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

Figure 1.

Figure 1

Design features of SyneBrick vectors for gene expression in S. elongatus PCC 7942. (A) SyneBrick vector sections: promoters, antibiotic markers, chromosomal integrational sites and gene of interests (GOI). Standard SyneBrick vector (pSe1Bb1s-GFP) features are two homologous regions of neutral site I (NSIa and NSIb) for chromosomal integration, gene expression of a target under control of the inducible trc promoter and LacI repressor, a transcriptional terminator, spectinomycin-resistance selection, and replication of origin (pUC19) for cloning in E. coli. Sections are selected to construct a SyneBrick vector. For expansion of SyneBrick vectors, two promoters (TetA and T7 promoters), two antibiotic makers (kanamycin or chloramphenicol) and neutral sites for chromosomal integration (site II or site III) are available as interchangeable parts for the standard SyneBrick vector. (B) The nomenclature for the library of SyneBrick vector library is described. SyneBrick vectors constructed in this work are in Table 1. (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, 3, and 4 are shown in gel images. DNA sequences were verified.