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pCAMBIA3300

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

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pCAMBIA3300 Sequence and MappCAMBIA3300.dna
Map and Sequence File   
Sequence Author:  Cambia
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 lac operator BstXI (8186) CaMV 35S promoter (enhanced) AfeI (7226) ApaI (7170) PspOMI (7166) AarI (7053) FspAI (6989) PspXI (6835) BclI * (6320) PsiI (6189) Bpu10I (6044) PpuMI (5905) BspHI (5863) BlpI (5713) SspI (5367) NsiI (5329) BstZ17I (4376) PluTI (4181) EcoRI (0) Eco53kI (8) SacI (10) TspMI - XmaI (16) SmaI (18) BamHI (21) XbaI (27) PstI - SbfI (43) HindIII (51) PvuI (177) PmeI (264) BsaBI * (1715) PasI (1722) AclI (2316) BsiWI (3276) NheI (3392) BmtI (3396) BspDI * - ClaI * (3472) EcoNI (3705) BsaI (3795) AgeI (3886) MreI - SgrAI (4174) KasI (4177) NarI (4178) SfoI (4179) pCAMBIA3300 8429 bp
BstXI  (8186)
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  (7226)
1 site
A G C G C T T C G C G A
ApaI  (7170)
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  (7166)
1 site
G G G C C C C C C G G G
AarI  (7053)
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  (6989)
1 site
R T G C G C A Y Y A C G C G T R
PspXI  (6835)
1 site
V C T C G A G B B G A G C T C V
BclI  (6320)
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.
PsiI  (6189)
1 site
T T A T A A A A T A T T
Bpu10I  (6044)
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.
PpuMI  (5905)
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.
BspHI  (5863)
1 site
T C A T G A A G T A C T
BlpI  (5713)
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.
SspI  (5367)
1 site
A A T A T T T T A T A A
NsiI  (5329)
1 site
A T G C A T T A C G T A
BstZ17I  (4376)
1 site
G T A T A C C A T A T G
PluTI  (4181)
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.
EcoRI  (0)
1 site
G A A T T C C T T A A G
Eco53kI  (8)
1 site
G A G C T C C T C G A G
SacI  (10)
1 site
G A G C T C C T C G A G
TspMI  (16)
1 site
C C C G G G G G G C C C
XmaI  (16)
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.
SmaI  (18)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (21)
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  (27)
1 site
T C T A G A A G A T C T
PstI  (43)
1 site
C T G C A G G A C G T C
SbfI  (43)
1 site
C C T G C A G G G G A C G T C C
HindIII  (51)
1 site
A A G C T T T T C G A A
PvuI  (177)
1 site
C G A T C G G C T A G C
PmeI  (264)
1 site
G T T T A A A C C A A A T T T G
BsaBI  (1715)
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.
PasI  (1722)
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  (2316)
1 site
A A C G T T T T G C A A
BsiWI  (3276)
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  (3392)
1 site
G C T A G C C G A T C G
BmtI  (3396)
1 site
G C T A G C C G A T C G
BspDI  (3472)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3472)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EcoNI  (3705)
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  (3795)
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  (3886)
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.
MreI  (4174)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (4174)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
KasI  (4177)
1 site
G G C G C C C C G C G G
NarI  (4178)
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  (4179)
1 site
G G C G C C C C G C G G
pVS1 RepA
2661 .. 3734  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
2661 .. 3734  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
KanR
5339 .. 6133  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
5339 .. 6133  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CaMV 35S promoter (enhanced)
7437 .. 8114  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
7437 .. 8114  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
pVS1 StaA
1603 .. 2232  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
1603 .. 2232  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
4664 .. 5252  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4664 .. 5252  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
BlpR
6841 .. 7392  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
BlpR
6841 .. 7392  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
lacZα
8415 .. 219  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
8415 .. 219  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
pVS1 oriV
3800 .. 3994  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
3800 .. 3994  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
6660 .. 6834  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
6660 .. 6834  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
4338 .. 4478  =  141 bp
basis of mobility region from pBR322
bom
4338 .. 4478  =  141 bp
basis of mobility region from pBR322
lac promoter
8341 .. 8371  =  31 bp
   Segment 1:  -35  
   8341 .. 8346  =  6 bp
promoter for the E. coli lac operon
lac promoter
8341 .. 8371  =  31 bp
   Segment 2:  
   8347 .. 8364  =  18 bp
promoter for the E. coli lac operon
lac promoter
8341 .. 8371  =  31 bp
   Segment 3:  -10  
   8365 .. 8371  =  7 bp
promoter for the E. coli lac operon
lac promoter
8341 .. 8371  =  31 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
279 .. 303  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
279 .. 303  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6558 .. 6582  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6558 .. 6582  =  25 bp
left border repeat from nopaline C58 T-DNA
lac operator
8379 .. 8395  =  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
8379 .. 8395  =  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
8429 .. 56  =  57 bp
pUC18/19 multiple cloning site
MCS
8429 .. 56  =  57 bp
pUC18/19 multiple cloning site
M13 fwd
60 .. 76  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
60 .. 76  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
8403 .. 8419  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
8403 .. 8419  =  17 bp
common sequencing primer, one of multiple similar
variants
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