p-mCherry-N1-slowChloC
(Plasmid #85462)

• Purpose: Historical plasmid. For inhibition of neuronal activity, #66709 is a light-gated chloride channel with improved anion selectivity (iChloC)
• Depositing Lab: Peter Hegemann
• Publication: Wietek et al Science. 2014 Mar 27.

Backbone

• Vector backbone: pEGFP-N1
• Backbone size w/o insert (bp): 4733
• Total vector size (bp): 5660
• Modifications to backbone: EGFP has been replaced by inserting the complete fusion construct (ChR2-mCherry) using XbaI and BamHI cloning sites
• 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: slowChloC
• Alt name: Channelrhodopsin-2
• Alt name: ChR2
• Species: Synthetic; Chlamydomonas reinhardtii
• Insert Size (bp): 927
• Mutation: E90R, T159C, D156N
• GenBank ID: EU714030.1
• Promoter: CMV (+enhancer)
• Tag / Fusion Protein:
  • mCherry (C terminal on insert)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • 53_slowChloC_mCherry.gb

Terms and Licenses

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