pCAMBIA2201

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

Sequence Author: Cambia

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lacZα HindIII (11,024) PstI - SbfI (11,016) SalI (11,006) XbaI (11,000) BamHI (10,994) SmaI (10,991) KpnI - TspMI - XmaI (10,989) Acc65I (10,985) SacI (10,983) Eco53kI (10,981) EcoRI (10,973) lac operator lac promoter BstXI (10,730) NeoR/KanR RsrII (9267) PspXI (9065) HpaI (7527) BstZ17I (6454) AgeI (5964) AhdI (216) SnaBI (600) BspHI (1100) BstBI (1309) 6xHis DraIII - PmlI (2048) BstEII (2061) AflII (2112) MauBI (2307) RB T-DNA repeat BsiWI (5354) EcoNI (5783) BsaI (5873) pCAMBIA2201 11,786 bp
HindIII  (11,024)
1 site
A A G C T T T T C G A A
PstI  (11,016)
1 site
C T G C A G G A C G T C
SbfI  (11,016)
1 site
C C T G C A G G G G A C G T C C
SalI  (11,006)
1 site
G T C G A C C A G C T G
XbaI  (11,000)
1 site
T C T A G A A G A T C T
BamHI  (10,994)
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  (10,991)
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  (10,989)
1 site
G G T A C C C C A T G G
TspMI  (10,989)
1 site
C C C G G G G G G C C C
XmaI  (10,989)
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  (10,985)
1 site
G G T A C C C C A T G G
SacI  (10,983)
1 site
G A G C T C C T C G A G
Eco53kI  (10,981)
1 site
G A G C T C C T C G A G
EcoRI  (10,973)
1 site
G A A T T C C T T A A G
BstXI  (10,730)
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.
RsrII  (9267)
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  (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
AgeI  (5964)
1 site
A C C G G T T G G C C 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
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
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.
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.
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.
GUS
2 .. 2024  =  2023 bp
611 amino acids  =  69.5 kDa
2 segments  =  1833 bp
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  =  1833 bp
   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
2 segments  =  1833 bp
   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  =  1833 bp
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.
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
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
NeoR/KanR
9121 .. 9918  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
9121 .. 9918  =  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)
9981 .. 10,658  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CaMV 35S promoter (enhanced)
9981 .. 10,658  =  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
CaMV 35S promoter
11,426 .. 11,771  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
CaMV 35S promoter
11,426 .. 11,771  =  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
10,959 .. 11,192  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
10,959 .. 11,192  =  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
10,885 .. 10,915  =  31 bp
3 segments
   Segment 1:  -35  
   10,885 .. 10,890  =  6 bp
promoter for the E. coli lac operon
lac promoter
10,885 .. 10,915  =  31 bp
3 segments
   Segment 2:  
   10,891 .. 10,908  =  18 bp
promoter for the E. coli lac operon
lac promoter
10,885 .. 10,915  =  31 bp
3 segments
   Segment 3:  -10  
   10,909 .. 10,915  =  7 bp
promoter for the E. coli lac operon
lac promoter
10,885 .. 10,915  =  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
lac operator
10,923 .. 10,939  =  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
10,923 .. 10,939  =  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
10,973 .. 11,029  =  57 bp
pUC18/19 multiple cloning site
MCS
10,973 .. 11,029  =  57 bp
pUC18/19 multiple cloning site
M13 rev
10,947 .. 10,963  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
10,947 .. 10,963  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
11,033 .. 11,049  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
11,033 .. 11,049  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  8413 .. 8655  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  9352 .. 9606  =  255 bp
ORF:  84 amino acids  =  9.7 kDa
ORF:  2696 .. 3382  =  687 bp
ORF:  228 amino acids  =  25.1 kDa
ORF:  4637 .. 5812  =  1176 bp
ORF:  391 amino acids  =  43.9 kDa
ORF:  8525 .. 8866  =  342 bp
ORF:  113 amino acids  =  12.1 kDa
ORF:  528 .. 2054  =  1527 bp
ORF:  508 amino acids  =  57.4 kDa
ORF:  3681 .. 4310  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  10,959 .. 11,192  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  11,244 .. 11,672  =  429 bp
ORF:  142 amino acids  =  15.8 kDa
ORF:  330 .. 791  =  462 bp
ORF:  153 amino acids  =  17.5 kDa
ORF:  1380 .. 1706  =  327 bp
ORF:  108 amino acids  =  12.8 kDa
ORF:  1758 .. 2153  =  396 bp
ORF:  131 amino acids  =  14.6 kDa
ORF:  3147 .. 3419  =  273 bp
ORF:  90 amino acids  =  9.9 kDa
ORF:  9357 .. 9743  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3419 .. 3733  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  4607 .. 5179  =  573 bp
ORF:  190 amino acids  =  20.6 kDa
ORF:  7601 .. 8401  =  801 bp
ORF:  266 amino acids  =  31.0 kDa
ORF:  10,085 .. 10,402  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
ORF:  10,412 .. 10,870  =  459 bp
ORF:  152 amino acids  =  16.9 kDa
ORF:  2728 .. 3078  =  351 bp
ORF:  116 amino acids  =  12.6 kDa
ORF:  4585 .. 4842  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  9121 .. 10,059  =  939 bp
ORF:  312 amino acids  =  34.6 kDa
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