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. 2022 Dec 22;51(1):271–289. doi: 10.1093/nar/gkac1175

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© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Luciferase reporter assays of native initiation sequences harboring potentially ambiguous start sites. To create an authentic initiation context, 20 nts of the native RBS and 15 nts of the corresponding CDS were fused N-terminally to the luciferase reporter CDS. Relative light units (RLU) measured from constructs harboring the fLUC CDS in frame of either potential start codon are displayed for the E. coli genes hdhA (A), secF (B), pqiA (C), tamA (D), asnB (E) and nagZ (F). The start codon in frame of the luciferase gene is indicated in upper case and bold. RLU measured from constructs harboring the wild type (wt) sequences are indicated in white, from the alternative ORF in light grey and from controls with distinct start codons in dark grey. The wt RBS sequences containing the AUGUG or GUGUG start sites are displayed below the respective graphs. The predicted SD sequences are depicted in bold and the determined start codons are underlined. The mean and the standard deviation of the independent measurements are provided.