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. 2024 Aug 28;13(9):2804–2819. doi: 10.1021/acssynbio.4c00186

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© 2024 The Authors. Published by American Chemical Society

Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).

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Construct design and protein schematics of the engineered auxin biosensors. The single-fusion (in trans) designs consist of a Venus-Aux/IAA17 fusion and free mScarlet-I expressed from the same cistron. The bicistronic equine rhinitis B virus (ERBV) 2A self-cleaving peptide is inserted into the cassettes. The auxin receptor, TIR or AFB2, is expressed separately from another construct. The dual-fusion (in cis) designs consist of a Venus-Aux/IAA17 fusion and TIR1- or AFB2-mScarlet-I fusion expressed from a single mRNA with ERBV-2A inserted between them, offering fewer steps of genetic manipulation and cell transformation.