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Updated: Jun 11, 2025

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
Published on: February 5, 2021
Construction of Small Genes with Repetitive Elements Using Oligo Extension and Golden Gate Assembly
Michael T A Nguyen1,2, Ebbe Sloth Andersen3,4, Georgios Pothoulakis4
1Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT, USA.
Abstract:
Gene synthesis efficiency has greatly improved in recent years but is limited when it comes to repetitive sequences and results in synthesis failure or delays by DNA synthesis vendors. Here, we describe a method for the assembly of small synthetic genes with repetitive elements: First, a gene of interest is split in silico into small synthons of up to 80 base pairs flanked by Golden Gate-compatible overhangs. Then synthons are made by oligo extension and finally assembled into a synthetic gene by Golden Gate assembly.
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