Actinomycetales, such as the genus Streptomyces, are well‐known cell factories employed to produce a wide variety of secondary metabolites for industrial use. However, not only is the genetic engineering of Streptomyces more complicated and tedious than other model laboratory species, such as Escherichia coli, there is also a considerable lack of genetic tools, hindering its adoption as a common chassis for synthetic biology. In this work, 23 novel shuttle vectors are presented that follow the canonical SEVA (Standard European Vector Architecture) common architecture with the goal of increasing the genetic toolbox repertoire for Streptomyces and other actinomycetes. The ORI module of these plasmids is composed of the combination of two origins of replication, one for Gram‐negative bacteria and the other for Streptomyces, a Gram‐positive bacteria. Origins of replication have been included in the collection for integrative, low‐copy number, and medium‐to‐high‐copy number vectors for Streptomyces. Also, a new selection marker has been developed that confers resistance to apramycin. The functionality of these plasmids was tested via the heterologous expression of GFP and the heterologous production of the plant flavonoid apigenin in Streptomyces albus J1074, with successful results in both cases, therefore expanding the current repertoire of genetic manipulation tools in Streptomyces species.

中文翻译：

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用于放线菌和革兰氏阴性细菌的多功能SEVA穿梭载体。

放线菌属，例如链霉菌属，是众所周知的细胞工厂，用于生产各种用于工业用途的次级代谢产物。然而，链霉菌的基因工程不仅比大肠杆菌等其他实验室模型物种更为复杂和繁琐，而且还缺乏遗传工具，这阻碍了其被用作合成生物学的通用框架。在这项工作中，提出了23种新颖的穿梭载体，它们遵循规范的SEVA（标准欧洲矢量体系结构）通用体系结构，目的是增加链霉菌的遗传工具箱库和其他放线菌。这些质粒的ORI模块由两个复制起点组合而成，一个复制起点用于革兰氏阴性细菌，另一个复制于链霉菌（革兰氏阳性细菌）。复制的起点已包含在链霉菌的整合，低拷贝数和中高拷贝数载体中。而且，已经开发了赋予对阿霉素抗性的新的选择标记。这些质粒的功能通过链霉菌J1074中GFP的异源表达和植物类黄酮芹菜素的异源生产进行了测试，在两种情况下均获得成功的结果，因此扩展了链霉菌中目前的遗传操作工具范围 种类。