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The Golden Gate cloning technique is used to assemble DNA parts into higher-order assemblies. Individual parts containing compatible overhangs generated by type IIS restriction enzymes are joined together using DNA ligase. The technique enables users to assemble custom transcription units (TUs) for a wide array of experimental assays. Several Golden Gate cloning systems have been developed; however, they are typically used with a narrow range of organisms. Here we describe the Multi-Kingdom (MK) cloning system that allows users to generate DNA plasmids for use in a broad range of organisms.

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Assembling DNA Plasmids with the Multi-Kingdom (MK) Cloning System

  • 实验方案

  • 发表日期：

    2025年

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  • Sarina Scoville,
  • David M. Chiasson

摘要

The Golden Gate cloning technique is used to assemble DNA parts into higher-order assemblies. Individual parts containing compatible overhangs generated by type IIS restriction enzymes are joined together using DNA ligase. The technique enables users to assemble custom transcription units (TUs) for a wide array of experimental assays. Several Golden Gate cloning systems have been developed; however, they are typically used with a narrow range of organisms. Here we describe the Multi-Kingdom (MK) cloning system that allows users to generate DNA plasmids for use in a broad range of organisms.