Dynactin-4/p62-3XHA
(Plasmid #31058)

• Depositing Lab: Vann Bennett
• Publication: Ayalon et al J Biol Chem. 2011 Mar 4. 286(9):7370-8.

Backbone

• Vector backbone: Modified pEGFP-N1 (see note)
• Backbone size w/o insert (bp): 4000
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Dynactin-4
• Alt name: p62
• Alt name: dynactin subunit 4 isoform c
• Species: H. sapiens (human)
• Entrez Gene: DCTN4 (a.k.a. DYN4, P62)
• Tag / Fusion Protein:
  • 3XHA (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: CMV-Fwd

Terms and Licenses

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

Depositor Comments

For the preparation of HA-dynactin-4, GFP was first cut out of pEGFP-N1 vector between HindIII and NotI, and in its place, we introduced a triple HA sequence that was synthesized by PCR and ligated into the vector. To a recombinant ORF sequence of dynactin-4, we added by PCR a 5′ XhoI site, and a HindIII site on its 3′ end. The dynactin-4 construct was then cloned into the modified 3XHA-containing vector.