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pCAMBIA0305.2

Agrobacterium binary vector for plant transformation, with kanamycin-resistance and secreted GUSPlus™ genes.

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pCAMBIA0305.2 Sequence and MappCAMBIA0305.2.dna
Map and Sequence File   
Sequence Author:  Cambia
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 NcoI (0) PshAI (9538) PvuII (9070) HindIII (8975) PstI - SbfI (8967) SalI (8957) XbaI (8951) BamHI (8945) SmaI (8942) KpnI - TspMI - XmaI (8940) Acc65I (8936) BanII - SacI (8934) Eco53kI (8932) EcoRI (8924) lac operator AseI (8799) LB T-DNA repeat SacII (8467) PsiI (8339) BpmI (8012) BlpI (7863) BstZ17I (6526) MreI - SgrAI (6324) AgeI (6036) EcoNI (5855) GRP signal sequence BglII (82) SpeI (291) BstBI (631) BsrGI (727) StuI (805) SnaBI (844) AleI (1629) ZraI (1685) AatII (1687) MluI (1771) DraIII (1910) AfeI (2064) 6xHis PmlI (2123) BstEII (2133) AflII (2184) MauBI (2379) PasI (3872) AclI (4466) pCAMBIA0305.2 9737 bp
NcoI  (0)
1 site
C C A T G G G G T A C C
PshAI  (9538)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
PvuII  (9070)
1 site
C A G C T G G T C G A C
HindIII  (8975)
1 site
A A G C T T T T C G A A
PstI  (8967)
1 site
C T G C A G G A C G T C
SbfI  (8967)
1 site
C C T G C A G G G G A C G T C C
SalI  (8957)
1 site
G T C G A C C A G C T G
XbaI  (8951)
1 site
T C T A G A A G A T C T
BamHI  (8945)
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  (8942)
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  (8940)
1 site
G G T A C C C C A T G G
TspMI  (8940)
1 site
C C C G G G G G G C C C
XmaI  (8940)
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.
Acc65I  (8936)
1 site
G G T A C C C C A T G G
BanII  (8934)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (8934)
1 site
G A G C T C C T C G A G
Eco53kI  (8932)
1 site
G A G C T C C T C G A G
EcoRI  (8924)
1 site
G A A T T C C T T A A G
AseI  (8799)
1 site
A T T A A T T A A T T A
SacII  (8467)
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  (8339)
1 site
T T A T A A A A T A T T
BpmI  (8012)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BlpI  (7863)
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  (6526)
1 site
G T A T A C C A T A T G
MreI  (6324)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (6324)
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  (6036)
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.
EcoNI  (5855)
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.
BglII  (82)
1 site
A G A T C T T C T A G A
SpeI  (291)
1 site
A C T A G T T G A T C A
BstBI  (631)
1 site
T T C G A A A A G C T T
BsrGI  (727)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
StuI  (805)
1 site
A G G C C T T C C G G A
SnaBI  (844)
1 site
T A C G T A A T G C A T
AleI  (1629)
1 site
C A C N N N N G T G G T G N N N N C A C
ZraI  (1685)
1 site
G A C G T C C T G C A G
AatII  (1687)
1 site
G A C G T C C T G C A G
MluI  (1771)
1 site
A C G C G T T G C G C A
DraIII  (1910)
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.
AfeI  (2064)
1 site
A G C G C T T C G C G A
PmlI  (2123)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BstEII  (2133)
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  (2184)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
MauBI  (2379)
1 site
C G C G C G C G G C G C G C G C
PasI  (3872)
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  (4466)
1 site
A A C G T T T T G C A A
GUSPlus™
83 .. 2099  =  2017 bp
609 amino acids  =  69.6 kDa
2 segments
Product: β-glucuronidase
codon-optimized Staphylococcus gusA gene with a
catalase intron to ensure expression in plants but
not bacteria
GUSPlus™
83 .. 2099  =  2017 bp
609 amino acids  =  69.6 kDa
   Segment 1:  
   83 .. 91  =  9 bp
   3 amino acids  =  402.5 Da
Product: β-glucuronidase
codon-optimized Staphylococcus gusA gene with a
catalase intron to ensure expression in plants but
not bacteria
GUSPlus™
83 .. 2099  =  2017 bp
609 amino acids  =  69.6 kDa
   Segment 2:  
   282 .. 2099  =  1818 bp
   606 amino acids  =  69.2 kDa
Product: β-glucuronidase
codon-optimized Staphylococcus gusA gene with a
catalase intron to ensure expression in plants but
not bacteria
GUSPlus™
83 .. 2099  =  2017 bp
609 amino acids  =  69.6 kDa
2 segments
Product: β-glucuronidase
codon-optimized Staphylococcus gusA gene with a
catalase intron to ensure expression in plants but
not bacteria
pVS1 RepA
4811 .. 5884  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
4811 .. 5884  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
KanR
7489 .. 8283  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
7489 .. 8283  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
pVS1 StaA
3753 .. 4382  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
3753 .. 4382  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
6814 .. 7402  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6814 .. 7402  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CaMV 35S promoter
9377 .. 9722  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
9377 .. 9722  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
2155 .. 2407  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
2155 .. 2407  =  253 bp
nopaline synthase terminator and poly(A) signal
lacZα
8910 .. 9143  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
8910 .. 9143  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
pVS1 oriV
5950 .. 6144  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
5950 .. 6144  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
bom
6488 .. 6628  =  141 bp
basis of mobility region from pBR322
bom
6488 .. 6628  =  141 bp
basis of mobility region from pBR322
GRP signal sequence
2 .. 82  =  81 bp
27 amino acids  =  2.8 kDa
Product: signal sequence from Oryza sativa
glycine-rich cell wall protein
GRP signal sequence
2 .. 82  =  81 bp
27 amino acids  =  2.8 kDa
Product: signal sequence from Oryza sativa
glycine-rich cell wall protein
lac promoter
8836 .. 8866  =  31 bp
   Segment 1:  -35  
   8836 .. 8841  =  6 bp
promoter for the E. coli lac operon
lac promoter
8836 .. 8866  =  31 bp
   Segment 2:  
   8842 .. 8859  =  18 bp
promoter for the E. coli lac operon
lac promoter
8836 .. 8866  =  31 bp
   Segment 3:  -10  
   8860 .. 8866  =  7 bp
promoter for the E. coli lac operon
lac promoter
8836 .. 8866  =  31 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
2429 .. 2453  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
2429 .. 2453  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8708 .. 8732  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8708 .. 8732  =  25 bp
left border repeat from nopaline C58 T-DNA
6xHis
2106 .. 2123  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2106 .. 2123  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lac operator
8874 .. 8890  =  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
8874 .. 8890  =  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
92 .. 281  =  190 bp
castor bean catalase intron, modified
cat1 intron
92 .. 281  =  190 bp
castor bean catalase intron, modified
MCS
8924 .. 8980  =  57 bp
pUC18/19 multiple cloning site
MCS
8924 .. 8980  =  57 bp
pUC18/19 multiple cloning site
M13 rev
8898 .. 8914  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
8898 .. 8914  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
8984 .. 9000  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
8984 .. 9000  =  17 bp
common sequencing primer, one of multiple similar
variants
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