pET28a Gx-NL3
(Plasmid #169108)

• Purpose: Expresses protein G fused to 3 copies of NanoLuc. The protein G domain contains an amber stop-codon at position 24 for incorporation of the unnatural amino acid para-benzoylphenylalanine.
• Depositing Lab: Maarten Merkx
• Publication: Wouters et al Bioconjug Chem. 2020 Mar 18;31(3):656-662. doi: 10.1021/acs.bioconjchem.9b00804. Epub 2020 Jan 22.

Backbone

• Vector backbone: pET28a
• Backbone size w/o insert (bp): 5369
• Total vector size (bp): 7087
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Bl21(DE3)
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: protein G - NanoLuc - NanoLuc - NanoLuc
• Species: Synthetic
• Insert Size (bp): 1854
• Mutation: Ala 24 stop
• Promoter: T7
• Tags / Fusion Proteins:
  • strep-tag (N terminal on insert)
  • his-tag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (unknown if destroyed)
• 3′ cloning site: XhoI (unknown if destroyed)
• 5′ sequencing primer: T7

Resource Information

• Supplemental Documents:
  • pET28a_pGx-NL3.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • NanoLuc Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry