pCAMBIA2300

Agrobacterium binary vector for plant transformation, with kanamycin-resistance genes.

Sequence Author: Cambia

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PmeI (8623) HindIII (8410) PstI - SbfI (8402) SalI (8392) XbaI (8386) BamHI (8380) SmaI (8377) KpnI - TspMI - XmaI (8375) Acc65I (8371) SacI (8369) Eco53kI (8367) EcoRI (8359) lac operator BstXI (8116) BglII (7316) NcoI (7301) BssHII (6771) RsrII (6653) PspXI (6451) BclI * (5936) SacII (5933) PsiI (5805) Bpu10I (5660) EcoO109I - PpuMI (5521) BspHI (5479) BlpI (5329) SspI (4983) NsiI (4945) BsaBI * (1331) PasI (1338) AclI (1932) BsiWI (2892) NheI (3008) BmtI (3012) EcoNI (3321) BsaI (3411) AgeI (3502) MreI - SgrAI (3790) BstZ17I (3992) pCAMBIA2300 8743 bp
PmeI  (8623)
1 site
G T T T A A A C C A A A T T T G
HindIII  (8410)
1 site
A A G C T T T T C G A A
PstI  (8402)
1 site
C T G C A G G A C G T C
SbfI  (8402)
1 site
C C T G C A G G G G A C G T C C
SalI  (8392)
1 site
G T C G A C C A G C T G
XbaI  (8386)
1 site
T C T A G A A G A T C T
BamHI  (8380)
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  (8377)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KpnI  (8375)
1 site
G G T A C C C C A T G G
TspMI  (8375)
1 site
C C C G G G G G G C C C
XmaI  (8375)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
Acc65I  (8371)
1 site
G G T A C C C C A T G G
SacI  (8369)
1 site
G A G C T C C T C G A G
Eco53kI  (8367)
1 site
G A G C T C C T C G A G
EcoRI  (8359)
1 site
G A A T T C C T T A A G
BstXI  (8116)
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.
BglII  (7316)
1 site
A G A T C T T C T A G A
NcoI  (7301)
1 site
C C A T G G G G T A C C
BssHII  (6771)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
RsrII  (6653)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
PspXI  (6451)
1 site
V C T C G A G B B G A G C T C V
BclI  (5936)
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.
SacII  (5933)
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.
PsiI  (5805)
1 site
T T A T A A A A T A T T
Bpu10I  (5660)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
EcoO109I  (5521)
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.
PpuMI  (5521)
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  (5479)
1 site
T C A T G A A G T A C T
BlpI  (5329)
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  (4983)
1 site
A A T A T T T T A T A A
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.
PasI  (1338)
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  (1932)
1 site
A A C G T T T T G C A A
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
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
MreI  (3790)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (3790)
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.
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
NeoR/KanR
6507 .. 7304  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
6507 .. 7304  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
KanR
4955 .. 5749  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
4955 .. 5749  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CaMV 35S promoter (enhanced)
7367 .. 8044  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CaMV 35S promoter (enhanced)
7367 .. 8044  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
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
lacZα
8345 .. 8578  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
8345 .. 8578  =  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
6276 .. 6450  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
6276 .. 6450  =  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
8271 .. 8301  =  31 bp
3 segments
   Segment 1:  -35  
   8271 .. 8276  =  6 bp
promoter for the E. coli lac operon
lac promoter
8271 .. 8301  =  31 bp
3 segments
   Segment 2:  
   8277 .. 8294  =  18 bp
promoter for the E. coli lac operon
lac promoter
8271 .. 8301  =  31 bp
3 segments
   Segment 3:  -10  
   8295 .. 8301  =  7 bp
promoter for the E. coli lac operon
lac promoter
8271 .. 8301  =  31 bp
3 segments
promoter for the E. coli lac operon
LB T-DNA repeat
6174 .. 6198  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6174 .. 6198  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8638 .. 8662  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8638 .. 8662  =  25 bp
right border repeat from nopaline C58 T-DNA
lac operator
8309 .. 8325  =  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
8309 .. 8325  =  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
8359 .. 8415  =  57 bp
pUC18/19 multiple cloning site
MCS
8359 .. 8415  =  57 bp
pUC18/19 multiple cloning site
M13 rev
8333 .. 8349  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
8333 .. 8349  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
8419 .. 8435  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
8419 .. 8435  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1219 .. 1848  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  8345 .. 8578  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  234 .. 920  =  687 bp
ORF:  228 amino acids  =  25.1 kDa
ORF:  2175 .. 3350  =  1176 bp
ORF:  391 amino acids  =  43.9 kDa
ORF:  5088 .. 5318  =  231 bp
ORF:  76 amino acids  =  8.3 kDa
ORF:  6738 .. 6992  =  255 bp
ORF:  84 amino acids  =  9.7 kDa
ORF:  266 .. 616  =  351 bp
ORF:  116 amino acids  =  12.6 kDa
ORF:  2123 .. 2380  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  4955 .. 5749  =  795 bp
ORF:  264 amino acids  =  31.0 kDa
ORF:  6743 .. 7129  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  685 .. 957  =  273 bp
ORF:  90 amino acids  =  9.9 kDa
ORF:  7471 .. 7788  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
ORF:  7798 .. 8256  =  459 bp
ORF:  152 amino acids  =  16.9 kDa
ORF:  957 .. 1271  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2145 .. 2717  =  573 bp
ORF:  190 amino acids  =  20.6 kDa
ORF:  6507 .. 7445  =  939 bp
ORF:  312 amino acids  =  34.6 kDa
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