Plasmid 35386: pWPI/hPLK2K111M/Neo

• Gene/insert name: hPLK2 K111M
• Alt name: Snk/Plk2
• Species: H. sapiens (human)
• Entrez Gene: PLK2 (SNK, hPlk2, hSNK)
  
• Fusion protein or tag: V5 epitope and Polyhistidine tag
• Terminal: C terminal on insert
• Fusion protein or tag: EMCV IRES-Neomycin cassette
• Terminal: C terminal on backbone
• Mutation: K111M
• Vector backbone: pWPI/Neomycin
  (Search Vector Database)
• Backbone manufacturer: Didier Trono
• Vector type: Mammalian Expression, Lentiviral
• Backbone size w/o insert (bp): 11437
• Promoter: EF1-alpha
• Cloning site 5': BamHI
• Site destroyed during cloning: No
• Cloning site 3': BamHI
• Site destroyed during cloning: No
• 5' sequencing primer: EF-1a-F
• 3' sequencing primer: EGFP-N
• Bacterial resistance(s) Ampicillin
• Growth strain(s) Stbl3
• Growth temperature (℃): 37
• High or low copy: High Copy
• Selectable markers: Neomycin
• Sequence: View sequences (3)
• Principal Investigator: Anurag Tandon
• Terms and Licenses: MTA

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.

Article: Effect of Ser-129 phosphorylation on interaction of alpha-synuclein with synaptic and cellular membranes. Visanji et al (J Biol Chem. 2011 Oct 14;286(41):35863-73. Epub 2011 Aug 17. PubMed)