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. 2017 Sep 5;8(5):e01291-17. doi: 10.1128/mBio.01291-17

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Copyright © 2017 Zhang et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 1 — Direct cloning and proteobacterial broad-host-range heterologous expression of microbial BGCs. (A) Organization of TAR cloning and broad-host-range expression vector pCAP05 composed of elements for maintenance (CEN6/ARS4), selection (HIS3), and counterselection (pAHD1, URA3) in yeast (blue) and broad-host-range maintenance (oriV, trfA) and selection (tetA, tetR) in Gram-negative bacteria (tan). Additionally, incC and korA1 regulate plasmid maintenance and oriT and traJ enable conjugal transfer of pCAP05. The vector can be linearized for cloning by digestion with ClaI and XhoI. (B) Schematic illustration of the broad-host-range heterologous expression concept. A generic microbial BGC cloned into pCAP05 is depicted above three proteobacterial expression hosts that were selected and validated in this study.