pET-28a(+)::NL
(Plasmid #141289)

• Purpose: NLUC in pET-28 a (+) backbone for bacterial IPTG inducible expression (KmR)
• Depositing Lab: Andrew Millar
• Publication: Urquiza-Garcia et al Plant Methods. 2019 Jul 8;15:68. doi: 10.1186/s13007-019-0454-4. eCollection 2019.

Backbone

• Vector backbone: pET-28 a (+)
• Backbone manufacturer: EMD Biosciences
• Backbone size w/o insert (bp): 5369
• Total vector size (bp): 5796
• Vector type: Bacterial Expression, Luciferase

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Kanamycin, 25 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): BL21 DE3 Rossetta2 pLysS (Novagen, Merck, Darmstadt, Germany)
• Growth instructions: LB media with 50 µg/ml Kanamycin (Kn50) and 34 µg/ml Chloramphenicol (Cm34). Incubate the plate over-night at 37 °C. The chloramphenicol relates to the bacterial strain (E. coli BL21 DE3 Rossetta2 pLysS) and not the plasmid itself
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: NLUC
• Alt name: NanoLuc
• Alt name: NanoLUC
• Alt name: NanoLuciferase
• Species: Synthetic; Oplophorus gracilirostris
• Insert Size (bp): 513
• Mutation: Engineered for high stability (t1/2 = 11.5 days at 37 °C) and codon optimised for expression in mammalian cells.
• Promoter: promoter for bacteriophage T7 RNA polymerase

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TAATACGACTCACTATAGGG

Resource Information

• Supplemental Documents:
  • pet28ananoluc.gb
• A portion of this plasmid was derived from a plasmid made by: Plasmids pNL1.1 and pNL1.2 with NanoLUC sequence were kindly provided by Promega Corporation (UK, Delta House, Southampton Science Park, Southampton, SO16 7NS).

Terms and Licenses

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