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Addgene

pcDNA3 Plk4(Sak) D154A (Nigg HR26)
(Plasmid #41164)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 41164 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pcDNA3.1/3x myc-A
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5500
  • Total vector size (bp) 8400
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Plk4 D154A
  • Alt name
    polo-like kinase 4
  • Alt name
    SAK D154A
  • Alt name
    STK18
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    2900
  • Mutation
    D154A
  • Entrez Gene
    PLK4 (a.k.a. MCCRP2, SAK, STK18)
  • Promoter CMV
  • Tags / Fusion Proteins
    • 3xMyc (N terminal on backbone)
    • PreScission Site (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer BGH-rev
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Human PLK4 cDNA amplified from expressed sequence tag AI561146 clone IMAGE:2210962 (RZPD, Berlin, Germany).
  • Article Citing this Plasmid

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Polymerase chain reaction was used to amplify full-length human PLK4 from the expressed sequence tag AI561146 clone IMAGE:2210962 (RZPD, Berlin, Germany) and to mutate codon 154 (GAT) to GCT, generating the catalytically inactive PLK4 (D154A) mutant. Both cDNAs were then subcloned into mammalian expression vector pcDNA3.1/3x myc-A providing an N-terminal 3xMyc tag. Both constructs were verified by sequencing.

Addgene NGS results found additional amino acids - ESRGPV - at the very C-terminus of the insert compared to the NCBI reference [AAX43264.1], but construct should function as described in the associated publication.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pcDNA3 Plk4(Sak) D154A (Nigg HR26) was a gift from Erich Nigg (Addgene plasmid # 41164 ; http://n2t.net/addgene:41164 ; RRID:Addgene_41164)
  • For your References section:

    The Polo kinase Plk4 functions in centriole duplication. Habedanck R, Stierhof YD, Wilkinson CJ, Nigg EA. Nat Cell Biol. 2005 Nov;7(11):1140-6. 10.1038/ncb1320 PubMed 16244668