Abstract‘S16‐5503GT’ (Reg. no. CV‐555, PI 700002) is a semi‐determinate, late‐maturity group IV (relative maturity, 4.8), glyphosate‐tolerant, high‐yielding soybean [Glycine max (L.) Merr.] cultivar developed and released in 2021 by the University of Missouri‐Fisher Delta Research, Extension, and Education Center (MU‐FDREEC) soybean breeding program. Southern U.S. growers’ preference for high‐yielding, early‐maturing cultivars with glyphosate tolerance and broad disease resistance motivated the development of S16‐5503GT soybean. It was evaluated in a total of 92 environments across 12 U.S. mid‐southern states from 2017 to 2021 for yield and agronomic traits. Across all testing environments, S16‐5503GT yielded 4,258 kg ha−1, which was equivalent to 108% of the non‐Xtend check mean and 102% of the Xtend check mean, ranking in the top 19% overall. In the 2021 Southern State Variety Trials, S16‐5503GT yielded 4,455 kg ha−1 and ranked in the top 43% overall. S16‐5503GT is resistant to soybean cyst nematode races 3 (HG type 5.7) and 5 (HG type 2.5.7) and moderately resistant to race 2 (HG type 1.2.5.7). It is also resistant to southern root‐knot nematode, reniform nematode, frogeye leaf spot, charcoal rot, and brown stem rot and is tolerant to extreme salinity conditions. The broad disease resistance package combined with high‐yielding performance and wide adaptability across multiple mid‐southern states make S16‐5503GT an excellent cultivar choice for soybean growers in the southern United States, where late‐maturity group IV varieties are in demand.