pPZP122

Agrobacterium binary vector for plant transformation, with chloramphenicol- and gentamycin-resistance genes. The MCS is reversed in pPZP121.
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PmeI (8475) lac operator EcoRI (8196) BanII - SacI (8194) Eco53kI (8192) KpnI (8188) Acc65I (8184) SmaI (8182) TspMI - XmaI (8180) BamHI (8175) XbaI (8169) SalI (8163) PstI - SbfI (8161) HindIII (8145) BstXI (7968) NcoI (7155) PmlI (6522) BclI * (6056) BspEI (5552) BpmI (5432) SspI (5240) HpaI (5033) NsiI (4913) RB T-DNA repeat BsiWI (2860) NheI (2976) BmtI (2980) BspDI * - ClaI * (3056) EcoNI (3289) BsaI (3379) AgeI (3470) KasI (3761) NarI (3762) SfoI (3763) PluTI (3765) BstZ17I (3960) pPZP122 8627 bp
PmeI  (8475)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (8196)
1 site
G A A T T C C T T A A G
BanII  (8194)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (8194)
1 site
G A G C T C C T C G A G
Eco53kI  (8192)
1 site
G A G C T C C T C G A G
KpnI  (8188)
1 site
G G T A C C C C A T G G
Acc65I  (8184)
1 site
G G T A C C C C A T G G
SmaI  (8182)
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  (8180)
1 site
C C C G G G G G G C C C
XmaI  (8180)
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  (8175)
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  (8169)
1 site
T C T A G A A G A T C T
SalI  (8163)
1 site
G T C G A C C A G C T G
PstI  (8161)
1 site
C T G C A G G A C G T C
SbfI  (8161)
1 site
C C T G C A G G G G A C G T C C
HindIII  (8145)
1 site
A A G C T T T T C G A A
BstXI  (7968)
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.
NcoI  (7155)
1 site
C C A T G G G G T A C C
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
BpmI  (5432)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
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
BspDI  (3056)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3056)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
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
KasI  (3761)
1 site
G G C G C C C C G C G G
NarI  (3762)
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.
SfoI  (3763)
1 site
G G C G C C C C G C G G
PluTI  (3765)
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.
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
CaMV 35S promoter (enhanced)
7218 .. 7896  =  679 bp
cauliflower mosaic virus 35S promoter with a duplicated enhancer region
CaMV 35S promoter (enhanced)
7218 .. 7896  =  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
GmR
6625 .. 7158  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
6625 .. 7158  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
lacZα
7982 .. 8215  =  234 bp
77 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
7982 .. 8215  =  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
8259 .. 8289  =  31 bp
3 segments
   Segment 3:  -10  
   8259 .. 8265  =  7 bp
promoter for the E. coli lac operon
lac promoter
8259 .. 8289  =  31 bp
3 segments
   Segment 2:  
   8266 .. 8283  =  18 bp
promoter for the E. coli lac operon
lac promoter
8259 .. 8289  =  31 bp
3 segments
   Segment 1:  -35  
   8284 .. 8289  =  6 bp
promoter for the E. coli lac operon
lac promoter
8259 .. 8289  =  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
8490 .. 8514  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
8490 .. 8514  =  25 bp
right border repeat from nopaline C58 T-DNA
lac operator
8235 .. 8251  =  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
8235 .. 8251  =  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
8145 .. 8201  =  57 bp
pUC18 multiple cloning site
MCS
8145 .. 8201  =  57 bp
pUC18 multiple cloning site
M13 fwd
8125 .. 8141  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
8125 .. 8141  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
8211 .. 8227  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
8211 .. 8227  =  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:  6031 .. 6372  =  342 bp
ORF:  113 amino acids  =  12.1 kDa
ORF:  6763 .. 7095  =  333 bp
ORF:  110 amino acids  =  11.9 kDa
ORF:  1187 .. 1816  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  5919 .. 6161  =  243 bp
ORF:  80 amino acids  =  9.1 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:  7323 .. 7640  =  318 bp
ORF:  105 amino acids  =  11.6 kDa
ORF:  7650 .. 7988  =  339 bp
ORF:  112 amino acids  =  12.4 kDa
ORF:  653 .. 925  =  273 bp
ORF:  90 amino acids  =  9.9 kDa
ORF:  7982 .. 8215  =  234 bp
ORF:  77 amino acids  =  8.6 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:  5107 .. 5907  =  801 bp
ORF:  266 amino acids  =  31.0 kDa
ORF:  6625 .. 7158  =  534 bp
ORF:  177 amino acids  =  19.4 kDa
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