The files in this collection consist of the improved genomic sequences, gene annotations, and evidence tracks that were used in the analyses described in the manuscript “Drosophilia Muller F Elements Maintain a Distinct Set of Genomic Properties Over 40 Million Years of Evolution” (doi: 10.1534/g3.114.015966). The improved genomic sequences and gene annotations were produced by students participating in the Genomics Education Partnership (GEP; http://gep.wustl.edu) during 2007–2012. This study seeks to understand the evolution of the Muller F element through the comparative analysis of Drosophila melanogaster, Drosophila erecta, Drosophila mojavensis, and Drosophila grimshawi. As part of this analysis, we have improved the genomic scaffolds from the D. mojavensis and D. grimshawi assemblies, and produced manual gene annotations for D. erecta, D. mojavensis, and D. grimshawi. The digital assets include the improved sequences and annotations in GenBank ASN.1 format, and the files required to create the UCSC Genome Browser instances for D. melanogaster, D. erecta, D. mojavensis, and D. grimshawi. The Genome Browser instances include the improved genomic sequences, gene annotations, and the results of the bioinformatics analyses described in the manuscript.