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) 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

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
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
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
3 segments
   Segment 3:  -10  
   8647 .. 8653  =  7 bp
promoter for the E. coli lac operon
lac promoter
8647 .. 8677  =  31 bp
3 segments
   Segment 2:  
   8654 .. 8671  =  18 bp
promoter for the E. coli lac operon
lac promoter
8647 .. 8677  =  31 bp
3 segments
   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
ORF:  202 .. 888  =  687 bp
ORF:  228 amino acids  =  25.1 kDa
ORF:  2143 .. 3318  =  1176 bp
ORF:  391 amino acids  =  43.9 kDa
ORF:  5449 .. 5832  =  384 bp
ORF:  127 amino acids  =  14.6 kDa
ORF:  6136 .. 6477  =  342 bp
ORF:  113 amino acids  =  12.1 kDa
ORF:  1187 .. 1816  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  5294 .. 5572  =  279 bp
ORF:  92 amino acids  =  9.4 kDa
ORF:  5582 .. 5839  =  258 bp
ORF:  85 amino acids  =  9.1 kDa
ORF:  6980 .. 7234  =  255 bp
ORF:  84 amino acids  =  9.7 kDa
ORF:  6024 .. 6266  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  925 .. 1239  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2113 .. 2685  =  573 bp
ORF:  190 amino acids  =  20.6 kDa
ORF:  5080 .. 6012  =  933 bp
ORF:  310 amino acids  =  34.5 kDa
ORF:  6985 .. 7371  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  7711 .. 8028  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
ORF:  8038 .. 8376  =  339 bp
ORF:  112 amino acids  =  12.4 kDa
ORF:  234 .. 584  =  351 bp
ORF:  116 amino acids  =  12.6 kDa
ORF:  2091 .. 2348  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  8370 .. 8603  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  653 .. 925  =  273 bp
ORF:  90 amino acids  =  9.9 kDa
ORF:  6749 .. 7546  =  798 bp
ORF:  265 amino acids  =  29.1 kDa
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