Sequencing of the insert of the final construct used to generate the ∆<i>rv1877 </i>M.tb mutant

Here, the fragment of pGOAL17 containing the<i> sacB </i>and <i>LacZ</i> has been cloned into the p2NIL-1877US/DS construct, and this was sequenced again to check for integrity. Since the insert is large, many sequencing reactions were performed, using different primers, named primer -1 to primer 9. Therefore, alignments were performed with all data, to ensure coverage of the entire region.The last two files are the in silico designed constructs, used as template for screening by restriction enzyme digestion (RED) and for the sequencing alignments. Since cloning the fragment from pGOAL17 could face either direction, since it was a blunt end cloning, there are two possible designs of the final construct.将携带<i>sacB</i>与<i>LacZ</i>的pGOAL17片段克隆至p2NIL-1877US/DS构建体中，随后对该重组构建体进行重新测序以验证其完整性。由于插入片段体量较大，本次实验使用了命名为引物-1至引物9的多组不同引物完成了多次测序反应。因此，我们对所有测序数据进行序列比对，以确保覆盖目标区域的全长。最后两份文件为计算机模拟（in silico）设计的构建体，可用作限制性内切酶酶切（restriction enzyme digestion, RED）筛选以及测序比对的模板。由于本次克隆采用平末端克隆方式，从pGOAL17获取的插入片段可能以两种反向的方向整合入载体，因此最终构建体存在两种可行的设计方案。

搜集汇总

背景与挑战

背景概述

该数据集聚焦于结核分枝杆菌Δrv1877突变体生成过程中最终构建体的插入片段测序数据。通过克隆和多次测序反应（使用不同引物），确保了构建体的完整性验证，并涉及两种可能的构建体设计方向。数据集主要用于支持细菌学和传染病研究中的基因工程操作。