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Scheme of the evolutionary experiment setup with yvcK::Himar1 in BHI supplemented with 2 mM t-CIN as described in Materials and Methods. Isolates which recovered t-CIN tolerance were identified in all six independent lineages with t-CIN supplementation after three rounds of subculture, while no recovery of t-CIN tolerance was observed in control cultures without t-CIN. The molecular structure of t-CIN is also shown.
