BMR1_02g00615-bio-His
(Plasmid #108029)

• Purpose: Expresses enzymatically monobiotinylated full-length BMR1_02g00615 ectodomain with no N-linked glycans upon cotransfection with BirA in mammalian cells. Contains a C-terminal rat Cd4 d3+4 tag.
• Depositing Lab: Gavin Wright
• Publication: Elton et al Int J Parasitol. 2019 Feb;49(2):115-125. doi: 10.1016/j.ijpara.2018.10.003. Epub 2018 Oct 24.

Backbone

• Vector backbone: pTT3
• Backbone manufacturer: Yves Durocher
• Backbone size w/o insert (bp): 6670
• Modifications to backbone: Addition of CD4, biotinylation sequence, 6xhis tags
• Vector type: Mammalian Expression

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: BMR1_02g00615
• Species: B. microti
• Insert Size (bp): 477
• Mutation: all potential N-linked glycosylation sites (N-X-S/T, where X is not proline) were systematically removed by substituting alanine for serine/threonine at these sites.
• Promoter: CMV
• Tag / Fusion Protein:
  • ratCD4d3+4,biotinylation sequence, 6xHis (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: AscI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: EBVori-F (5'-CCACTCAGTGTTGGCAAATG-3')

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Made by gene synthesis. pTT3 was obtained from Yves Durocher, National Research Council of Canada

Terms and Licenses

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