Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT, USA.
Systems Biology Institute, Yale University, West Haven, CT, USA.
Methods Mol Biol. 2025;2850:219-227. doi: 10.1007/978-1-0716-4220-7_12.
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.
近年来，基因合成效率有了很大的提高，但在重复序列方面仍受到限制，这会导致 DNA 合成供应商合成失败或延迟。在这里，我们描述了一种用于组装具有重复元件的小合成基因的方法：首先，通过计算机将目标基因分割成小的合成子，每个合成子长 80 个碱基对，两侧带有 Golden Gate 兼容的突出端。然后通过寡核苷酸延伸合成合成子，最后通过 Golden Gate 组装将合成子组装成合成基因。
