pF4Ag NanoLuc (ATG-42)
(Plasmid #137777)

• Purpose: Expresses NanoLuc in multi promoter expression vector (CMV andT7 promoters)
• Depositing Lab: Lance Encell
• Publication: Hall et al ACS Chem Biol. 2012 Nov 16;7(11):1848-57. doi: 10.1021/cb3002478. Epub 2012 Aug 30.

Backbone

• Vector backbone: pF4Ag
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 3827
• Total vector size (bp): 4340
• Modifications to backbone: none
• Vector type: Mammalian Expression, Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Depositor uses KRX (Promega) strain for protein production. For bacterial culture grow overnight in LB+100ug/ml ampicillin at 37C. To induce expression of NanoLuc dilute overnight culture 1:100 into LB supplemented with 0.1% Rhamnose and 0.15%Glucose+100ug/ml ampicillin and grow for 20 hours at 25C. NanoLuc expressed in this fashion will produce signal that is too bright for most luminometers and lysates require dilution of at least ~10,000 fold.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: NanoLuc (Nluc)
• Species: Oplophorus gracilirostris
• Insert Size (bp): 513
• Mutation: NanoLuc contains 16 amino acid changes compared to native Oplophorus gracilirostris
• Promoter: T7 and CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Sgf1 (unknown if destroyed)
• 3′ cloning site: Pme1 (not destroyed)
• 5′ sequencing primer: T7 promoter (TAA TAC GAC TCA CTA TAG GG)
• 3′ sequencing primer: T7 terminator (GCT AGT TAT TGC TCA GCG G)

Resource Information

• Supplemental Documents:
  • pF4Ag NanoLuc (ATG-42).gbk
• A portion of this plasmid was derived from a plasmid made by: Promega
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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