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. 2016 Jul 2;13:119. doi: 10.1186/s12985-016-0574-7

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© The Author(s). 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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.

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Fig. 1 — Schematic presentation of plasmids for run-off RNA transcription and protein expression. a cDNA of full genome segments were cloned between the promoter for DNA dependent RNA polymerase of bacteriophage T7 (P_T7) and a recognition site of restriction enzyme (RE) SapI, BsmBI, or BsaI to allow run-off RNA transcription. The appropriate RE was selected based on the absence of the recognition sites in the respective genome segment. Alternatively, internal sites were eliminated by silent mutations with respect to the translated amino acid sequence. b Authentic or open reading frames (ORFs) optimized for plasmid stability and eukaryotic expression (see materials and methods) were cloned between the immediate early promoter of human cytomegalovirus (P_HCMV) and the polyA signal (polyA) and transcription terminator (TER)