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Updated: Jul 4, 2026

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
Published on: February 5, 2021
Modular construction of plasmids through ligation-free assembly of vector components with oligonucleotide linkers
Jonathan A Vroom1, Clifford L Wang
1Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA.
Abstract:
We have developed a modular method of plasmid construction that can join multiple DNA components in a single reaction. A nicking enzyme is used to create 5' and 3' overhangs on PCR-generated DNA components. Without the use of ligase or restriction enzymes, components are joined using oligonucleotide linkers that recognize the overhangs. By specifying the sequences of the linkers, desired components can be assembled in any combination and order to generate different plasmid vectors.
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