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Addgene

HA.PKD.K/W
(Plasmid #10809)

Full plasmid sequence is not available for this item.

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 10809 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pcDNA3
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5446
  • 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
    PKD
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    2800
  • Mutation
    Kinase dead mutant: K612W (Lys 612 to Trp) (bp1834-1836 AAA -> TGG); Also Q66R, F139S and G877W
  • Entrez Gene
    PRKD1 (a.k.a. CHDED, PKC-MU, PKCM, PKD, PRKCM)
  • Promoter CMV
  • Tags / Fusion Proteins
    • HA (N terminal on insert)
    • Myc (inserted between amino acids 33&34 of PKD)

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

Terms and Licenses

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

Depositor Comments

The HA tag on this plasmid contains a point mutation that renders it unrecognizable by the 3F10 HA antibody. It will work fine with the 12CA5 HA antibody.

The PKD insert in this plasmid contains a Myc tag inserted between amino acids 33&34, Q66R, F139S and G877W mutations when compared to GenBank reference sequence NP_002733.1. Although the effect of these mutations is unknown, this is the exact plasmid used in the associated publication and it should function as described.

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:

    HA.PKD.K/W was a gift from Alex Toker (Addgene plasmid # 10809 ; http://n2t.net/addgene:10809 ; RRID:Addgene_10809)
  • For your References section:

    Tyrosine phosphorylation of protein kinase D in the pleckstrin homology domain leads to activation. Storz P, Doppler H, Johannes FJ, Toker A. J Biol Chem. 2003 May 16. 278(20):17969-76. 10.1074/jbc.M213224200 PubMed 12637538