The eastern Rhine Graben Boundary Fault (eastern RGBF) forms the eastern margin of the Upper Rhine Graben (URG), the most seismically active area in the plate interiors of Europe. Despite seismic activity posing a significant threat to the densely populated URG and critical facilities therein, only a few studies have documented the paleoearthquake history and associated seismic hazard, focusing mainly on the western margin. We present the results of the first paleoseismological trenching ever conducted on the central section of the eastern RGBF. Following high-resolution near-surface geophysical studies, we excavated six trenches near Ettlingen-Oberweier (south of Karlsruhe, Germany) on one of its secondary fault strands. The nearly-vertical fault is transtensional left-lateral and splits into several NNW-SSE en échelon branches, forming a negative flower structure. Stratigraphic and structural relationships along with radiocarbon and Optically Stimulated Luminescence dating reveal a minimum of three surface-rupturing paleoearthquakes with a Mw of potentially 6.5 ± 0.5, occurring from old to young, >56 ka (EX), between 54 and 19 ka (EY), and between 15 and 1 ka (EZ). The events are poorly constrained in age due to erosional unconformities. Based on the cumulative vertical separation of 1.2 ± 0.3 m, we calculate an average vertical slip rate of 0.02 ± 0.005 mm/yr. From a horizontally offset alluvial channel, we infer a cumulative left-lateral slip of 5.9 ± 0.7 m and derive an average horizontal slip rate of 0.1 ± 0.01 mm/yr. The average net slip rate is 0.1 ± 0.02 mm/yr for the past 59.5 ± 3.8 ka. This value represents a minimum slip rate for the eastern RGBF, considering slip distribution within the different fault strands of the fault system. Our findings highlight the seismic potential of the eastern RGBF, providing new evidence of the Late Pleistocene and Holocene tectonic activity of its central section.