The 2A sequence can be used to successfully and predictably target proteins to distinct subcellular compartments. Various targeting sequences were N-terminally fused to an upstream vYFP and a downstream tdTom reporter separated by T2A. The vYFP and tdTom proteins were localized using direct fluorescence microscopy imaging. Mitochondria were stained with MitoTracker (MT), and nuclei with Hoechst 33342. Legend: ATS = apicoplast targeting sequence; MTS = mitochondrial targeting sequence and PEX = protein export element.