GENOMICON-Seq is a comprehensive genomic sequencing simulation tool that enables the assessment of laboratory and bioinformatics parameters influencing the detection of mutations. The tool generates genomes with mutations, mimicking processes such as low-frequency mutations, APOBEC3 activity in viruses, somatic mutations and single base substitution (SBS) mutational signals. GENOMICON-Seq adds amplicon and whole exome sequencing biases and errors. It outputs sequencing reads compatible with mutation detection tools and a report on mutation origin (generated mutations and PCR errors), nucleotide context, and position. GENOMICON-Seq aids in the evaluation of bioinformatics tools and experimental designs, reducing the need for costly real-world sequencing experiments.