With automated vehicles (AVs), reversible lanes could be a sustainable transportation solution once there is vehicle-to-infrastructure connectivity informingAVs about the lane configuration changes. This paper introduced the reversible lane network design problem (RL-NDP), formulated in mixed-integer non-linear mathematical programming-both the traffic assignment and the reversible lane decisions were embedded. The model was applied on an hourly basis in the case study of the city of Delft, the Netherlands. Reversible lanes are examined under no traffic equilibrium (former paths are maintained); user-equilibrium (UE) assignment (AVs decide their own paths); and system-optimum (SO) traffic assignment (AVs are forced to follow SO paths). We found out that reversible lanes reduce congested roads, total travel times, and delays up to 36%, 9%, and 22%, respectively. The SO scenario was revealed to be beneficial in reducing the total travel time and congested roads in peak hours, whereas UE is equally optimal in the remaining hours. A dual-scenario mixing SO and UE throughout the day reduced congested roads, total travel times, and delay up to 40%, 8%, and 19%, respectively, yet increased 1% in travel distance. The spatial analysis suggested a substantial lane variability in the suburbs, yet a strong presence of reversible lanes in the city center.