pcDNA3.1+CMV-Ins-GLuc
(Plasmid #89929)

• Purpose: Expresses Insulin-GLuc from the CMV promoter. Gaussia Luc is inserted within the C-peptide and is co-secreted with insulin.
• Depositing Lab: Melanie Cobb
• Publication: Kalwat et al ACS Sens. 2016 Oct 28;1(10):1208-1212. Epub 2016 Oct 12.

Backbone

• Vector backbone: pcDNA3.1+
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5354
• Total vector size (bp): 6212
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: human insulin-Gaussia-Luciferase
• Alt name: hIns-GLuc
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 849
• Mutation: DNA sequence for Gaussia luciferase was humanized and two mutations (M43I/M110I) were made to enhance glow-like kinetics.
• Entrez Gene: INS (a.k.a. IDDM, IDDM1, IDDM2, ILPR, IRDN, MODY10, PNDM4)
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (unknown if destroyed)
• 3′ cloning site: XhoI (unknown if destroyed)
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: bGHrev

Resource Information

• A portion of this plasmid was derived from a plasmid made by: GeneArt Gene Synthesis (Life Technologies) was used to assemble human insulin with humanized Gaussia luciferase inserted into the C-peptide.

Terms and Licenses

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