pmCherry-3xNLS-OptoJNKi-lsm
(Plasmid #89742)

• Purpose: Expression of light-regulated JNK inhibitor, mCherry-tagged, localised to nucleus, 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: pmCherry-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4722
• Total vector size (bp): 5240
• Vector type: Mammalian Expression ; Fluorescent protein
• 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: NLS-OptoJNKi lit-state mutant
• Alt name: 3xNLS-AsLov2Jalpha (404-546)-I539E-JIP11
• Species: Synthetic; Avena Sativa and
• Insert Size (bp): 587
• Mutation: I539E mutation of AsLov2Jalpha
• GenBank ID: KY595936
• Promoter: CMV
• Tag / Fusion Protein:
  • mCherry (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsrGI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: DsRed_mCherry-fwd
• 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
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

For further information 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.