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Plasmid Files

pCAMBIA3301

Agrobacterium binary vector for plant transformation, with bialophos/phosphinothricin resistance and kanamycin resistance and GUS genes.

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pCAMBIA3301 Sequence and MappCAMBIA3301.dna
Map and Sequence File   
Sequence Author:  Cambia
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 NcoI (11,313) BspEI (10,978) HindIII (10,551) PstI - SbfI (10,543) XbaI (10,527) BamHI (10,521) SmaI (10,518) TspMI - XmaI (10,516) SacI (10,510) Eco53kI (10,508) EcoRI (10,500) lac operator BstXI (10,257) CaMV 35S promoter (enhanced) AfeI (9297) ApaI (9241) PspOMI (9237) FspAI (9060) PspXI (8906) PsiI (8260) Bpu10I (8115) BlpI (7784) BstZ17I (6447) PluTI (6252) SfoI (6250) NarI (6249) KasI (6248) MreI - SgrAI (6245) AgeI (5957) BsaI (5866) EcoNI (5776) BglII (0) AhdI (209) SnaBI (593) BstBI (1302) DraIII - PmlI (2041) BstEII (2054) AflII (2105) MauBI (2300) RB T-DNA repeat PasI (3793) AclI (4387) BsiWI (5347) BspDI * - ClaI * (5543) pCAMBIA3301 11,320 bp
NcoI  (11,313)
1 site
C C A T G G G G T A C C
BspEI  (10,978)
1 site
T C C G G A A G G C C T
HindIII  (10,551)
1 site
A A G C T T T T C G A A
PstI  (10,543)
1 site
C T G C A G G A C G T C
SbfI  (10,543)
1 site
C C T G C A G G G G A C G T C C
XbaI  (10,527)
1 site
T C T A G A A G A T C T
BamHI  (10,521)
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  (10,518)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (10,516)
1 site
C C C G G G G G G C C C
XmaI  (10,516)
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.
SacI  (10,510)
1 site
G A G C T C C T C G A G
Eco53kI  (10,508)
1 site
G A G C T C C T C G A G
EcoRI  (10,500)
1 site
G A A T T C C T T A A G
BstXI  (10,257)
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.
AfeI  (9297)
1 site
A G C G C T T C G C G A
ApaI  (9241)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (9237)
1 site
G G G C C C C C C G G G
FspAI  (9060)
1 site
R T G C G C A Y Y A C G C G T R
PspXI  (8906)
1 site
V C T C G A G B B G A G C T C V
PsiI  (8260)
1 site
T T A T A A A A T A T T
Bpu10I  (8115)
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  (7784)
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  (6447)
1 site
G T A T A C C A T A T G
PluTI  (6252)
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  (6250)
1 site
G G C G C C C C G C G G
NarI  (6249)
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  (6248)
1 site
G G C G C C C C G C G G
MreI  (6245)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (6245)
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  (5957)
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.
BsaI  (5866)
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  (5776)
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  (0)
1 site
A G A T C T T C T A G A
AhdI  (209)
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.
SnaBI  (593)
1 site
T A C G T A A T G C A T
BstBI  (1302)
1 site
T T C G A A A A G C T T
DraIII  (2041)
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.
PmlI  (2041)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BstEII  (2054)
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  (2105)
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  (2300)
1 site
C G C G C G C G G C G C G C G C
PasI  (3793)
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  (4387)
1 site
A A C G T T T T G C A A
BsiWI  (5347)
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  (5543)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (5543)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
GUS
11,315 .. 2017  =  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
11,315 .. 2017  =  2023 bp
611 amino acids  =  69.5 kDa
   Segment 1:  
   11,315 .. 9  =  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
11,315 .. 2017  =  2023 bp
611 amino acids  =  69.5 kDa
   Segment 2:  
   200 .. 2017  =  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
11,315 .. 2017  =  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
4732 .. 5805  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
4732 .. 5805  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
KanR
7410 .. 8204  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
7410 .. 8204  =  795 bp
264 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CaMV 35S promoter (enhanced)
9508 .. 10,185  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
9508 .. 10,185  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
pVS1 StaA
3674 .. 4303  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
3674 .. 4303  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
6735 .. 7323  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6735 .. 7323  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
BlpR
8912 .. 9463  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
BlpR
8912 .. 9463  =  552 bp
183 amino acids  =  20.6 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialophos or phosphinothricin
CaMV 35S promoter
10,953 .. 11,298  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
10,953 .. 11,298  =  346 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
2076 .. 2328  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
2076 .. 2328  =  253 bp
nopaline synthase terminator and poly(A) signal
lacZα
10,486 .. 10,719  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
10,486 .. 10,719  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
pVS1 oriV
5871 .. 6065  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
5871 .. 6065  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
8731 .. 8905  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
8731 .. 8905  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
6409 .. 6549  =  141 bp
basis of mobility region from pBR322
bom
6409 .. 6549  =  141 bp
basis of mobility region from pBR322
lac promoter
10,412 .. 10,442  =  31 bp
   Segment 1:  -35  
   10,412 .. 10,417  =  6 bp
promoter for the E. coli lac operon
lac promoter
10,412 .. 10,442  =  31 bp
   Segment 2:  
   10,418 .. 10,435  =  18 bp
promoter for the E. coli lac operon
lac promoter
10,412 .. 10,442  =  31 bp
   Segment 3:  -10  
   10,436 .. 10,442  =  7 bp
promoter for the E. coli lac operon
lac promoter
10,412 .. 10,442  =  31 bp
3 segments
promoter for the E. coli lac operon
RB T-DNA repeat
2350 .. 2374  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
2350 .. 2374  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8629 .. 8653  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
8629 .. 8653  =  25 bp
left border repeat from nopaline C58 T-DNA
6xHis
2024 .. 2041  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2024 .. 2041  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lac operator
10,450 .. 10,466  =  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
10,450 .. 10,466  =  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
10 .. 199  =  190 bp
castor bean catalase intron, modified
cat1 intron
10 .. 199  =  190 bp
castor bean catalase intron, modified
MCS
10,500 .. 10,556  =  57 bp
pUC18/19 multiple cloning site
MCS
10,500 .. 10,556  =  57 bp
pUC18/19 multiple cloning site
M13 rev
10,474 .. 10,490  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
10,474 .. 10,490  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
10,560 .. 10,576  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
10,560 .. 10,576  =  17 bp
common sequencing primer, one of multiple similar
variants
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