pTol1-UAS:CheRiff-tdTomato
(Plasmid #124234)

• Purpose: Zebrafish Tol1 construct for UAS-mediated expression of the optogenetic opsin CheRiff fused to the red fluorescent protein tdTomato
• Depositing Lab: Isaac Bianco
• Publication: Antinucci et al Elife. 2020 Mar 27;9. pii: 54937. doi: 10.7554/eLife.54937.

Backbone

• Vector backbone: pT1UciMP
• Backbone manufacturer: from Harold Burgess lab (Addgene Plasmid #62215)
• Backbone size w/o insert (bp): 4548
• Total vector size (bp): 6944
• Modifications to backbone: NcoI linearization before insert cloning
• Vector type: Zebrafish expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: CheRiff-tdTomato
• Species: Scherffelia dubia
• Insert Size (bp): 2394
• GenBank ID:
• Promoter: 14xUAS-E1b
• Tag / Fusion Protein:
  • tdTomato (C terminal on insert)

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: TCA GTC GTC CGT TTG TTC CG
• 3′ sequencing primer: GAG CAG CAA AGC CAT GTA GC

Resource Information

• Supplemental Documents:
  • pTol1-UAS_CheRiff-tdTomato.gb
• A portion of this plasmid was derived from a plasmid made by: DRH313: FCK-CheRiff-eGFP (Addgene Plasmid #51693). All-optical electrophysiology in mammalian neurons using engineered microbial rhodopsins. Hochbaum DR, Zhao Y, Farhi SL, Klapoetke N, Werley CA, Kapoor V, Zou P, Kralj JM, Maclaurin D, Smedemark-Margulies N, Saulnier JL, Boulting GL, Straub C, Cho YK, Melkonian M, Wong GK, Harrison DJ, Murthy VN, Sabatini BL, Boyden ES, Campbell RE, Cohen AE. Nat Methods. 2014 Aug;11(8):825-33. doi: 10.1038/nmeth.3000. Epub 2014

Terms and Licenses

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

Depositor Comments

Zebrafish Tol1 construct for UAS-mediated expression of the optogenetic opsin CoChR fused to the red fluorescent protein tdTomato. In addition to the opsin gene, it contains tol1 arms, 14xUAS-E1b promoter, ubiquitin C intron, and ocean pout antifreeze protein terminator. Co-inject with Tol1 (not Tol2) transposase mRNA.