p-mCherry-N1-iChloC_CA
(Plasmid #98171)

• Purpose: slow cycling (open for minutes) step-function artificial anion conducting channelrhodopsin (aACR). High light sensitivity. Activation with blue to green light, inactivation max with 605 nm (accelarates closure to ms). Codon optimized for mammalian expression.
• Depositing Lab: Peter Hegemann
• Publication: Wietek et al Sci Rep. 2017 Nov 2;7(1):14957. doi: 10.1038/s41598-017-14330-y.

Backbone

• Vector backbone: pEGFP-N1
• Backbone size w/o insert (bp): 4733
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Synthetic construct iChloC_C128A gene
• Alt name: iChloC
• Alt name: CrChR2
• Species: Synthetic
• Insert Size (bp): 927
• Mutation: E83Q, E90R, E101S, C128A, T159C, D156N
• Promoter: CMV (+enhancer)
• Tag / Fusion Protein:
  • mCherry (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: EGFP-N1-F: GAGGTCTATATAAGCAGAGC or CMV-F: GCAAATGGGCGGTAGGCGT
• 3′ sequencing primer: EGFP-C1-R: AACCATTATAAGCTGCAATAAAC

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Modifications to backbone: EGFP has been replaced by inserting the complete fusion construct (ChR2-mCherry) using XbaI and BamHI cloning sites.