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Plasmid Files

pCAMBIA1391Xb

Agrobacterium binary vector with hygromycin- and kanamycin-resistance and GUS genes plus the pUC9 MCS. For other reading frames, use pCAMBIA1391Xa or pCAMBIA1391Xc.

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pCAMBIA1391Xb Sequence and MappCAMBIA1391Xb.dna
Map and Sequence File   
Sequence Author:  Cambia
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 EcoRI (10,639) BamHI (10,629) SalI (10,623) PstI (10,621) HindIII (10,609) AscI (10,602) ApaI - BanII (10,599) PspOMI (10,595) BstXI (10,592) CaMV 35S promoter (enhanced) AatII (9751) ZraI (9749) AsiSI (9412) RsrII (9363) PspXI (8711) SacII (8193) PsiI (8065) Bpu10I (7920) EcoO109I - PpuMI (7781) BlpI (7589) BstZ17I (6252) PluTI (6057) SfoI (6055) NarI (6054) KasI (6053) MreI - SgrAI (6050) BsaI (5671) EcoNI (5581) FspI (5485) BspDI * - ClaI * (5348) AfeI (1) BglII (7) SpeI (14) MfeI (147) SnaBI (401) PvuII (1090) BstBI (1110) 6xHis PmlI (1849) BstEII (1859) AflII (1910) MauBI (2105) RB T-DNA repeat SphI (2264) PasI (3598) AclI (4192) pVS1 RepA BsiWI (5152) pCAMBIA1391Xb 10,647 bp
EcoRI  (10,639)
1 site
G A A T T C C T T A A G
BamHI  (10,629)
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  (10,623)
1 site
G T C G A C C A G C T G
PstI  (10,621)
1 site
C T G C A G G A C G T C
HindIII  (10,609)
1 site
A A G C T T T T C G A A
AscI  (10,602)
1 site
G G C G C G C C C C G C G C G G
ApaI  (10,599)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (10,599)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PspOMI  (10,595)
1 site
G G G C C C C C C G G G
BstXI  (10,592)
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  (9751)
1 site
G A C G T C C T G C A G
ZraI  (9749)
1 site
G A C G T C C T G C A G
AsiSI  (9412)
1 site
G C G A T C G C C G C T A G C G
RsrII  (9363)
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  (8711)
1 site
V C T C G A G B B G A G C T C V
SacII  (8193)
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  (8065)
1 site
T T A T A A A A T A T T
Bpu10I  (7920)
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.
EcoO109I  (7781)
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  (7781)
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.
BlpI  (7589)
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  (6252)
1 site
G T A T A C C A T A T G
PluTI  (6057)
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  (6055)
1 site
G G C G C C C C G C G G
NarI  (6054)
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  (6053)
1 site
G G C G C C C C G C G G
MreI  (6050)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (6050)
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  (5671)
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  (5581)
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.
FspI  (5485)
1 site
T G C G C A A C G C G T
BspDI  (5348)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (5348)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AfeI  (1)
1 site
A G C G C T T C G C G A
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
MfeI  (147)
1 site
C A A T T G G T T A A C
SnaBI  (401)
1 site
T A C G T A A T G C A T
PvuII  (1090)
1 site
C A G C T G G T C G A C
BstBI  (1110)
1 site
T T C G A A A A G C T T
PmlI  (1849)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BstEII  (1859)
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  (1910)
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  (2105)
1 site
C G C G C G C G G C G C G C G C
SphI  (2264)
1 site
G C A T G C C G T A C G
PasI  (3598)
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  (4192)
1 site
A A C G T T T T G C A A
BsiWI  (5152)
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.
GUS
20 .. 1825  =  1806 bp
602 amino acids  =  68.3 kDa
Product: β-glucuronidase
GUS
20 .. 1825  =  1806 bp
602 amino acids  =  68.3 kDa
Product: β-glucuronidase
pVS1 RepA
4537 .. 5610  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
4537 .. 5610  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
HygR
8750 .. 9775  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
8750 .. 9775  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
KanR
7215 .. 8009  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
7215 .. 8009  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CaMV 35S promoter (enhanced)
9843 .. 10,520  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
9843 .. 10,520  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
pVS1 StaA
3479 .. 4108  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
3479 .. 4108  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
6540 .. 7128  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6540 .. 7128  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
NOS terminator
1881 .. 2133  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
1881 .. 2133  =  253 bp
nopaline synthase terminator and poly(A) signal
pVS1 oriV
5676 .. 5870  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
5676 .. 5870  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
8536 .. 8710  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
8536 .. 8710  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
6214 .. 6354  =  141 bp
basis of mobility region from pBR322
bom
6214 .. 6354  =  141 bp
basis of mobility region from pBR322
MCS
10,609 .. 10,644  =  36 bp
multiple cloning site from pUC9
MCS
10,609 .. 10,644  =  36 bp
multiple cloning site from pUC9
RB T-DNA repeat
2155 .. 2179  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
2155 .. 2179  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8434 .. 8458  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8434 .. 8458  =  25 bp
left border repeat from nopaline C58 T-DNA
6xHis
1832 .. 1849  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1832 .. 1849  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
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