Optimized vectors for genetic engineering of <i>Aureobasidium pullulans</i>

<i>Aureobasidium pullulans</i> is a polyextremotolerant black yeast that exhibits impressive morphological plasticity. Consequently, it shows promise as a model system for investigating mechanisms of cell adaptation to different environments and the regulation of cell shape. Here, we build upon the current toolkit for working with <i>A. pullulans</i> and design and test 25 vectors with seven different codon-optimized fluorophores and three selection cassettes. This includes vectors that allow for dual expression of green fluorescent protein and mCherry-tagged proteins at the <i>URA3</i> locus and vectors that enable homology-based deletion or C-terminal tagging of endogenous genes without the need for cloning. This versatile vector series for working with <i>A. pullulans</i> will enable a broad range of experiments in this emerging model system.