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. 2017 Mar 2;8:293. doi: 10.3389/fpls.2017.00293

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Copyright © 2017 Kim, Lee, Um, Sim and Woo.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Design features of SyneBrick vectors 2.0 for gene expression using the T7 promoter in S. elongatus PCC 7942. (A) Scheme for genetic control of target gene (e.g. eYFP). With IPTG, T7 RNA polymerase (T7 TNAP) was expressed and its cognate T7 promoter was used to express target genes. Sequences of original T7 promoter and sequence variants (T7.3 and T7.4) with binding and strength regions (Temme et al., 2012). (B) Construction of cyanobacterial strains using pSe2Bb7k-eYFP, pSe2Bb7.3k-eYFP, pSe2Bb7.4k-eYFP, and pSe3Bb1c-T7RNAP vectors for expression of eYFP protein under control of T7 variant promoters. Sequencing primers are in Table S1. (C) Gel images of recombinant cyanobacterial strains. Strains were verified by PCR using oligonucleotides (arrows). Sequences are in Table S1. DNA fragments from wild-type and strains 1, 2, and 3 are shown in gel images. DNA sequences were verified.