GFP-MYO10-BioID
(Plasmid #194858)

• Purpose: Express MYO10 tagged with eGFP (Nter) and BioID (Cter) in mammalian cells.
• Depositing Lab: Guillaume Jacquemet
• Publication: Popovic et al J Cell Sci. 2023 Mar 1;136(5):jcs260574. doi: 10.1242/jcs.260574. Epub 2023 Mar 2.

Backbone

• Vector backbone: EGFPC1
• 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: MYO10
• Alt name: myosin X
• Species: H. sapiens (human)
• Mutation: MyoX contains a Q680R variant compared to the NCBI reference [AAF37875.1]. The plasmid should function as described in the associated publication.
• Entrez Gene: MYO10 (a.k.a. MyoX)
• Promoter: CMV
• Tags / Fusion Proteins:
  • EGFP (N terminal on insert)
  • BioID (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: EGFP-C
• 3′ sequencing primer: SV40pA-R

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The original EGFPC1-hMyoX plasmid (Addgene plasmid # 47608) was created by Emanuel Strehler. The original BioID pcDNA3.1 MCS-BirA(R118G)-HA plasmid (Addgene plasmid # 36047) was created by Kyle Roux.

Terms and Licenses

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

Depositor Comments

The GFP-MYO10-BioID construct was generated as follows. Flanking XbaI sites were introduced into BioID by PCR (5’-ATTAGATCTAGAGGATCCAAGGACAACACCGTGCCCCTG, 5’-ATTAGATCTAGACTATGCGTAATCCGGTACATCGTAA, template plasmid: BioID pcDNA3.1 MCS-BirA(R118G)-HA) (Addgene plasmid # 36047) and the resulting amplicon was then inserted into a unique XbaI site in the EGFPC1-hMyoX plasmid (Addgene plasmid # 47608) resulting in an EGFP-MYO10-(stop codon)-BioID fusion gene. The stop codon between MYO10 and BioID was then replaced with a codon encoding valine (GTA) using a quick-change mutagenesis kit from Agilent and following the manufacturers' instructions.