The replication of DNA is a complex biological process that is essential for life. BacterialDNA replicationis initiated at genomic loci referred to asreplication origins(oriCs). Integrating theZ-curvemethod, DnaA box distribution, and comparative genomic analysis, we developed a web server to predict bacterialoriCs in 2008 called Ori-Finder, which is helpful to clarify the characteristics of bacterialoriCs. TheoriCs of hundreds of sequenced bacterial genomes have been annotated in the genome reports using Ori-Finder and the predicted results have been deposited in DoriC, a manually curated database oforiCs. This has facilitated large-scale data mining of functional elements inoriCs and strand-biased analysis. Here, we describe Ori-Finder 2022 with updated prediction framework, interactive visualization module, new analysis module, and user-friendly interface. More species-specific indicator genes and functional elements oforiCs are integrated into the updated framework, which has also been redesigned to predictoriCs in draft genomes. The interactive visualization module displays more genomic information related tooriCs and their functional elements. The analysis module includes regulatory protein annotation, repeat sequence discovery, homologousoriCsearch, and strand-biased analyses. The redesigned interface provides additional customization options fororiCprediction. Ori-Finder 2022 is freely available athttp://tubic.tju.edu.cn/Ori-Finder/andhttps://tubic.org/Ori-Finder/.