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. 2021 Jan 20;217(3):iyab006. doi: 10.1093/genetics/iyab006

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© The Author(s) 2021. Published by Oxford University Press on behalf of Genetics Society of America.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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The F2A ribosome skip sequence functions efficiently in an eft-3p::TIR1::F2A::BFP::AID*::NLS transgene. (A) L3 larvae expressing eft-3p:: TIR1::F2A::BFP::AID*::NLS. A control animal expresses AID*-tagged BFP in the nuclei of vulval precursor cells (VPCs; white arrows). BFP expression is undetectable in animals grown on 1 mM K-NAA—a water-soluble, synthetic auxin—for 1 hour before imaging. Scale bars represent 15 µm (eft-3p). (B) Western blot detecting BFP::AID::NLS. Stain-free (Bio-Rad) analysis of total protein on the blot is provided as a loading control (left). Marker size (in kilodaltons) is provided. Anti-BFP blot showing background bands (marked with *) and a doublet consistent with the predicted size of BFP::AID*::NLS (black arrow) at approximately 34.5 kDa, and a smaller band below, likely a BFP degradation product.