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Plasmid Files

pPZP212

Agrobacterium binary vector for plant transformation, with spectinomycin- and kanamycin-resistance genes. The MCS is reversed in pPZP211.

 
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 PmeI (8863) lac operator EcoRI (8584) SacI (8582) Eco53kI (8580) KpnI (8576) Acc65I (8572) SmaI (8570) TspMI - XmaI (8568) BamHI (8563) XbaI (8557) SalI (8551) PstI - SbfI (8549) HindIII (8533) BstXI (8356) EcoRV (7704) BglII (7558) NcoI (7543) RsrII (6895) PmlI (6627) BstEII (5707) BlpI (5560) HpaI (5033) NsiI (4913) RB T-DNA repeat PasI (1306) AclI (1900) BsiWI (2860) NheI (2976) BmtI (2980) EcoNI (3289) BsaI (3379) AgeI (3470) BstZ17I (3960) pPZP212 9015 bp
PmeI  (8863)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (8584)
1 site
G A A T T C C T T A A G
SacI  (8582)
1 site
G A G C T C C T C G A G
Eco53kI  (8580)
1 site
G A G C T C C T C G A G
KpnI  (8576)
1 site
G G T A C C C C A T G G
Acc65I  (8572)
1 site
G G T A C C C C A T G G
SmaI  (8570)
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  (8568)
1 site
C C C G G G G G G C C C
XmaI  (8568)
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.
BamHI  (8563)
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.
XbaI  (8557)
1 site
T C T A G A A G A T C T
SalI  (8551)
1 site
G T C G A C C A G C T G
PstI  (8549)
1 site
C T G C A G G A C G T C
SbfI  (8549)
1 site
C C T G C A G G G G A C G T C C
HindIII  (8533)
1 site
A A G C T T T T C G A A
BstXI  (8356)
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.
EcoRV  (7704)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
BglII  (7558)
1 site
A G A T C T T C T A G A
NcoI  (7543)
1 site
C C A T G G G G T A C C
RsrII  (6895)
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.
PmlI  (6627)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BstEII  (5707)
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.
BlpI  (5560)
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.
HpaI  (5033)
1 site
G T T A A C C A A T T G
NsiI  (4913)
1 site
A T G C A T T A C G T A
PasI  (1306)
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  (1900)
1 site
A A C G T T T T G C A A
BsiWI  (2860)
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.
NheI  (2976)
1 site
G C T A G C C G A T C G
BmtI  (2980)
1 site
G C T A G C C G A T C G
EcoNI  (3289)
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.
BsaI  (3379)
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.
AgeI  (3470)
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.
BstZ17I  (3960)
1 site
G T A T A C C A T A T G
pVS1 RepA
2245 .. 3318  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
pVS1 RepA
2245 .. 3318  =  1074 bp
357 amino acids  =  39.9 kDa
Product: replication protein from Pseudomonas
plasmid pVS1
NeoR/KanR
6749 .. 7546  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
6749 .. 7546  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
SmR
5080 .. 5871  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and
streptomycin
SmR
5080 .. 5871  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
confers resistance to spectinomycin and
streptomycin
CaMV 35S promoter (enhanced)
7606 .. 8284  =  679 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
7606 .. 8284  =  679 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
pVS1 StaA
1187 .. 1816  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
pVS1 StaA
1187 .. 1816  =  630 bp
209 amino acids  =  22.1 kDa
Product: stability protein from Pseudomonas
plasmid pVS1
ori
4248 .. 4836  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4248 .. 4836  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
lacZα
8370 .. 8603  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
8370 .. 8603  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
pVS1 oriV
3384 .. 3578  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
pVS1 oriV
3384 .. 3578  =  195 bp
origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)
CaMV poly(A) signal
6521 .. 6695  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
6521 .. 6695  =  175 bp
cauliflower mosaic virus polyadenylation signal
bom
3922 .. 4062  =  141 bp
basis of mobility region from pBR322
bom
3922 .. 4062  =  141 bp
basis of mobility region from pBR322
lac promoter
8647 .. 8677  =  31 bp
   Segment 3:  -10  
   8647 .. 8653  =  7 bp
promoter for the E. coli lac operon
lac promoter
8647 .. 8677  =  31 bp
   Segment 2:  
   8654 .. 8671  =  18 bp
promoter for the E. coli lac operon
lac promoter
8647 .. 8677  =  31 bp
   Segment 1:  -35  
   8672 .. 8677  =  6 bp
promoter for the E. coli lac operon
lac promoter
8647 .. 8677  =  31 bp
3 segments
promoter for the E. coli lac operon
LB T-DNA repeat
6399 .. 6423  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6399 .. 6423  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8878 .. 8902  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8878 .. 8902  =  25 bp
right border repeat from nopaline C58 T-DNA
lac operator
8623 .. 8639  =  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
8623 .. 8639  =  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
8533 .. 8589  =  57 bp
pUC18 multiple cloning site
MCS
8533 .. 8589  =  57 bp
pUC18 multiple cloning site
M13 fwd
8513 .. 8529  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
8513 .. 8529  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
8599 .. 8615  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
8599 .. 8615  =  17 bp
common sequencing primer, one of multiple similar
variants
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