI (7633) HindIII (7623) M13 fwd FspI (6982) BsaAI (6780) ApaI (6151) PspOMI (6147) PciI (6031) NsiI (6028) MauBI (5916) TatI (5602) 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 909 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
EcoRI  (7674)
1 site
G A A T T C C T T A A G
SacI  (7672)
1 site
G A G C T C C T C G A G
Eco53kI  (7670)
1 site
G A G C T C C T C G A G
KpnI  (7666)
1 site
G G T A C C C C A T G G
Acc65I  (7662)
1 site
G G T A C C C C A T G G
SmaI  (7660)
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  (7658)
1 site
C C C G G G G G G C C C
XmaI  (7658)
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.
BamHI  (7653)
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.
XbaI  (7647)
1 site
T C T A G A A G A T C T
AccI  (7642)
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  (7641)
1 site
G T C G A C C A G C T G
PstI  (7639)
1 site
C T G C A G G A C G T C
SbfI  (7639)
1 site
C C T G C A G G G G A C G T C C
SphI  (7633)
1 site
G C A T G C C G T A C G
HindIII  (7623)
1 site
A A G C T T T T C G A A
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
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
7623 .. 7679  =  57 bp
pUC18/19 multiple cloning site
MCS
7623 .. 7679  =  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 fwd
7603 .. 7619  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
7603 .. 7619  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
7689 .. 7705  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
7689 .. 7705  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  2422 .. 3444  =  1023 bp
ORF:  340 amino acids  =  37.6 kDa
ORF:  4879 .. 5142  =  264 bp
ORF:  87 amino acids  =  9.7 kDa
ORF:  8140 .. 8370  =  231 bp
ORF:  76 amino acids  =  8.3 kDa
ORF:  1211 .. 2425  =  1215 bp
ORF:  404 amino acids  =  45.0 kDa
ORF:  6368 .. 6904  =  537 bp
ORF:  178 amino acids  =  19.9 kDa
ORF:  9161 .. 382  =  390 bp
ORF:  129 amino acids  =  14.4 kDa
ORF:  3597 .. 4814  =  1218 bp
ORF:  405 amino acids  =  44.3 kDa
ORF:  5142 .. 5633  =  492 bp
ORF:  163 amino acids  =  18.0 kDa
ORF:  6075 .. 6368  =  294 bp
ORF:  97 amino acids  =  10.7 kDa
ORF:  7527 .. 7766  =  240 bp
ORF:  79 amino acids  =  8.5 kDa
ORF:  3148 .. 3387  =  240 bp
ORF:  79 amino acids  =  9.2 kDa
ORF:  3721 .. 4092  =  372 bp
ORF:  123 amino acids  =  13.9 kDa
ORF:  6655 .. 7041  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  6051 .. 6398  =  348 bp
ORF:  115 amino acids  =  12.9 kDa
ORF:  8007 .. 8801  =  795 bp
ORF:  264 amino acids  =  31.0 kDa
ORF:  3467 .. 3742  =  276 bp
ORF:  91 amino acids  =  9.9 kDa
ORF:  4202 .. 4513  =  312 bp
ORF:  103 amino acids  =  10.9 kDa
ORF:  6419 .. 7213  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
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