In conclusion, we successfully synthesize a novel ZLS, DNL-16, using 1,4-MPBOH as the OSDA. The unique layered structure of DNL-16, characterized by bre CBUs, is intricately unraveled using 3D ED. Further, the precise location of 1,4-MPBOH, situated between adjacent L0 layers of DNL-16, is determined through high-quality PXRD complemented by Rietveld refinement. These findings not only reveal the complex structure of DNL-16 but also lay a foundational groundwork for future investigations into its potential applications.