pWPI/hPLK2K111M/Neo
(Plasmid #35386)

• Depositing Lab: Anurag Tandon
• Publication: Visanji et al J Biol Chem. 2011 Oct 14;286(41):35863-73. Epub 2011 Aug 17.

Backbone

• Vector backbone: pWPI/Neomycin
• Backbone manufacturer: Didier Trono
• Backbone size w/o insert (bp): 11437
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: hPLK2 K111M
• Alt name: Snk/Plk2
• Species: H. sapiens (human)
• Mutation: K111M
• Entrez Gene: PLK2 (a.k.a. SNK, hPlk2, hSNK)
• Promoter: EF1-alpha
• Tags / Fusion Proteins:
  • V5 epitope and Polyhistidine tag (C terminal on insert)
  • EMCV IRES-Neomycin cassette (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: EF-1a-F
• 3′ sequencing primer: EGFP-N

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The backbone of this bicistronic lentiviral vector was originally created by Dr. Didier Trono lab (pWPI, Plasmid # 12254), and then modified by Robert Strome using Neomycin to replace EGFP and adding several multiple clone sites. The MCS includes Swa I, Nde I, BamH I, Spe I, Xma I and Sma I (see sequence). The hPlk2WT cDNA was inserted with BamH I site, and can be replaced by any other gene with this site. This vector allows the simultaneous expression of hPlk2K111M and Neomycin to facilitate establishing stable mammalian transgene cell lines.
Please note that the full sequence for this plasmid is approximated and not fully verified. For further cloning strategies, please contact with Dr. Anurag Tandon.