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. 2021 Sep 2;11:17626. doi: 10.1038/s41598-021-96863-x

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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In silico cloning of optimized codons encoding vaccine protein into pET28a (+) vector to ensure expression in microbial systems. The DNA sequence was inserted into the multiple cloning-site of the cloning vector. Here, the red portion denotes the gene sequence of our designed vaccine construct while the black portion denotes the backbone of the vector. All colored arrows denote the location and direction of the expression of gene. The blue portion shows vaccine codon sequence while green denotes kanamycin resistance gene, violet represents vector genes and yellow denotes origin of replication (SnapGene 5.2.5.1—https://www.snapgene.com/).