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The RT2 mix was added to the RT1 mixture (final volume: 25 L), and the reaction was carried out in a thermal cycler: 90 min at 37 C, followed by 5 min at 95 C, and held at 4 C

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Finally, a restraint-free production run was carried out for each simulation, with a time step of 2 fs in the NPT ensemble at 310 K and 1 bar using the Nose-Hoover thermostat and the semi-isotropic Parrinello-Rahman barostat 61 , respectively