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pCAMBIA1381Z

Agrobacterium binary vector for plant transformation, with hygromycin- and kanamycin-resistance and LacZ-GUS genes plus the pUC8 MCS.

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pCAMBIA1381Z Sequence and MappCAMBIA1381Z.dna
Map and Sequence File   
Sequence Author:  Cambia
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 NcoI (0) AvrII (11,211) HindIII (11,054) PstI (11,050) SalI (11,040) BamHI (11,034) EcoRI (11,024) lac operator BstXI (10,794) CaMV 35S promoter (enhanced) AatII (9953) ZraI (9951) AsiSI (9614) RsrII (9565) PspXI (8913) SacII (8395) PsiI (8267) Bpu10I (8122) BlpI (7791) BstZ17I (6454) PluTI (6259) SfoI (6257) NarI (6256) KasI (6255) MreI - SgrAI (6252) BsaI (5873) EcoNI (5783) BglII (7) AhdI (216) MfeI (346) SnaBI (600) BstBI (1309) 6xHis PmlI (2048) BstEII (2061) AflII (2112) MauBI (2307) RB T-DNA repeat SphI (2466) PasI (3800) AclI (4394) BsiWI (5354) BspDI * - ClaI * (5550) pCAMBIA1381Z 11,227 bp
NcoI  (0)
1 site
C C A T G G G G T A C C
AvrII  (11,211)
1 site
C C T A G G G G A T C C
HindIII  (11,054)
1 site
A A G C T T T T C G A A
PstI  (11,050)
1 site
C T G C A G G A C G T C
SalI  (11,040)
1 site
G T C G A C C A G C T G
BamHI  (11,034)
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.
EcoRI  (11,024)
1 site
G A A T T C C T T A A G
BstXI  (10,794)
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  (9953)
1 site
G A C G T C C T G C A G
ZraI  (9951)
1 site
G A C G T C C T G C A G
AsiSI  (9614)
1 site
G C G A T C G C C G C T A G C G
RsrII  (9565)
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

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.
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
PluTI  (6259)
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.
SfoI  (6257)
1 site
G G C G C C C C G C G G
NarI  (6256)
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.
KasI  (6255)
1 site
G G C G C C C C G C G 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.
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.
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.
BglII  (7)
1 site
A G A T C T T C T A G 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.
MfeI  (346)
1 site
C A A T T G G T T A A C
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
PmlI  (2048)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°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

The sticky ends produced by AflII are hard to ligate.
MauBI  (2307)
1 site
C G C G C G C G G C G C G C G C
SphI  (2466)
1 site
G C A T G C C G T A C G
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.
BspDI  (5550)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (5550)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
GUS
2 .. 2024  =  2023 bp
611 amino acids  =  69.5 kDa
2 segments
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
   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
   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
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.
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
HygR
8952 .. 9977  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
8952 .. 9977  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
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)
10,045 .. 10,722  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
10,045 .. 10,722  =  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
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
11,010 .. 11,207  =  198 bp
66 amino acids  =  7.5 kDa
Product: LacZα fragment of β-galactosidase
lacZα
11,010 .. 11,207  =  198 bp
66 amino acids  =  7.5 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
10,936 .. 10,966  =  31 bp
   Segment 1:  -35  
   10,936 .. 10,941  =  6 bp
promoter for the E. coli lac operon
lac promoter
10,936 .. 10,966  =  31 bp
   Segment 2:  
   10,942 .. 10,959  =  18 bp
promoter for the E. coli lac operon
lac promoter
10,936 .. 10,966  =  31 bp
   Segment 3:  -10  
   10,960 .. 10,966  =  7 bp
promoter for the E. coli lac operon
lac promoter
10,936 .. 10,966  =  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
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
lac operator
10,974 .. 10,990  =  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
10,974 .. 10,990  =  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
11,024 .. 11,059  =  36 bp
multiple cloning site from pUC8
MCS
11,024 .. 11,059  =  36 bp
multiple cloning site from pUC8
M13 rev
10,998 .. 11,014  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
10,998 .. 11,014  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
11,063 .. 11,079  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
11,063 .. 11,079  =  17 bp
common sequencing primer, one of multiple similar
variants
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