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The unicellular Synechocystis sp. PCC 6803, already considered the \u2018green\u2019 Escherichia coli, is the best studied cyanobacterium but to be used as an efficient and robust photoautotrophic chassis it requires a customized and well-characterized toolbox. In this context, we evaluated the possibility of using three self-replicative vectors from the Standard European Vector Architecture (SEVA) repository to transform Synechocystis. Our results demonstrated that the presence of the plasmid does not lead to an evident phenotype or hindered Synechocystis growth, being the vast majority of the cells able to retain the replicative plasmid even in the absence of selective pressure. In addition, a set of heterologous and redesigned promoters were characterized exhibiting a wide range of activities compared to the reference PrnpB, three of which could be efficiently repressed. As a proof-of-concept, from the expanded toolbox, one promoter was selected and assembled with the ggpS gene [encoding one of the proteins involved in the synthesis of the native compatible solute glucosylglycerol (GG)] and the synthetic device was introduced into Synechocystis using one of the SEVA plasmids. The presence of this device restored the production of the GG in a ggpS deficient mutant validating the functionality of the tools\/device developed in this study.<\/jats:p>","DOI":"10.1093\/synbio\/ysy014","type":"journal-article","created":{"date-parts":[[2018,8,8]],"date-time":"2018-08-08T12:33:19Z","timestamp":1533731599000},"source":"Crossref","is-referenced-by-count":49,"title":["Expanding the toolbox for Synechocystis sp. PCC 6803: validation of replicative vectors and characterization of a novel set of promoters"],"prefix":"10.1093","volume":"3","author":[{"given":"Eunice A","family":"Ferreira","sequence":"first","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto, Porto, Portugal"},{"name":"IBMC \u2013 Instituto de Biologia Molecular e Celular Universidade do Porto, Porto, Portugal"},{"name":"ICBAS \u2013 Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar Universidade do Porto, Porto, Portugal"}]},{"given":"Catarina C","family":"Pacheco","sequence":"additional","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto, Porto, Portugal"},{"name":"IBMC \u2013 Instituto de Biologia Molecular e Celular Universidade do Porto, Porto, Portugal"}]},{"given":"Filipe","family":"Pinto","sequence":"additional","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto, Porto, Portugal"},{"name":"IBMC \u2013 Instituto de Biologia Molecular e Celular Universidade do Porto, Porto, Portugal"},{"name":"School of Biological Sciences, University of Edinburgh, Edinburgh, UK"},{"name":"Centre for Synthetic and Systems Biology, University of Edinburgh, Edinburgh, UK"}]},{"given":"Jos\u00e9","family":"Pereira","sequence":"additional","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto, Porto, Portugal"},{"name":"IBMC \u2013 Instituto de Biologia Molecular e Celular Universidade do Porto, Porto, Portugal"}]},{"given":"Pedro","family":"Lamosa","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, ITQB NOVA, Oeiras, Portugal"}]},{"given":"Paulo","family":"Oliveira","sequence":"additional","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto, Porto, Portugal"},{"name":"IBMC \u2013 Instituto de Biologia Molecular e Celular Universidade do Porto, Porto, Portugal"}]},{"given":"Boris","family":"Kirov","sequence":"additional","affiliation":[{"name":"CNRS-UMR8030 Laboratoire iSSB and Universit\u00e9 Paris-Saclay and Universit\u00e9 d\u2019\u00c9vry and CEA DRF, IG, Genoscope, \u00c9vry, France"},{"name":"ANP - Faculty of Automatics, TU \u2013 Sofia, Sofia, Bulgaria"},{"name":"BioInfoTech Lab - RDIC, Sofia Tech Park, Sofia, Bulgaria"}]},{"given":"Alfonso","family":"Jaramillo","sequence":"additional","affiliation":[{"name":"CNRS-UMR8030 Laboratoire iSSB and Universit\u00e9 Paris-Saclay and Universit\u00e9 d\u2019\u00c9vry and CEA DRF, IG, Genoscope, \u00c9vry, France"},{"name":"Warwick Integrative Synthetic Biology Centre and School of Life Sciences, University of Warwick, Coventry, UK"},{"name":"Institute for Integrative Systems Biology (I2SysBio) University of Valencia-CSIC, Paterna, Spain"}]},{"given":"Paula","family":"Tamagnini","sequence":"additional","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto, Porto, Portugal"},{"name":"IBMC \u2013 Instituto de Biologia Molecular e Celular Universidade do Porto, Porto, Portugal"},{"name":"Faculdade de Ci\u00eancias, Departamento de Biologia, Universidade do Porto, Porto, Portugal"}]}],"member":"286","published-online":{"date-parts":[[2018,8,8]]},"reference":[{"key":"2020081109471553100_ysy014-B1","first-page":"1","volume-title":"The Cyanobacteria: Molecular Biology, Genomics, and Evolution","author":"Knoll","year":"2008"},{"key":"2020081109471553100_ysy014-B2","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1093\/dnares\/3.3.109","article-title":"Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC 6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions","volume":"3","author":"Kaneko","year":"1996","journal-title":"DNA Res"},{"key":"2020081109471553100_ysy014-B3","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1002\/biot.201000109","article-title":"Flux coupling and transcriptional regulation within the metabolic network of the photosynthetic bacterium Synechocystis sp. PCC 6803","volume":"6","author":"Montagud","year":"2011","journal-title":"Biotechnol. J"},{"key":"2020081109471553100_ysy014-B4","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1002\/jctb.2065","article-title":"Genome-scale modeling of Synechocystis sp. PCC 6803 and prediction of pathway insertion","volume":"84","author":"Fu","year":"2009","journal-title":"J. Chem. Technol. 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