pAAV-CaMKII-GAL4-iLight-VP16
(Plasmid #170274)

• Purpose: NLS-Gal4(1-65)-iLight(human codon optimized)-VP16 construct expression in mammalian cells. pAAV vector.
• Depositing Lab: Vladislav Verkhusha
• Publication: Kaberniuk et al Nat Commun. 2021 Jun 23;12(1):3859. doi: 10.1038/s41467-021-24212-7.

Backbone

• Vector backbone: pAAV-CW3SL-EGFP
• Backbone manufacturer: Addgene #61463
• Backbone size w/o insert (bp): 3736
• Vector type: AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: NLS-Gal4(1-65)-iLight(human codon optimized)-VP16
• Species: Bradyrhizobium sp. (strain ORS 278)
• Insert Size (bp): 2381
• Mutation: See Comments
• Promoter: CaMKII

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 5′ sequencing primer: pBR322ori-F
• 3′ sequencing primer: WPRE-R

Resource Information

• Supplemental Documents:
  • pAAV-CaMKII-GAL4-iLight-VP16.gb

Terms and Licenses

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

Depositor Comments

To design iLight, a full-length bacterial phytochrome IsPadC from Idiomarina sp. A28L was used as an original template. First, diguanylyl cyclase effector in IsPadC was removed, then following mutations were introduced: I68F, H80Q, A86T, R90S S242C, E274K, R295H I360V, and L464V. After that, iLight was human codon optimized for mammalian expression.