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Updated: Aug 13, 2025

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
Published on: February 5, 2021
Easy method for six-fragment Golden Gate Assembly of modular vectors
Ionut Adrian Cepleanu-Pascu1, Miruna Stan2, Sebastian Cocioba3
1Department of Genetics, Faculty of Biology, University of Bucharest, Bucharest, Romania.
Abstract:
Efficient cloning techniques are a requirement for synthetic biology. This study provides a simplified cloning method based on Golden Gate Assembly that can be used for rapid vector construction. The building of multiple expression vectors with customizable modules is achieved in a timely manner with minimal hands-on time by removing unnecessary steps in the workflow. The authors constructed a total of 21 mammalian episomal expression vectors and conducted a fluorescence expression comparison for different regulatory region combinations post-transfection in HEK293T and HEPG2 cells. Screening revealed that using the EF-1α promoter in combination with the bovine growth hormone polyadenylation sequence seemed to perform best in the types of cells tested compared with other variants.
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