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Plasmid Files

pCAMBIA3201

Agrobacterium binary vector for plant transformation, with bialophos/phosphinothricin resistance and chloramphenicol resistance and GUS genes.

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pCAMBIA3201 Sequence and MappCAMBIA3201.dna
Map and Sequence File   
Sequence Author:  Cambia
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 NcoI (0) HindIII (10,710) PstI - SbfI (10,702) XbaI (10,686) BamHI (10,680) SmaI (10,677) TspMI - XmaI (10,675) SacI (10,669) Eco53kI (10,667) EcoRI (10,659) lac operator BstXI (10,416) AfeI (9456) ApaI (9400) PspOMI (9396) FspAI (9219) SacII (9189) PspXI (9065) HpaI (7527) BstZ17I (6454) PluTI (6259) SfoI (6257) NarI (6256) KasI (6255) AgeI (5964) BsaI (5873) EcoNI (5783) BglII (7) AhdI (216) SnaBI (600) BspHI (1100) BstBI (1309) 6xHis DraIII - PmlI (2048) BstEII (2061) AflII (2112) MauBI (2307) BsiWI (5354) BspDI * - ClaI * (5550) pCAMBIA3201 11,472 bp
NcoI  (0)
1 site
C C A T G G G G T A C C
HindIII  (10,710)
1 site
A A G C T T T T C G A A
PstI  (10,702)
1 site
C T G C A G G A C G T C
SbfI  (10,702)
1 site
C C T G C A G G G G A C G T C C
XbaI  (10,686)
1 site
T C T A G A A G A T C T
BamHI  (10,680)
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  (10,677)
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  (10,675)
1 site
C C C G G G G G G C C C
XmaI  (10,675)
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.
SacI  (10,669)
1 site
G A G C T C C T C G A G
Eco53kI  (10,667)
1 site
G A G C T C C T C G A G
EcoRI  (10,659)
1 site
G A A T T C C T T A A G
BstXI  (10,416)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
AfeI  (9456)
1 site
A G C G C T T C G C G A
ApaI  (9400)
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  (9396)
1 site
G G G C C C C C C G G G
FspAI  (9219)
1 site
R T G C G C A Y Y A C G C G T R
SacII  (9189)
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.
PspXI  (9065)
1 site
V C T C G A G B B G A G C T C V
HpaI  (7527)
1 site
G T T A A C C A A T T G
BstZ17I  (6454)
1 site
G T A T A C C A T A T G
PluTI  (6259)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (6257)
1 site
G G C G C C C C G C G G
NarI  (6256)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
KasI  (6255)
1 site
G G C G C C C C G C G G
AgeI  (5964)
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.
BsaI  (5873)
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.
EcoNI  (5783)
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
AhdI  (216)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
SnaBI  (600)
1 site
T A C G T A A T G C A T
BspHI  (1100)
1 site
T C A T G A A G T A C T
BstBI  (1309)
1 site
T T C G A A A A G C T T
DraIII  (2048)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
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.
BstEII  (2061)
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.
AflII  (2112)
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  (2307)
1 site
C G C G C G C G G C G C G C G C
BsiWI  (5354)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BspDI  (5550)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (5550)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
GUS
2 .. 2024  =  2023 bp
611 amino acids  =  69.5 kDa
2 segments
Product: β-glucuronidase
This version of the gusA gene has a 5' extension
with a catalase intron to ensure expression in plants
but not bacteria.
GUS
2 .. 2024  =  2023 bp
611 amino acids  =  69.5 kDa
   Segment 1:  
   2 .. 16  =  15 bp
   5 amino acids  =  632.8 Da
Product: β-glucuronidase
This version of the gusA gene has a 5' extension
with a catalase intron to ensure expression in plants
but not bacteria.
GUS
2 .. 2024  =  2023 bp
611 amino acids  =  69.5 kDa
   Segment 2:  
   207 .. 2024  =  1818 bp
   606 amino acids  =  68.9 kDa
Product: β-glucuronidase
This version of the gusA gene has a 5' extension
with a catalase intron to ensure expression in plants
but not bacteria.
GUS
2 .. 2024  =  2023 bp
611 amino acids  =  69.5 kDa
2 segments
Product: β-glucuronidase
This version of the gusA gene has a 5' extension
with a catalase intron to ensure expression in plants
but not bacteria.
pVS1 RepA
4739 .. 5812  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
4739 .. 5812  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
CaMV 35S promoter (enhanced)
9667 .. 10,344  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
9667 .. 10,344  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CmR
7601 .. 8260  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
7601 .. 8260  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
pVS1 StaA
3681 .. 4310  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
3681 .. 4310  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
6742 .. 7330  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6742 .. 7330  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
BlpR
9071 .. 9622  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
BlpR
9071 .. 9622  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
CaMV 35S promoter
11,112 .. 11,457  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
11,112 .. 11,457  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
2083 .. 2335  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
2083 .. 2335  =  253 bp
nopaline synthase terminator and poly(A) signal
lacZα
10,645 .. 10,878  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
10,645 .. 10,878  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
pVS1 oriV
5878 .. 6072  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
5878 .. 6072  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
8890 .. 9064  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
8890 .. 9064  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
6416 .. 6556  =  141 bp
basis of mobility region from pBR322
bom
6416 .. 6556  =  141 bp
basis of mobility region from pBR322
lac promoter
10,571 .. 10,601  =  31 bp
   Segment 1:  -35  
   10,571 .. 10,576  =  6 bp
promoter for the E. coli lac operon
lac promoter
10,571 .. 10,601  =  31 bp
   Segment 2:  
   10,577 .. 10,594  =  18 bp
promoter for the E. coli lac operon
lac promoter
10,571 .. 10,601  =  31 bp
   Segment 3:  -10  
   10,595 .. 10,601  =  7 bp
promoter for the E. coli lac operon
lac promoter
10,571 .. 10,601  =  31 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
2357 .. 2381  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
2357 .. 2381  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8788 .. 8812  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8788 .. 8812  =  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
10,609 .. 10,625  =  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
10,609 .. 10,625  =  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
10,659 .. 10,715  =  57 bp
pUC18/19 multiple cloning site
MCS
10,659 .. 10,715  =  57 bp
pUC18/19 multiple cloning site
M13 rev
10,633 .. 10,649  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
10,633 .. 10,649  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
10,719 .. 10,735  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
10,719 .. 10,735  =  17 bp
common sequencing primer, one of multiple similar
variants
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