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Allelic exchange is a technique that relies on homologous recombination to replace native loci with engineered sequences. However, current allelic-exchange vectors often enable only weak selection for successful homologous recombination. We developed a suite of new allelic-exchange vectors, pTOX, which were validated in several medically important proteobacteria. They encode visible nonfluorescent chromoproteins that enable easy identification of colonies bearing integrated vectors and permit stringent selection for the second step of homologous recombination. We demonstrate the utility of these vectors by using them to investigate the effect of inactivation of\n                    <jats:named-content content-type=\"genus-species\">Serratia marcescens<\/jats:named-content>\n                    peptidoglycan amidohydrolases on \u03b2-lactam antibiotic resistance.\n                  <\/jats:p>","DOI":"10.1128\/aem.00990-19","type":"journal-article","created":{"date-parts":[[2019,6,12]],"date-time":"2019-06-12T08:50:38Z","timestamp":1560329438000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":63,"title":["A New Suite of Allelic-Exchange Vectors for the Scarless Modification of Proteobacterial Genomes"],"prefix":"10.1128","volume":"85","author":[{"given":"Jacob E.","family":"Lazarus","sequence":"first","affiliation":[{"name":"Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA"},{"name":"Department of Microbiology, Harvard Medical School, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alyson R.","family":"Warr","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Harvard Medical School, Boston, Massachusetts, USA"},{"name":"Division of Infectious Diseases, Brigham and Women's Hospital, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carole J.","family":"Kuehl","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Harvard Medical School, Boston, Massachusetts, USA"},{"name":"Division of Infectious Diseases, Brigham and Women's Hospital, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel T.","family":"Giorgio","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Harvard Medical School, Boston, Massachusetts, USA"},{"name":"Division of Infectious Diseases, Brigham and Women's Hospital, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brigid M.","family":"Davis","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Harvard Medical School, Boston, Massachusetts, USA"},{"name":"Division of Infectious Diseases, Brigham and Women's Hospital, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew K.","family":"Waldor","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Harvard Medical School, Boston, Massachusetts, USA"},{"name":"Division of Infectious Diseases, Brigham and Women's Hospital, Boston, Massachusetts, USA"},{"name":"Howard Hughes Medical Institute, Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1002\/0471142727.mb0116s78"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2508"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01248-15"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep15096"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.179.20.6228-6237.1997"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.170.6.2575-2583.1988"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.9.4011-4017.1998"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.145.2.1110-1111.1981"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2015.115"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt1075"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv248"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00017-11"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.174.19.6125-6137.1992"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(82)90106-8"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.plasmid.2004.02.003"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1021\/acssynbio.6b00030"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt1139"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.2144\/000112112"},{"key":"e_1_3_3_20_2","first-page":"148","article-title":"DNA tuner: a computer program for the construction of polylinker sequences of molecular vectors","volume":"9","author":"Latynski US","year":"2014","unstructured":"Latynski US, Valentovich LN. 2014. 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