| Summary: | <i>Lactococcus lactis</i> is a commonly used fermenting bacteria in cheese, beverages and meat products. Due to the lack of simplified chassis strains, it has not been widely used in the fields of synthetic biology. Thus, the construction of lactic acid bacteria chassis strains becomes more and more important. In this study, we performed whole genome sequencing, annotation and analysis of <i>L. lactis</i> N8. Based on the genome analysis, we found that <i>L. lactis</i> N8 contains two large plasmids, and the function prediction of the plasmids shows that some regions are related to carbohydrate transport/metabolism, multi-stress resistance and amino acid uptake. <i>L. lactis</i> N8 contains a total of seven prophage-related fragments and twelve genomic islands. A gene cluster encoding a hybrid NRPS–PKS system that was found in <i>L. lactis</i> N8 reveals that the strain has the potential to synthesize novel secondary metabolites. Furthermore, we have constructed a simplified genome chassis of <i>L. lactis</i> N8 and achieved the largest amount of deletion of <i>L. lactis</i> so far. Taken together, the present study offers further insights into the function and potential role of <i>L. lactis</i> N8 as a model strain of lactic acid bacteria and lays the foundation for its application in the field of synthetic biology.
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