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. 2022 Dec 9;13:7624. doi: 10.1038/s41467-022-33576-3

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a An E. lenta endogenous plasmid pEL11863 was chosen for further engineering. b Construction of shuttle plasmids pXD69m1 and pXD69m2, and the broad-host-range plasmid pXD68Kan2. Shuttle plasmids pXD69m1 and pXD69m2 were constructed by assembly of E. lenta pEL11863 replicon (dark green), E. coli ColE1 origin and bla ampicillin-resistance gene (blue), and E. lenta antibiotic resistance gene tetW (orange) or aphA (light green). pXD68Kan2 was constructed from RSF1010-based broad host-range plasmid pAM5409, with antibiotic resistance gene replaced by aphA, and the RSF1010 replicon (dark green) and yfp (grey) originally on pAM5409 were retained. c Plasmids pXD69m1, pXD69m2 and pXD68Kan2 can be transformed into E. lenta DSM 2243 using electroporation. d Plasmid-specific PCR confirmed plasmid presence within individual colonies. Plasmids (P) and WT gDNA were used as control templates. Amplified regions are indicated in Supplementary Fig. 1a. M: DNA ladder. e Plasmid pXD69m2 can be transformed into Gordonibacter sp. 28C using electroporation. f The transformation efficiency of plasmids pXD69m1, pXD69m2 and pXD68Kan2 into different E. lenta and Gordonibacter strains. ND: colonies not detected. Data in panel f are represented as mean ± SD with n = 3 biological replicates. Source data are provided as a Source Data file.