pCAMBIA1291Z

Agrobacterium binary vector for plant transformation, with hygromycin- and chloramphenicol-resistance and LacZ-GUS genes plus the pUC9 MCS.

Sequence Author: Cambia

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NcoI (0) AvrII (11,363) EcoRI (11,209) BamHI (11,199) SalI (11,193) PstI (11,191) HindIII (11,179) lac operator BstXI (10,946) AatII (10,105) ZraI (10,103) AsiSI (9766) RsrII (9717) PspXI (9065) HpaI (7527) BstZ17I (6454) PluTI (6259) SfoI (6257) NarI (6256) KasI (6255) BsaI (5873) EcoNI (5783) BglII (7) EcoO109I - PpuMI (54) AhdI (216) MfeI (346) SnaBI (600) BspHI (1100) BstBI (1309) 6xHis PmlI (2048) BstEII (2061) AflII (2112) MauBI (2307) SphI (2466) BsiWI (5354) BspDI * - ClaI * (5550) pCAMBIA1291Z 11,379 bp
NcoI  (0)
1 site
C C A T G G G G T A C C
AvrII  (11,363)
1 site
C C T A G G G G A T C C
EcoRI  (11,209)
1 site
G A A T T C C T T A A G
BamHI  (11,199)
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.
SalI  (11,193)
1 site
G T C G A C C A G C T G
PstI  (11,191)
1 site
C T G C A G G A C G T C
HindIII  (11,179)
1 site
A A G C T T T T C G A A
BstXI  (10,946)
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  (10,105)
1 site
G A C G T C C T G C A G
ZraI  (10,103)
1 site
G A C G T C C T G C A G
AsiSI  (9766)
1 site
G C G A T C G C C G C T A G C G
RsrII  (9717)
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
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
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
EcoO109I  (54)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PpuMI  (54)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
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
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
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
SphI  (2466)
1 site
G C A T G C C G T A C G
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.
lacZα
11,162 .. 11,359  =  198 bp
66 amino acids  =  7.4 kDa
Product: LacZα fragment of β-galactosidase
lacZα
11,162 .. 11,359  =  198 bp
66 amino acids  =  7.4 kDa
Product: LacZα fragment of β-galactosidase
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
HygR
9104 .. 10,129  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
9104 .. 10,129  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
CaMV 35S promoter (enhanced)
10,197 .. 10,874  =  678 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CaMV 35S promoter (enhanced)
10,197 .. 10,874  =  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
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
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
11,088 .. 11,118  =  31 bp
3 segments
   Segment 1:  -35  
   11,088 .. 11,093  =  6 bp
promoter for the E. coli lac operon
lac promoter
11,088 .. 11,118  =  31 bp
3 segments
   Segment 2:  
   11,094 .. 11,111  =  18 bp
promoter for the E. coli lac operon
lac promoter
11,088 .. 11,118  =  31 bp
3 segments
   Segment 3:  -10  
   11,112 .. 11,118  =  7 bp
promoter for the E. coli lac operon
lac promoter
11,088 .. 11,118  =  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
11,126 .. 11,142  =  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
11,126 .. 11,142  =  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
11,179 .. 11,214  =  36 bp
multiple cloning site from pUC9
MCS
11,179 .. 11,214  =  36 bp
multiple cloning site from pUC9
M13 rev
11,150 .. 11,166  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
11,150 .. 11,166  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
11,215 .. 11,231  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
11,215 .. 11,231  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  8413 .. 8655  =  243 bp
ORF:  80 amino acids  =  9.1 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:  9056 .. 9796  =  741 bp
ORF:  246 amino acids  =  27.0 kDa
ORF:  11,162 .. 28  =  246 bp
ORF:  81 amino acids  =  9.2 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:  11,089 .. 3  =  294 bp
ORF:  97 amino acids  =  10.7 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:  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:  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:  9104 .. 10,129  =  1026 bp
ORF:  341 amino acids  =  38.0 kDa
ORF:  10,301 .. 10,618  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
ORF:  10,628 .. 10,945  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
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