Syn-ChR-tdTomato
(Plasmid #99843)

• Purpose: AAV expression of ChR2(H134R)-tdTomato under the Syn promoter
• Depositing Lab: Gordon Shepherd
• Publication: Shepherd Lab _ Syn-ChR-tdTomato plasmid (unpublished)

Backbone

• Vector backbone: pAAV
• Backbone manufacturer: Stratagene
• Backbone size w/o insert (bp): 4698
• Total vector size (bp): 7071
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: Optimal growth in Stbl3 at 30°C (use Carbenicillin).
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: ChR2(H134R)-tdTomato
• Alt name: Channelrhodopsin-2 H134R mutant
• Alt name: tandem dimer Tomato fluorescent protein
• Species: Chlamydomonas reinhardtii (modified)
• Insert Size (bp): 2373
• Mutation: ChR2(H134R), carrying a single point mutation at position H134, the native histidine residue is replaced by a more basic arginine.
• GenBank ID: EF474017.1 AY678269.1
• Promoter: human synapsin I promoter
• Tag / Fusion Protein:
  • tdTomato (C terminal on insert)

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: ggagccgtggcggcaccggtagtagcaGTGAGCAAGGGCG
• 3′ sequencing primer: gctttgtacggaattcTTACTTGTACAGCTCGT

Resource Information

• Supplemental Documents:
  • Syn_ChR2_tdTomato_sequencing_file_LLambot.gbk
• Addgene Notes:
  • Addgene Diagnostic Digest
• A portion of this plasmid was derived from a plasmid made by: Edward Boyden (Massachusetts Institute of Technology, Cambridge, MA), Scott Sternson (Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia).

Terms and Licenses

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

Depositor Comments

Backbone vector:
Stratagene

ChR2(H134R):
Edward Boyden
Boyden et al Nat Neurosci. 2005 Sep . 8(9):1263-8.

tdTomato:
Scott Sternson
Atasoy et al J Neurosci. 2008 Jul 9. 28(28):7025-30.