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pPZP211

Agrobacterium binary vector for plant transformation, with spectinomycin- and kanamycin-resistance genes. The MCS is reversed in pPZP212.

 
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pPZP211.dna
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PmeI (8863) HindIII (8650) PstI - SbfI (8642) SalI (8632) XbaI (8626) BamHI (8620) SmaI (8617) KpnI - TspMI - XmaI (8615) Acc65I (8611) SacI (8609) Eco53kI (8607) EcoRI (8599) 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) pPZP211 9015 bp
PmeI  (8863)
1 site
G T T T A A A C C A A A T T T G
HindIII  (8650)
1 site
A A G C T T T T C G A A
PstI  (8642)
1 site
C T G C A G G A C G T C
SbfI  (8642)
1 site
C C T G C A G G G G A C G T C C
SalI  (8632)
1 site
G T C G A C C A G C T G
XbaI  (8626)
1 site
T C T A G A A G A T C T
BamHI  (8620)
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  (8617)
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  (8615)
1 site
G G T A C C C C A T G G
TspMI  (8615)
1 site
C C C G G G G G G C C C
XmaI  (8615)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
Acc65I  (8611)
1 site
G G T A C C C C A T G G
SacI  (8609)
1 site
G A G C T C C T C G A G
Eco53kI  (8607)
1 site
G A G C T C C T C G A G
EcoRI  (8599)
1 site
G A A T T C C T T A A G
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

PmlI gradually loses activity when stored at -20°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