pIZ-EGFP-MYO10-HMM
(Plasmid #87257)

• Purpose: Expression plasmid to perform NanoSPD assays in insect cells (generates EGFP-tagged bait).
• Depositing Lab: Thomas Friedman
• Publication: Bird et al Mol Biol Cell. 2016 Dec 8. pii: mbc.E16-08-0583.

Backbone

• Vector backbone: pIZ/V5-His
• Backbone manufacturer: Thermo Scientific
• Backbone size w/o insert (bp): 2900
• Total vector size (bp): 6431
• Modifications to backbone: When cloning into the MCS to generate a EGFP-Myo10 fusion protein, a stop codon must be included to prevent translation into the C-terminal V5/His tag.
• Vector type: Insect Expression
• Selectable markers: Zeocin

Growth in Bacteria

• Bacterial Resistance(s): Bleocin (Zeocin), 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Zeocin is sensitive to NaCl in standard LB and LB-Agar formulations. Use modified recipe, into 1L ddH20 add: 10g tryptone, 5g yeast extract, 5g NaCl.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: MYO10
• Alt name: Unconventional myosin 10
• Species: M. musculus (mouse)
• Insert Size (bp): 2823
• Mutation: Myo10 is truncated to contain aa.1-941
• GenBank ID: NM_019472.2
• Entrez Gene: Myo10 (a.k.a. AW048724, D15Ertd600, D15Ertd600e, mKIAA0799, myos, myosin-X)
• Promoter: OpIE2

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: OpIE2_fwd_primer

Resource Information

• Supplemental Documents:
  • pIZ-EGFP-Myo10-HMM.gbk

Terms and Licenses

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