MOTIVATION: The evolution and role of gene clusters in eukaryotes is poorly understood. Currently, most studies and computational prediction programs limit their focus to specific types of clusters, such as those involved in secondary metabolism. RESULTS: We present EvolClust, a python-based tool for the inference of evolutionary conserved gene clusters from genome comparisons, independently of the function or gene composition of the cluster. EvolClust predicts conserved gene clusters from pairwise genome comparisons and infers families of related clusters from multiple (all versus all) genome comparisons. AVAILABILITY AND IMPLEMENTATION: https://github.com/Gabaldonlab/EvolClust/. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.