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. 2021 Jun 27;14:146. doi: 10.1186/s13068-021-02000-1

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© The Author(s) 2021

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 7 — Schematic diagram of plasmids and promoter replacement method. The constructed pEZ_EGFP (a), long fragment for replacing the native promoter of target gene on the genome (b), and pEZ_Target (Target means one of these three constructs: clpB, an operon containing clpB_groESL with groESL driven by their native promoter, and ZMO0994) (c). P_lacUV5 is a constitutive promoter. rrnB T1 and T7Te are terminators. Zymo Ori means an origin of replication with promoters for Z. mobilis. p15A Ori means an origin of replication with promoters for E. coli. P refers to the original promoter of the gene of interest. US means the upstream of the original promoter. aadA refers to spectinomycin resistance gene. P_tet is a bi-directional promoter, which simultaneously initiates the expression of tetR gene and the candidate gene. P_tet fragment included aadA, tetR, and P_tet