pPZP200

Agrobacterium binary vector for plant transformation, with a spectinomycin-resistance gene.
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RB T-DNA repeat PmeI (6590) HindIII (6543) PstI - SbfI (6535) SalI (6525) XbaI (6519) BamHI (6513) SmaI (6510) KpnI - TspMI - XmaI (6508) Acc65I (6504) BanII - SacI (6502) Eco53kI (6500) EcoRI (6492) BstEII (5707) BpmI (5622) PvuI (5493) BssHII (5144) HpaI (5033) NsiI (4913) BssSI - BssSαI (4360) BstZ17I (3960) PluTI (3765) SfoI (3763) NarI (3762) KasI (3761) AgeI (3470) BsaI (3379) ScaI (3) BbsI (56) BsaBI * (1299) PasI (1306) AclI (1900) BsiWI (2860) NheI (2976) BmtI (2980) BspDI * - ClaI * (3056) FspI (3193) BseRI (3253) EcoNI (3289) pPZP200 6742 bp
PmeI  (6590)
1 site
G T T T A A A C C A A A T T T G
HindIII  (6543)
1 site
A A G C T T T T C G A A
PstI  (6535)
1 site
C T G C A G G A C G T C
SbfI  (6535)
1 site
C C T G C A G G G G A C G T C C
SalI  (6525)
1 site
G T C G A C C A G C T G
XbaI  (6519)
1 site
T C T A G A A G A T C T
BamHI  (6513)
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  (6510)
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  (6508)
1 site
G G T A C C C C A T G G
TspMI  (6508)
1 site
C C C G G G G G G C C C
XmaI  (6508)
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  (6504)
1 site
G G T A C C C C A T G G
BanII  (6502)
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  (6502)
1 site
G A G C T C C T C G A G
Eco53kI  (6500)
1 site
G A G C T C C T C G A G
EcoRI  (6492)
1 site
G A A T T C C T T A A G
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.
BpmI  (5622)
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.
PvuI  (5493)
1 site
C G A T C G G C T A G C
BssHII  (5144)
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.
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
BssSαI  (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
AgeI  (3470)
1 site
A C C G G T T G G C C A
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.
ScaI  (3)
1 site
A G T A C T T C A T G A
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 -20°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.
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
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.
FspI  (3193)
1 site
T G C G C A A C G C G T
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 37°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.
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
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
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
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
MCS
6492 .. 6548  =  57 bp
pUC18 multiple cloning site
MCS
6492 .. 6548  =  57 bp
pUC18 multiple cloning site
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
6605 .. 6629  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
6605 .. 6629  =  25 bp
right border repeat from nopaline C58 T-DNA
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:  6024 .. 6266  =  243 bp
ORF:  80 amino acids  =  9.1 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:  5080 .. 6012  =  933 bp
ORF:  310 amino acids  =  34.5 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
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