pCAMBIA2200

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

Sequence Author: Cambia

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PmeI (8775) HindIII (8562) PstI - SbfI (8554) SalI (8544) XbaI (8538) BamHI (8532) SmaI (8529) KpnI - TspMI - XmaI (8527) Acc65I (8523) SacI (8521) Eco53kI (8519) EcoRI (8511) lac operator BstXI (8268) EcoRV (7616) BglII (7468) NcoI (7453) BssHII (6923) RsrII (6805) PspXI (6603) BclI * (6088) BspEI (5584) SspI (5272) HpaI (5065) NsiI (4945) BsaBI * (1331) BsiWI (2892) NheI (3008) BmtI (3012) EcoNI (3321) BsaI (3411) AgeI (3502) BstZ17I (3992) pCAMBIA2200 8895 bp
PmeI  (8775)
1 site
G T T T A A A C C A A A T T T G
HindIII  (8562)
1 site
A A G C T T T T C G A A
PstI  (8554)
1 site
C T G C A G G A C G T C
SbfI  (8554)
1 site
C C T G C A G G G G A C G T C C
SalI  (8544)
1 site
G T C G A C C A G C T G
XbaI  (8538)
1 site
T C T A G A A G A T C T
BamHI  (8532)
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  (8529)
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  (8527)
1 site
G G T A C C C C A T G G
TspMI  (8527)
1 site
C C C G G G G G G C C C
XmaI  (8527)
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  (8523)
1 site
G G T A C C C C A T G G
SacI  (8521)
1 site
G A G C T C C T C G A G
Eco53kI  (8519)
1 site
G A G C T C C T C G A G
EcoRI  (8511)
1 site
G A A T T C C T T A A G
BstXI  (8268)
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  (7616)
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.
BglII  (7468)
1 site
A G A T C T T C T A G A
NcoI  (7453)
1 site
C C A T G G G G T A C C
BssHII  (6923)
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  (6805)
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  (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
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
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
6659 .. 7456  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
6659 .. 7456  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
CaMV 35S promoter (enhanced)
7519 .. 8196  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CaMV 35S promoter (enhanced)
7519 .. 8196  =  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
lacZα
8497 .. 8730  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
8497 .. 8730  =  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
8423 .. 8453  =  31 bp
3 segments
   Segment 1:  -35  
   8423 .. 8428  =  6 bp
promoter for the E. coli lac operon
lac promoter
8423 .. 8453  =  31 bp
3 segments
   Segment 2:  
   8429 .. 8446  =  18 bp
promoter for the E. coli lac operon
lac promoter
8423 .. 8453  =  31 bp
3 segments
   Segment 3:  -10  
   8447 .. 8453  =  7 bp
promoter for the E. coli lac operon
lac promoter
8423 .. 8453  =  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
8790 .. 8814  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8790 .. 8814  =  25 bp
right border repeat from nopaline C58 T-DNA
lac operator
8461 .. 8477  =  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
8461 .. 8477  =  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
8511 .. 8567  =  57 bp
pUC18/19 multiple cloning site
MCS
8511 .. 8567  =  57 bp
pUC18/19 multiple cloning site
M13 rev
8485 .. 8501  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
8485 .. 8501  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
8571 .. 8587  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
8571 .. 8587  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1219 .. 1848  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  8497 .. 8730  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  5951 .. 6193  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  6890 .. 7144  =  255 bp
ORF:  84 amino acids  =  9.7 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:  6063 .. 6404  =  342 bp
ORF:  113 amino acids  =  12.1 kDa
ORF:  685 .. 957  =  273 bp
ORF:  90 amino acids  =  9.9 kDa
ORF:  6895 .. 7281  =  387 bp
ORF:  128 amino acids  =  14.6 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:  5139 .. 5939  =  801 bp
ORF:  266 amino acids  =  31.0 kDa
ORF:  7623 .. 7940  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
ORF:  7950 .. 8408  =  459 bp
ORF:  152 amino acids  =  16.9 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:  6659 .. 7597  =  939 bp
ORF:  312 amino acids  =  34.6 kDa
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