ArticleCell reports methods2023
An efficient cloning method to expand vector and restriction site compatibility of Golden Gate Assembly.
Article in Cell reports methods, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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6 citing papers in PubMed, 14 citations in OpenAlex.
- Advances in large DNA fragment assembly for microbial cell factory engineering.Quantitative biology (Beijing, China) · 2026Review
- One-pot cloning and protein expression platform for genetic engineering.bioRxiv : the preprint server for biology · 2025Article
- UniClo: scarless hierarchical DNA assembly without sequence constraint.Nucleic acids research · 2025Article
- Article
- Navigating the challenges of engineering composite specialized metabolite pathways in plants.The Plant journal : for cell and molecular biology · 2025Review
- Article
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2 authors at 1 institution in 1 country.
Funding
Abstract
Golden Gate Assembly is an efficient and rapid cloning method but requires dedicated vectors. Here, we modified Golden Gate to expand its compatibility to a broader range of destination vectors while maintaining its strengths. Our Expanded Golden Gate (ExGG) assembly adds to the insert(s) type IIS restriction sites that generate protruding ends compatible with traditional type IIP sites on the recipient vector. The ligated product cannot be cleaved again, owing to a single-base change near the junction. This allows the reaction to proceed in a single tube without an intermediate purification step. ExGG can be used to introduce multiple fragments into a vector simultaneously, including shorter fragments (<100 bp) and fragments with shared sequences, which can be difficult to assemble with other fast cloning strategies. Thus, ExGG extends the convenience of Golden Gate to a much larger space of pre-existing vectors designed for conventional cloning.
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