pPZP111

Agrobacterium binary vector for plant transformation, with chloramphenicol- and kanamycin-resistance genes. The MCS is reversed in pPZP112.
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PmeI (8758) HindIII (8545) PstI - SbfI (8537) SalI (8527) XbaI (8521) BamHI (8515) SmaI (8512) KpnI - TspMI - XmaI (8510) Acc65I (8506) SacI (8504) Eco53kI (8502) EcoRI (8494) lac operator BstXI (8251) EcoRV (7599) BglII (7453) NcoI (7438) BssHII (6908) RsrII (6790) PmlI (6522) BclI * (6056) BspEI (5552) SspI (5240) HpaI (5033) NsiI (4913) BsiWI (2860) NheI (2976) BmtI (2980) EcoNI (3289) BsaI (3379) AgeI (3470) BstZ17I (3960) pPZP111 8910 bp
PmeI  (8758)
1 site
G T T T A A A C C A A A T T T G
HindIII  (8545)
1 site
A A G C T T T T C G A A
PstI  (8537)
1 site
C T G C A G G A C G T C
SbfI  (8537)
1 site
C C T G C A G G G G A C G T C C
SalI  (8527)
1 site
G T C G A C C A G C T G
XbaI  (8521)
1 site
T C T A G A A G A T C T
BamHI  (8515)
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  (8512)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KpnI  (8510)
1 site
G G T A C C C C A T G G
TspMI  (8510)
1 site
C C C G G G G G G C C C
XmaI  (8510)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
Acc65I  (8506)
1 site
G G T A C C C C A T G G
SacI  (8504)
1 site
G A G C T C C T C G A G
Eco53kI  (8502)
1 site
G A G C T C C T C G A G
EcoRI  (8494)
1 site
G A A T T C C T T A A G
BstXI  (8251)
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  (7599)
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  (7453)
1 site
A G A T C T T C T A G A
NcoI  (7438)
1 site
C C A T G G G G T A C C
BssHII  (6908)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
RsrII  (6790)
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  (6522)
1 site
C A C G T G G T G C A C
BclI  (6056)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BspEI  (5552)
1 site
T C C G G A A G G C C T
SspI  (5240)
1 site
A A T A T T T T A T A A
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
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
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
6644 .. 7441  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
6644 .. 7441  =  798 bp
265 amino acids  =  29.1 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
CaMV 35S promoter (enhanced)
7501 .. 8179  =  679 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CaMV 35S promoter (enhanced)
7501 .. 8179  =  679 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CmR
5107 .. 5766  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
5107 .. 5766  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
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
8480 .. 8713  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
8480 .. 8713  =  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
6416 .. 6590  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
6416 .. 6590  =  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
8406 .. 8436  =  31 bp
3 segments
   Segment 1:  -35  
   8406 .. 8411  =  6 bp
promoter for the E. coli lac operon
lac promoter
8406 .. 8436  =  31 bp
3 segments
   Segment 2:  
   8412 .. 8429  =  18 bp
promoter for the E. coli lac operon
lac promoter
8406 .. 8436  =  31 bp
3 segments
   Segment 3:  -10  
   8430 .. 8436  =  7 bp
promoter for the E. coli lac operon
lac promoter
8406 .. 8436  =  31 bp
3 segments
promoter for the E. coli lac operon
LB T-DNA repeat
6294 .. 6318  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
6294 .. 6318  =  25 bp
left border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8773 .. 8797  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8773 .. 8797  =  25 bp
right border repeat from nopaline C58 T-DNA
lac operator
8444 .. 8460  =  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).