pPZP101

Agrobacterium binary vector for plant transformation, with a chloramphenicol-resistance gene. The MCS is reversed in pPZP102.
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PmeI (6876) PvuI (6796) HindIII (6670) PstI - SbfI (6662) SalI (6652) XbaI (6646) BamHI (6640) SmaI (6637) KpnI - TspMI - XmaI (6635) Acc65I (6631) BanII - SacI (6629) Eco53kI (6627) EcoRI (6619) lac operator LB T-DNA repeat BclI * (6056) BspEI (5552) BpmI (5432) SspI (5240) HpaI (5033) NsiI (4913) BssSI - BssSĪ±I (4360) BstZ17I (3960) PluTI (3765) SfoI (3763) NarI (3762) KasI (3761) BbsI (56) BsaBI * (1299) BsiWI (2860) NheI (2976) BmtI (2980) BspDI * - ClaI * (3056) BseRI (3253) EcoNI (3289) BsaI (3379) AgeI (3470) pPZP101 7028 bp
PmeI  (6876)
1 site
G T T T A A A C C A A A T T T G
PvuI  (6796)
1 site
C G A T C G G C T A G C
HindIII  (6670)
1 site
A A G C T T T T C G A A
PstI  (6662)
1 site
C T G C A G G A C G T C
SbfI  (6662)
1 site
C C T G C A G G G G A C G T C C
SalI  (6652)
1 site
G T C G A C C A G C T G
XbaI  (6646)
1 site
T C T A G A A G A T C T
BamHI  (6640)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HFĀ® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
SmaI  (6637)
1 site
C C C G G G G G G C C C

SmaI can be used at 37Ā°C for brief incubations.
KpnI  (6635)
1 site
G G T A C C C C A T G G
TspMI  (6635)
1 site
C C C G G G G G G C C C
XmaI  (6635)
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  (6631)
1 site
G G T A C C C C A T G G
BanII  (6629)
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  (6629)
1 site
G A G C T C C T C G A G
Eco53kI  (6627)
1 site
G A G C T C C T C G A G
EcoRI  (6619)
1 site
G A A T T C C T T A A G
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 50-55Ā°C, but is 50% active at 37Ā°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 37Ā°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
BssSI  (4360)
1 site
C A C G A G G T G C T C
BssSĪ±I  (4360)
1 site
C A C G A G G T G C T C
BstZ17I  (3960)
1 site
G T A T A C C A T A T 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.
SfoI  (3763)
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.
KasI  (3761)
1 site
G G C G C C C C G C G G
BbsI  (56)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20Ā°C.
BsaBI  (1299)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BsiWI  (2860)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55Ā°C, but is 50% active at 37Ā°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.
BseRI  (3253)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37Ā°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
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 37Ā°C and 50Ā°C.
AgeI  (3470)
1 site
A C C G G T T G G C C A
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
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
lacZĪ±
6605 .. 6835  =  231 bp
76 amino acids  =  8.5 kDa
Product: LacZĪ± fragment of Ī²-galactosidase
lacZĪ±
6605 .. 6835  =  231 bp
76 amino acids  =  8.5 kDa
Product: LacZĪ± fragment of Ī²-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)
bom
3922 .. 4062  =  141 bp
basis of mobility region from pBR322
bom
3922 .. 4062  =  141 bp
basis of mobility region from pBR322
lac promoter
6531 .. 6561  =  31 bp
3 segments
   Segment 1:  -35  
   6531 .. 6536  =  6 bp
promoter for the E. coli lac operon
lac promoter
6531 .. 6561  =  31 bp
3 segments
   Segment 2:  
   6537 .. 6554  =  18 bp
promoter for the E. coli lac operon
lac promoter
6531 .. 6561  =  31 bp
3 segments
   Segment 3:  -10  
   6555 .. 6561  =  7 bp
promoter for the E. coli lac operon
lac promoter
6531 .. 6561  =  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
6891 .. 6915  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
6891 .. 6915  =  25 bp
right border repeat from nopaline C58 T-DNA
lac operator
6569 .. 6585  =  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-Ī²-D-thiogalactopyranoside (IPTG).
lac operator
6569 .. 6585  =  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-Ī²-D-thiogalactopyranoside (IPTG).
MCS
6619 .. 6675  =  57 bp
pUC18 multiple cloning site
MCS
6619 .. 6675  =  57 bp
pUC18 multiple cloning site
M13 rev
6593 .. 6609  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
6593 .. 6609  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
6679 .. 6695  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
6679 .. 6695  =  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:  1187 .. 1816  =  630 bp
ORF:  209 amino acids  =  22.1 kDa
ORF:  6605 .. 6835  =  231 bp
ORF:  76 amino acids  =  8.5 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:  653 .. 925  =  273 bp
ORF:  90 amino acids  =  9.9 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
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