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Following initial homologous recombination, enrichment of PrfA* (G145S) mutants was by performed by passaging merodiploid strains at 30C shaking in selective Listeria Synthetic Media (LSM) with 55mM glucose-6-phosphate without glutathione (Sigma-Aldrich G7879) and plated on p-chlorophenylalanine as a counter selectable marker

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In agreement with observations, the associated simulated trend in the water isotope 18 O is weak ( per decade)