pLuc-OptoJNKi-lsm
(Plasmid #89746)

• Purpose: Expression of light-regulated JNK inhibitor, firefly luciferase-fused, lit-state mutant photosensor (AsLov2Ja.I539E), inhibitor JIP11
• Depositing Lab: Michael Courtney
• Publication: Melero-Fernandez de Mera et al Nat Commun. 2017 May 12;8:15017. doi: 10.1038/ncomms15017.

Backbone

• Vector backbone: pLuc-C1
• Backbone manufacturer: Derived by our lab using pExFP-C1 and pGL3
• Backbone size w/o insert (bp): 5673
• Total vector size (bp): 6119
• Modifications to backbone: Firefly Luciferase in place of FP
• Vector type: Mammalian Expression, Luciferase
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: OptoJNKi lit-state mutant
• Alt name: AsLov2Jalpha (404-546)-I539E-JIP11
• Species: Synthetic; Avena Sativa and
• Insert Size (bp): 478
• Mutation: I539E mutation of AsLov2Jalpha
• GenBank ID: KY595942
• Promoter: CMV
• Tag / Fusion Protein:
  • Firefly luciferase (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: luc2131+
• 3′ sequencing primer: fastbac reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: We designed and assembled the insert from de novo synthesised sequences

Terms and Licenses

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

Depositor Comments

Please see
Melero-Fernandez de Mera RM*, Li LL*, Popinigis A, Cisek K, Tuittila M, Yadav L, Serva A, Courtney MJ (2017) A simple optogenetic MAPK inhibitor design reveals resonance between transcription-regulating circuitry and temporally-encoded inputs. Nat. Commun. 8, 15017 doi: 10.1038/ncomms15017 in press.