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Plasmid Files

pCAMBIA0105.1R

Agrobacterium binary vector for plant transformation, with spectinomycin-resistance and GUSPlus™ genes.

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pCAMBIA0105.1R Sequence and MappCAMBIA0105.1R.dna
Map and Sequence File   
Sequence Author:  Cambia
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 NcoI (0) PshAI (9834) PvuII (9366) ApaI (9246) PspOMI (9242) XbaI (9236) PaeR7I - PspXI - XhoI (9224) PstI (9200) EcoRI (9191) BamHI (9160) SacI (9158) Eco53kI (9156) KpnI (9152) Acc65I (9148) HindIII (9142) lac operator AseI (9014) LB T-DNA repeat SacII (8682) BpmI (8113) BlpI (8051) HpaI (7524) BstZ17I (6451) AgeI (5961) EcoNI (5780) BglII (7) SspI (137) BstBI (556) BsrGI (652) StuI (730) SnaBI (769) Bpu10I (915) AleI (1554) ZraI (1610) AatII (1612) MluI (1696) BspHI * (1729) AfeI (1989) 6xHis PmlI (2048) AflII (2109) MauBI (2304) PasI (3797) AclI (4391) pCAMBIA0105.1R 10,033 bp
NcoI  (0)
1 site
C C A T G G G G T A C C
PshAI  (9834)
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.
PvuII  (9366)
1 site
C A G C T G G T C G A C
ApaI  (9246)
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  (9242)
1 site
G G G C C C C C C G G G
XbaI  (9236)
1 site
T C T A G A A G A T C T
PaeR7I  (9224)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (9224)
1 site
V C T C G A G B B G A G C T C V
XhoI  (9224)
1 site
C T C G A G G A G C T C
PstI  (9200)
1 site
C T G C A G G A C G T C
EcoRI  (9191)
1 site
G A A T T C C T T A A G
BamHI  (9160)
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.
SacI  (9158)
1 site
G A G C T C C T C G A G
Eco53kI  (9156)
1 site
G A G C T C C T C G A G
KpnI  (9152)
1 site
G G T A C C C C A T G G
Acc65I  (9148)
1 site
G G T A C C C C A T G G
HindIII  (9142)
1 site
A A G C T T T T C G A A
AseI  (9014)
1 site
A T T A A T T A A T T A
SacII  (8682)
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.
BpmI  (8113)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BlpI  (8051)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
HpaI  (7524)
1 site
G T T A A C C A A T T G
BstZ17I  (6451)
1 site
G T A T A C C A T A T G
AgeI  (5961)
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.
EcoNI  (5780)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BglII  (7)
1 site
A G A T C T T C T A G A
SspI  (137)
1 site
A A T A T T T T A T A A
BstBI  (556)
1 site
T T C G A A A A G C T T
BsrGI  (652)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
StuI  (730)
1 site
A G G C C T T C C G G A
SnaBI  (769)
1 site
T A C G T A A T G C A T
Bpu10I  (915)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
AleI  (1554)
1 site
C A C N N N N G T G G T G N N N N C A C
ZraI  (1610)
1 site
G A C G T C C T G C A G
AatII  (1612)
1 site
G A C G T C C T G C A G
MluI  (1696)
1 site
A C G C G T T G C G C A
BspHI  (1729)
1 site
T C A T G A A G T A C T
* Blocked by Dam methylation.
AfeI  (1989)
1 site
A G C G C T T C G C G A
PmlI  (2048)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AflII  (2109)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
MauBI  (2304)
1 site
C G C G C G C G G C G C G C G C
PasI  (3797)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
AclI  (4391)
1 site
A A C G T T T T G C A A
GUSPlus™
2 .. 2024  =  2023 bp
611 amino acids  =  69.8 kDa
2 segments
Product: β-glucuronidase
codon-optimized Staphylococcus gusA gene with a
catalase intron to ensure expression in plants but
not bacteria
GUSPlus™
2 .. 2024  =  2023 bp
611 amino acids  =  69.8 kDa
   Segment 1:  
   2 .. 16  =  15 bp
   5 amino acids  =  632.8 Da
Product: β-glucuronidase
codon-optimized Staphylococcus gusA gene with a
catalase intron to ensure expression in plants but
not bacteria
GUSPlus™
2 .. 2024  =  2023 bp
611 amino acids  =  69.8 kDa
   Segment 2:  
   207 .. 2024  =  1818 bp
   606 amino acids  =  69.2 kDa
Product: β-glucuronidase
codon-optimized Staphylococcus gusA gene with a
catalase intron to ensure expression in plants but
not bacteria
GUSPlus™
2 .. 2024  =  2023 bp
611 amino acids  =  69.8 kDa
2 segments
Product: β-glucuronidase
codon-optimized Staphylococcus gusA gene with a
catalase intron to ensure expression in plants but
not bacteria
pVS1 RepA
4736 .. 5809  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
4736 .. 5809  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
SmR
7571 .. 8362  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and
streptomycin
SmR
7571 .. 8362  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and
streptomycin
pVS1 StaA
3678 .. 4307  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
3678 .. 4307  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
6739 .. 7327  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6739 .. 7327  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CaMV 35S promoter
9673 .. 10,018  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
9673 .. 10,018  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
lacZα
9125 .. 9439  =  315 bp
104 amino acids  =  11.8 kDa
Product: LacZα fragment of β-galactosidase
lacZα
9125 .. 9439  =  315 bp
104 amino acids  =  11.8 kDa
Product: LacZα fragment of β-galactosidase
NOS terminator
2080 .. 2332  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
2080 .. 2332  =  253 bp
nopaline synthase terminator and poly(A) signal
pVS1 oriV
5875 .. 6069  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
5875 .. 6069  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
bom
6413 .. 6553  =  141 bp
basis of mobility region from pBR322
bom
6413 .. 6553  =  141 bp
basis of mobility region from pBR322
lac promoter
9051 .. 9081  =  31 bp
   Segment 1:  -35  
   9051 .. 9056  =  6 bp
promoter for the E. coli lac operon
lac promoter
9051 .. 9081  =  31 bp
   Segment 2:  
   9057 .. 9074  =  18 bp
promoter for the E. coli lac operon
lac promoter
9051 .. 9081  =  31 bp
   Segment 3:  -10  
   9075 .. 9081  =  7 bp
promoter for the E. coli lac operon
lac promoter
9051 .. 9081  =  31 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
2354 .. 2378  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
2354 .. 2378  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8923 .. 8947  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8923 .. 8947  =  25 bp
left border repeat from nopaline C58 T-DNA
6xHis
2031 .. 2048  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2031 .. 2048  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lac operator
9089 .. 9105  =  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
9089 .. 9105  =  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).
cat1 intron
17 .. 206  =  190 bp
castor bean catalase intron, modified
cat1 intron
17 .. 206  =  190 bp
castor bean catalase intron, modified
MCS
9142 .. 9247  =  106 bp
multiple cloning site
MCS
9142 .. 9247  =  106 bp
multiple cloning site
T7 promoter
9255 .. 9273  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
9255 .. 9273  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
9113 .. 9129  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
9113 .. 9129  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
9280 .. 9296  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
9280 .. 9296  =  17 bp
common sequencing primer, one of multiple similar
variants
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