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pCAMBIA3200

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

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pCAMBIA3200 Sequence and MappCAMBIA3200.dna
Map and Sequence File   
Sequence Author:  Cambia
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 PmeI (8461) PvuI (8374) HindIII (8248) PstI - SbfI (8240) XbaI (8224) BamHI (8218) SmaI (8215) TspMI - XmaI (8213) SacI (8207) Eco53kI (8205) EcoRI (8197) lac operator BstXI (7954) CaMV 35S promoter (enhanced) EcoRV (7302) AfeI (6994) ApaI (6938) EcoO109I - PspOMI (6934) AarI (6821) FspAI (6757) SacII (6727) PspXI (6603) BclI * (6088) BspEI (5584) SspI (5272) HpaI (5065) NsiI (4945) BsaBI * (1331) BsiWI (2892) NheI (3008) BmtI (3012) BspDI * - ClaI * (3088) EcoNI (3321) BsaI (3411) AgeI (3502) KasI (3793) NarI (3794) SfoI (3795) PluTI (3797) BstZ17I (3992) pCAMBIA3200 8581 bp
PmeI  (8461)
1 site
G T T T A A A C C A A A T T T G
PvuI  (8374)
1 site
C G A T C G G C T A G C
HindIII  (8248)
1 site
A A G C T T T T C G A A
PstI  (8240)
1 site
C T G C A G G A C G T C
SbfI  (8240)
1 site
C C T G C A G G G G A C G T C C
XbaI  (8224)
1 site
T C T A G A A G A T C T
BamHI  (8218)
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  (8215)
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  (8213)
1 site
C C C G G G G G G C C C
XmaI  (8213)
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  (8207)
1 site
G A G C T C C T C G A G
Eco53kI  (8205)
1 site
G A G C T C C T C G A G
EcoRI  (8197)
1 site
G A A T T C C T T A A G
BstXI  (7954)
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.
EcoRV  (7302)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
AfeI  (6994)
1 site
A G C G C T T C G C G A
ApaI  (6938)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoO109I  (6934)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PspOMI  (6934)
1 site
G G G C C C C C C G G G
AarI  (6821)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
FspAI  (6757)
1 site
R T G C G C A Y Y A C G C G T R
SacII  (6727)
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  (6603)
1 site
V C T C G A G B B G A G C T C V
BclI  (6088)
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.
BspEI  (5584)
1 site
T C C G G A A G G C C T
SspI  (5272)
1 site
A A T A T T T T A T A A
HpaI  (5065)
1 site
G T T A A C C A A T T G
NsiI  (4945)
1 site
A T G C A T T A C G T A
BsaBI  (1331)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BsiWI  (2892)
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.
NheI  (3008)
1 site
G C T A G C C G A T C G
BmtI  (3012)
1 site
G C T A G C C G A T C G
BspDI  (3088)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3088)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EcoNI  (3321)
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.
BsaI  (3411)
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.
AgeI  (3502)
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.
KasI  (3793)
1 site
G G C G C C C C G C G G
NarI  (3794)
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.
SfoI  (3795)
1 site
G G C G C C C C G C G G
PluTI  (3797)
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.
BstZ17I  (3992)
1 site
G T A T A C C A T A T G
pVS1 RepA
2277 .. 3350  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
2277 .. 3350  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
CaMV 35S promoter (enhanced)
7205 .. 7882  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
7205 .. 7882  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CmR
5139 .. 5798  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
5139 .. 5798  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
pVS1 StaA
1219 .. 1848  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
1219 .. 1848  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
4280 .. 4868  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4280 .. 4868  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
BlpR
6609 .. 7160  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
BlpR
6609 .. 7160  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
lacZα
8183 .. 8416  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
8183 .. 8416  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
pVS1 oriV
3416 .. 3610  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
3416 .. 3610  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
6428 .. 6602  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
6428 .. 6602  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
3954 .. 4094  =  141 bp
basis of mobility region from pBR322
bom
3954 .. 4094  =  141 bp
basis of mobility region from pBR322
lac promoter
8109 .. 8139  =  31 bp
   Segment 1:  -35  
   8109 .. 8114  =  6 bp
promoter for the E. coli lac operon
lac promoter
8109 .. 8139  =  31 bp
   Segment 2:  
   8115 .. 8132  =  18 bp
promoter for the E. coli lac operon
lac promoter
8109 .. 8139  =  31 bp
   Segment 3:  -10  
   8133 .. 8139  =  7 bp
promoter for the E. coli lac operon
lac promoter
8109 .. 8139  =  31 bp
3 segments
promoter for the E. coli lac operon
LB T-DNA repeat
6326 .. 6350  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6326 .. 6350  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8476 .. 8500  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8476 .. 8500  =  25 bp
right border repeat from nopaline C58 T-DNA
lac operator
8147 .. 8163  =  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
8147 .. 8163  =  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).
MCS
8197 .. 8253  =  57 bp
pUC18/19 multiple cloning site
MCS
8197 .. 8253  =  57 bp
pUC18/19 multiple cloning site
M13 rev
8171 .. 8187  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
8171 .. 8187  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
8257 .. 8273  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
8257 .. 8273  =  17 bp
common sequencing primer, one of multiple similar
variants
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