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Among them, we take the fast\u2010growing cyanobacterium\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Synechococcus elongatus<\/jats:italic>\n                    <\/jats:styled-content>\n                    UTEX 2973 (Syn2973) as a representative study due to its promising roles for CO\n                    <jats:sub>2<\/jats:sub>\n                    fixation and bioconversion. We established three marker\u2010free platforms to achieve stable genome recombination: (i) T4CROSS, which employs two plasmids and four rounds of single crossover; (ii) TRIPLEARM, which uses a single plasmid containing three homologous arms for three rounds of single crossover; and (iii) CRISPRARM, which integrates CRISPR\/Cpf1\u2010mediated genome editing with homologous recombination. As proof of concept, we employed the CRISPRARM platform for a three\u2010step sequential engineering of the sucrose biosynthetic pathway. The final engineered strain produced 7.12\u2009g\u2009L\n                    <jats:sup>\u22121<\/jats:sup>\n                    of sucrose within 4\u2009days.\n                  <\/jats:p>","DOI":"10.1111\/pbi.70702","type":"journal-article","created":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T04:55:15Z","timestamp":1781585715000},"page":"5284-5299","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Iterative Genome Engineering Platform Enables Efficient Sucrose Biosynthesis From\n                    <scp>\n                      CO\n                      <sub>2<\/sub>\n                    <\/scp>\n                    in Photosynthetic\n                    <i>Synechococcus elongatus<\/i>\n                    <scp>UTEX<\/scp>\n                    2973"],"prefix":"10.1111","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1443-6342","authenticated-orcid":false,"given":"Shubin","family":"Li","sequence":"first","affiliation":[{"name":"School of Synthetic Biology and Biomanufacturing Tianjin University  Tianjin People's Republic of China"},{"name":"Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering Ministry of Education of China  Tianjin People's Republic of China"},{"name":"State Key Laboratory of Synthetic Biology Tianjin University  Tianjin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7575-3483","authenticated-orcid":false,"given":"Tao","family":"Sun","sequence":"additional","affiliation":[{"name":"School of Synthetic Biology and Biomanufacturing Tianjin University  Tianjin People's Republic of China"},{"name":"Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering Ministry of Education of China  Tianjin People's Republic of China"},{"name":"State Key Laboratory of Synthetic Biology Tianjin University  Tianjin People's Republic of China"},{"name":"Center for Biosafety Research and Strategy Tianjin University  Tianjin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dailin","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Synthetic Biology and Biomanufacturing Tianjin University  Tianjin People's Republic of China"},{"name":"Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering Ministry of Education of China  Tianjin People's Republic of China"},{"name":"State Key Laboratory of Synthetic Biology Tianjin University  Tianjin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tong","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Synthetic Biology and Biomanufacturing Tianjin University  Tianjin People's Republic of China"},{"name":"Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering Ministry of Education of China  Tianjin People's Republic of China"},{"name":"State Key Laboratory of Synthetic Biology Tianjin University  Tianjin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Chen","sequence":"additional","affiliation":[{"name":"School of Synthetic Biology and Biomanufacturing Tianjin University  Tianjin People's Republic of China"},{"name":"Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering Ministry of Education of China  Tianjin People's Republic of China"},{"name":"State Key Laboratory of Synthetic Biology Tianjin University  Tianjin People's Republic of China"},{"name":"Haihe Laboratory of Sustainable Chemical Transformations  Tianjin 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