Mucormycosis is a life-threatening disease especially to individuals who are immunosuppressed or have chronic health conditions. Several genera within the fungal order Mucorales have been identified as causative agents. This dataset is the result of research elucidating the population genetics, phylogeny, and pathogenicity of the infectious fungi. Included are de novo genome sequences and annotations for 27 isolates of 15 species of the following genera: Apophysomyces, Cokeromyces, Cunninghamella, Mucor, Rhizomucor, Rhizopus, Saksenaea, Syncephalastrum, and Umbelopsis. With the inclusion of previously published genomes of additional strains, comparative analysis was conducted on a total of 41. To investigate Mucorales fungi-host cell responses, RNA sequencing was performed using human airway epithelial cells infected with Rhizopus delemar or Rhizopus oryzae. All sequences are available in the NCBI SRA database with additional data provided via supplementary files.