pCAMBIA2301

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

Sequence Author: Cambia

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BspEI (11,299) HindIII (10,872) PstI - SbfI (10,864) SalI (10,854) XbaI (10,848) BamHI (10,842) SmaI (10,839) KpnI - TspMI - XmaI (10,837) Acc65I (10,833) SacI (10,831) Eco53kI (10,829) EcoRI (10,821) lac operator BstXI (10,578) RsrII (9115) PspXI (8913) SacII (8395) PsiI (8267) Bpu10I (8122) BlpI (7791) BstZ17I (6454) MreI - SgrAI (6252) AgeI (5964) BsaI (5873) AhdI (216) SnaBI (600) BstBI (1309) DraIII - PmlI (2048) BstEII (2061) AflII (2112) MauBI (2307) RB T-DNA repeat PasI (3800) AclI (4394) BsiWI (5354) EcoNI (5783) pCAMBIA2301 11,634 bp
BspEI  (11,299)
1 site
T C C G G A A G G C C T
HindIII  (10,872)
1 site
A A G C T T T T C G A A
PstI  (10,864)
1 site
C T G C A G G A C G T C
SbfI  (10,864)
1 site
C C T G C A G G G G A C G T C C
SalI  (10,854)
1 site
G T C G A C C A G C T G
XbaI  (10,848)
1 site
T C T A G A A G A T C T
BamHI  (10,842)
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  (10,839)
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  (10,837)
1 site
G G T A C C C C A T G G
TspMI  (10,837)
1 site
C C C G G G G G G C C C
XmaI  (10,837)
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  (10,833)
1 site
G G T A C C C C A T G G
SacI  (10,831)
1 site
G A G C T C C T C G A G
Eco53kI  (10,829)
1 site
G A G C T C C T C G A G
EcoRI  (10,821)
1 site
G A A T T C C T T A A G
BstXI  (10,578)
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  (9115)
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  (8913)
1 site
V C T C G A G B B G A G C T C V
SacII  (8395)
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  (8267)
1 site
T T A T A A A A T A T T
Bpu10I  (8122)
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.
BlpI  (7791)
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.
BstZ17I  (6454)
1 site
G T A T A C C A T A T G
MreI  (6252)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (6252)
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.
AgeI  (5964)
1 site
A C C G G T T G G C C A
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.
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
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
PasI  (3800)
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  (4394)
1 site
A A C G T T T T G C A A
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.
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
8969 .. 9766  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
8969 .. 9766  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
KanR
7417 .. 8211  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
7417 .. 8211  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CaMV 35S promoter (enhanced)
9829 .. 10,506  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CaMV 35S promoter (enhanced)
9829 .. 10,506  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
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
11,274 .. 11,619  =  346 bp
strong constitutive promoter from cauliflower mosaic virus
CaMV 35S promoter
11,274 .. 11,619  =  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
10,807 .. 11,040  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
10,807 .. 11,040  =  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
8738 .. 8912  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
8738 .. 8912  =  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
10,733 .. 10,763  =  31 bp
3 segments
   Segment 1:  -35  
   10,733 .. 10,738  =  6 bp
promoter for the E. coli lac operon
lac promoter
10,733 .. 10,763  =  31 bp
3 segments
   Segment 2:  
   10,739 .. 10,756  =  18 bp
promoter for the E. coli lac operon
lac promoter
10,733 .. 10,763  =  31 bp
3 segments
   Segment 3:  -10  
   10,757 .. 10,763  =  7 bp
promoter for the E. coli lac operon
lac promoter
10,733 .. 10,763  =  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
8636 .. 8660  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8636 .. 8660  =  25 bp
left border repeat from nopaline C58 T-DNA
lac operator
10,771 .. 10,787  =  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
10,771 .. 10,787  =  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
10,821 .. 10,877  =  57 bp
pUC18/19 multiple cloning site
MCS
10,821 .. 10,877  =  57 bp
pUC18/19 multiple cloning site
M13 rev
10,795 .. 10,811  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
10,795 .. 10,811  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
10,881 .. 10,897  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
10,881 .. 10,897  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  10,807 .. 11,040  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  11,092 .. 11,520  =  429 bp
ORF:  142 amino acids  =  15.8 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:  7550 .. 7780  =  231 bp
ORF:  76 amino acids  =  8.3 kDa
ORF:  9200 .. 9454  =  255 bp
ORF:  84 amino acids  =  9.7 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:  2728 .. 3078  =  351 bp
ORF:  116 amino acids  =  12.6 kDa
ORF:  4585 .. 4842  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  7417 .. 8211  =  795 bp
ORF:  264 amino acids  =  31.0 kDa
ORF:  9205 .. 9591  =  387 bp
ORF:  128 amino acids  =  14.6 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:  9933 .. 10,250  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
ORF:  10,260 .. 10,718  =  459 bp
ORF:  152 amino acids  =  16.9 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:  8969 .. 9907  =  939 bp
ORF:  312 amino acids  =  34.6 kDa
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