Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010830 | pPZP200 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
The pPZP200 is a agrobacterium binary vector for plant transformation, with a spectinomycin-resistance gene.
- Vector Name:
- pPZP200
- Antibiotic Resistance:
- Streptomycin
- Length:
- 6742 bp
- Type:
- Plant Vectors
- Replication origin:
- ori
- Source/Author:
- Hajdukiewicz P, Svab Z, Maliga P.
- Copy Number:
- High copy number
- Growth Strain(s):
- JM108
- Growth Temperature:
- 37℃
pPZP200 vector Vector Map
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5. Store the plasmid at -20 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- Hajdukiewicz P, Svab Z, Maliga P. The small, versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol. 1994 Sep;25(6):989-94. doi: 10.1007/BF00014672. PMID: 7919218.
- Gupta SK, Vishwakarma NK, Malakar P, Vanspati P, Sharma NK, Chattopadhyay D. Development of an Agrobacterium-delivered codon-optimized CRISPR/Cas9 system for chickpea genome editing. Protoplasma. 2023 Sep;260(5):1437-1451. doi: 10.1007/s00709-023-01856-4. Epub 2023 May 3. PMID: 37131068.
pPZP200 vector Sequence
LOCUS 40924_35808 6742 bp DNA circular SYN 01-JAN-1980
DEFINITION Agrobacterium binary vector for plant transformation, with a
spectinomycin-resistance gene.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6742)
AUTHORS Hajdukiewicz P, Svab Z, Maliga P.
TITLE The small, versatile pPZP family of Agrobacterium binary vectors for
plant transformation.
JOURNAL Plant Mol. Biol. 1994;25:989-94.
PUBMED 7919218
REFERENCE 2 (bases 1 to 6742)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6742)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant Mol.
Biol."; date: "1994"; volume: "25"; pages: "989-94"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT The GenBank sequence was corrected by inserting a G at position
2249.
FEATURES Location/Qualifiers
source 1..6742
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1187..1813
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 2245..3315
/codon_start=1
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 3384..3578
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 3922..4062
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(4248..4836)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5083..5871)
/codon_start=1
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_feature 6399..6423
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
misc_feature 6492..6548
/label=MCS
/note="pUC18/19 multiple cloning site"
misc_feature 6605..6629
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"