Abstract: The US Army Engineer Research and Development Center, Coastal and Hydraulics Laboratory (CHL), used the Ship/Tow Simulator to evaluate navigational conditions for the US Army Corps of Engineers, St. Louis District (MVS), proposed 1,200 feet (ft) lock chamber at Lock and Dam 25 in a tow simulation study. The study considers the impacts to navigation throughout construction sequences of the proposed 1,200 ft lock chamber and the final completed project. Testing occurred at CHL in October–November 2022 with five industry tow pilots. A total of 47 unique test conditions for a total of 187 ship-simulation exercises were evaluated. All final project simulations indicated that the design is feasible. When testing the construction scenarios of the design, it was evident that a tug assist boat would be necessary for entering the 600 ft lock for both approaches. Results found that the intermediate wall construction should begin at the existing structure and progress downstream. Entering the 600 ft lock from the pool side was additionally completed successfully; however, modifications are needed for entering from the tailwater side. Ultimately, the results of this study will aid MVS in the design plan and decision-making regarding the proposed lock.