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Plasmid Files

pRI 101-AN

Agrobacterium binary vector for foreign gene expression in dicotyledonous plants.

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pRI 101-AN Sequence and MappRI 101-AN.dna
Map and Sequence File   
Sequence Author:  TaKaRa
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 PmeI (9133) HindIII (9008) SphI (9006) PstI - SbfI (9000) BsaI (8913) ScaI (8783) PshAI (8340) XbaI (8143) AtADH 5'-UTR NdeI (8080) SalI (8070) SmaI (8066) TspMI - XmaI (8064) KpnI (8062) Acc65I (8058) BamHI (8052) EcoRI (8046) SacI (7987) Eco53kI (7985) M13 rev lac operator lac promoter CAP binding site NOS promoter FspI (6982) BsaAI (6780) ApaI (6151) PspOMI (6147) PspXI (5493) BbvCI (5292) ApaLI (310) BstEII (1838) BstAPI (2018) PflMI (2086) BclI * (2394) AgeI (4300) AvrII (5073) pRI 101-AN 10,417 bp
PmeI  (9133)
1 site
G T T T A A A C C A A A T T T G
HindIII  (9008)
1 site
A A G C T T T T C G A A
SphI  (9006)
1 site
G C A T G C C G T A C G
PstI  (9000)
1 site
C T G C A G G A C G T C
SbfI  (9000)
1 site
C C T G C A G G G G A C G T C C
BsaI  (8913)
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  (8783)
1 site
A G T A C T T C A T G A
PshAI  (8340)
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  (8143)
1 site
T C T A G A A G A T C T
NdeI  (8080)
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  (8070)
1 site
G T C G A C C A G C T G
SmaI  (8066)
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  (8064)
1 site
C C C G G G G G G C C C
XmaI  (8064)
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.
KpnI  (8062)
1 site
G G T A C C C C A T G G
Acc65I  (8058)
1 site
G G T A C C C C A T G G
BamHI  (8052)
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.
EcoRI  (8046)
1 site
G A A T T C C T T A A G
SacI  (7987)
1 site
G A G C T C C T C G A G
Eco53kI  (7985)
1 site
G A G C T C C T C G 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
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
9256 .. 10,050  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
9256 .. 10,050  =  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
8157 .. 8502  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
8157 .. 8502  =  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
NOS terminator
7720 .. 7972  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
7720 .. 7972  =  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
7983 .. 8084  =  102 bp
multiple cloning site
MCS
7983 .. 8084  =  102 bp
multiple cloning site
AtADH 5'-UTR
8085 .. 8142  =  58 bp
translational enhancer from the 5'-UTR of the
Arabidopsis thaliana alcohol dehydrogenase gene
(Sugio et al., 2008
AtADH 5'-UTR
8085 .. 8142  =  58 bp
translational enhancer from the 5'-UTR of the
Arabidopsis thaliana alcohol dehydrogenase gene
(Sugio et al., 2008
lac promoter
7620 .. 7650  =  31 bp
   Segment 1:  -35  
   7620 .. 7625  =  6 bp
promoter for the E. coli lac operon
lac promoter
7620 .. 7650  =  31 bp
   Segment 2:  
   7626 .. 7643  =  18 bp
promoter for the E. coli lac operon
lac promoter
7620 .. 7650  =  31 bp
   Segment 3:  -10  
   7644 .. 7650  =  7 bp
promoter for the E. coli lac operon
lac promoter
7620 .. 7650  =  31 bp
3 segments
promoter for the E. coli lac operon
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
9148 .. 9172  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
9148 .. 9172  =  25 bp
right border repeat from nopaline C58 T-DNA
CAP binding site
7584 .. 7605  =  22 bp
CAP binding activates transcription in the presence
of cAMP.
CAP binding site
7584 .. 7605  =  22 bp
CAP binding activates transcription in the presence
of cAMP.
lac operator
7658 .. 7674  =  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
7658 .. 7674  =  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).
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
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