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pcDNA3 PBD HK-AA (Nigg AH13)
(Plasmid #41161)

Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 41161 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pcDNA3.1/3x myc-C
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5500
  • Total vector size (bp) 6331
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    PBD HK-AA
  • Alt name
    Plk1-PBD HK-AA
  • Alt name
    Plk1 Polo Box Domain
  • Alt name
    Polo-like kinase 1 Polo Box Domain
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    831
  • Mutation
    Contains Plk1-PBD only (aa 326–603); HK-AA (H538A, K540A)
  • GenBank ID
    NM_005030.3 X73458.1
  • Entrez Gene
    PLK1 (a.k.a. PLK, STPK13)
  • 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 EcoRI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer BGH-rev
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Addgene plasmid #41162: Plk1-PBDWT (aa 326–603) was cloned in-frame by PCR into a pcDNA3.1 vector encoding an amino-terminal 3xmyc-tag (Invitrogen, Carlsbad, CA) using primers 5'-CCG GAA TTC CAG ATC TTC GAT TGC TCC CAG CAG CCT GG-3' and
5'-CCG CTC GAG TTA GGA GGC CTT GAG ACG GTT GC-3'.

The Plk1-PBDAA mutant (H538A, K540A) was made by site-directed mutagenesis using the following primers: 5'-CAA TTC TCC GGA GCC CCG CCT ATC TGT CCC-3' and 5'-GGG ACA GAT AGG CGG GGC TCC GGA GAA TTG-3'.

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 PBD HK-AA (Nigg AH13) was a gift from Erich Nigg (Addgene plasmid # 41161 ; http://n2t.net/addgene:41161 ; RRID:Addgene_41161)
  • For your References section:

    Different Plk1 functions show distinct dependencies on Polo-Box domain-mediated targeting. Hanisch A, Wehner A, Nigg EA, Sillje HH. Mol Biol Cell. 2006 Jan;17(1):448-59. Epub 2005 Nov 2. 10.1091/mbc.e05-08-0801 PubMed 16267267