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

• Depositing Lab: Erich Nigg
• Publication: Habedanck et al Nat Cell Biol. 2005 Nov;7(11):1140-6.

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

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:
  • 1 Reference

Terms and Licenses

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

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.