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Plasmid Files

pCAMBIA1302

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

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pCAMBIA1302 Sequence and MappCAMBIA1302.dna
Map and Sequence File   
Sequence Author:  Cambia
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 NcoI (0) HindIII (9788) PstI - SbfI (9780) SalI (9770) XbaI (9764) BamHI (9758) KpnI (9753) Acc65I (9749) BanII - SacI (9747) Eco53kI (9745) EcoRI (9737) lac operator BstXI (9494) AatII (8653) ZraI (8651) AsiSI (8314) RsrII (8265) PspXI (7613) SacII (7095) PsiI (6967) Bpu10I (6822) BspHI (6641) BlpI (6491) SspI (6145) PluTI (4959) BglII (7) SpeI (14) Bsu36I (279) BstBI (635) BstEII (761) MauBI (1007) RB T-DNA repeat PasI (2500) AclI (3094) BsiWI (4054) EcoNI (4483) MreI - SgrAI (4952) KasI (4955) NarI (4956) SfoI (4957) pCAMBIA1302 10,550 bp
NcoI  (0)
1 site
C C A T G G G G T A C C
HindIII  (9788)
1 site
A A G C T T T T C G A A
PstI  (9780)
1 site
C T G C A G G A C G T C
SbfI  (9780)
1 site
C C T G C A G G G G A C G T C C
SalI  (9770)
1 site
G T C G A C C A G C T G
XbaI  (9764)
1 site
T C T A G A A G A T C T
BamHI  (9758)
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.
KpnI  (9753)
1 site
G G T A C C C C A T G G
Acc65I  (9749)
1 site
G G T A C C C C A T G G
BanII  (9747)
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  (9747)
1 site
G A G C T C C T C G A G
Eco53kI  (9745)
1 site
G A G C T C C T C G A G
EcoRI  (9737)
1 site
G A A T T C C T T A A G
BstXI  (9494)
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.
AatII  (8653)
1 site
G A C G T C C T G C A G
ZraI  (8651)
1 site
G A C G T C C T G C A G
AsiSI  (8314)
1 site
G C G A T C G C C G C T A G C G
RsrII  (8265)
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  (7613)
1 site
V C T C G A G B B G A G C T C V
SacII  (7095)
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  (6967)
1 site
T T A T A A A A T A T T
Bpu10I  (6822)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
BspHI  (6641)
1 site
T C A T G A A G T A C T
BlpI  (6491)
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  (6145)
1 site
A A T A T T T T A T A A
PluTI  (4959)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
BglII  (7)
1 site
A G A T C T T C T A G A
SpeI  (14)
1 site
A C T A G T T G A T C A
Bsu36I  (279)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BstBI  (635)
1 site
T T C G A A A A G C T T
BstEII  (761)
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.
MauBI  (1007)
1 site
C G C G C G C G G C G C G C G C
PasI  (2500)
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  (3094)
1 site
A A C G T T T T G C A A
BsiWI  (4054)
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  (4483)
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.
MreI  (4952)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (4952)
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.
KasI  (4955)
1 site
G G C G C C C C G C G G
NarI  (4956)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (4957)
1 site
G G C G C C C C G C G G
pVS1 RepA
3439 .. 4512  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
3439 .. 4512  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
HygR
7652 .. 8677  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
7652 .. 8677  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
KanR
6117 .. 6911  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
6117 .. 6911  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
mgfp5
17 .. 727  =  711 bp
237 amino acids  =  26.7 kDa
Product: GFP with folding enhancement mutations
suitable for expression in plants due to removal of a
cryptic intron
mgfp5
17 .. 727  =  711 bp
237 amino acids  =  26.7 kDa
Product: GFP with folding enhancement mutations
suitable for expression in plants due to removal of a
cryptic intron
CaMV 35S promoter (enhanced)
8745 .. 9422  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
8745 .. 9422  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
pVS1 StaA
2381 .. 3010  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
2381 .. 3010  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
5442 .. 6030  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5442 .. 6030  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CaMV 35S promoter
10,190 .. 10,535  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
10,190 .. 10,535  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
783 .. 1035  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
783 .. 1035  =  253 bp
nopaline synthase terminator and poly(A) signal
lacZα
9723 .. 9956  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
9723 .. 9956  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
pVS1 oriV
4578 .. 4772  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
4578 .. 4772  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
7438 .. 7612  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
7438 .. 7612  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
5116 .. 5256  =  141 bp
basis of mobility region from pBR322
bom
5116 .. 5256  =  141 bp
basis of mobility region from pBR322
lac promoter
9649 .. 9679  =  31 bp
   Segment 1:  -35  
   9649 .. 9654  =  6 bp
promoter for the E. coli lac operon
lac promoter
9649 .. 9679  =  31 bp
   Segment 2:  
   9655 .. 9672  =  18 bp
promoter for the E. coli lac operon
lac promoter
9649 .. 9679  =  31 bp
   Segment 3:  -10  
   9673 .. 9679  =  7 bp
promoter for the E. coli lac operon
lac promoter
9649 .. 9679  =  31 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
1057 .. 1081  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
1057 .. 1081  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
7336 .. 7360  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
7336 .. 7360  =  25 bp
left border repeat from nopaline C58 T-DNA
6xHis
734 .. 751  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
734 .. 751  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lac operator
9687 .. 9703  =  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
9687 .. 9703  =  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
9737 .. 9793  =  57 bp
pUC18/19 multiple cloning site
MCS
9737 .. 9793  =  57 bp
pUC18/19 multiple cloning site
M13 rev
9711 .. 9727  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
9711 .. 9727  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
9797 .. 9813  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
9797 .. 9813  =  17 bp
common sequencing primer, one of multiple similar
variants
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