Search research articles

Search research articles

Related Experiment Video

Updated: Jun 12, 2026

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Published on: February 5, 2021

SMC, a simple method to rapidly assemble multiple fragments into one construct.

Guihong Tan¹, Change Tan

¹Division of Biological Sciences, Bond Life Sciences Center, University of Missouri, MO 65211, USA. tanc@missouri.edu

Frontiers in Bioscience (Elite Edition)

|June 3, 2010

Summary

This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Global genetic interaction network of a human cell maps conserved principles and informs functional interpretation of gene co-essentiality profiles.

Cell·2026

Same author

Domestication drives repeated evolution of sexual-asexual life cycle trade-offs in yeast.

Proceedings of the National Academy of Sciences of the United States of America·2026

Same author

A global genetic interaction map of a human cell reveals conserved principles of genetic networks.

bioRxiv : the preprint server for biology·2025

Same author

Direct structural analysis of a single acyl carrier protein domain in fatty acid synthase from the fungus Saccharomyces cerevisiae.

Communications biology·2024

Same author

The proteomic landscape of genome-wide genetic perturbations.

Cell·2023

Same author

Author Correction: Chemical genomics with pyrvinium identifies C1orf115 as a regulator of drug efflux.

Nature chemical biology·2022

Same journal

Enhancement of Cassava (<i>Manihot esculenta</i> Crantz) Retting Efficiency Using Pectinolytic and Cellulolytic <i>Streptomyces</i> sp. in the Traditional Starter Sta_96.

Frontiers in bioscience (Elite edition)·2026

Same journal

From Zinc to Manganese: Expanding the Functional Landscape of ZIP Family Metal Transporters.

Frontiers in bioscience (Elite edition)·2026

Same journal

Chimeric Antigen Receptor-Macrophages in Pancreatic Cancer: Promise, Pitfalls, and the Path Ahead.

Frontiers in bioscience (Elite edition)·2026

Same journal

<i>Lactobacillus paragasseri</i> lg94 With High L-Lactic Acid Production and Probiotic Potential.

Frontiers in bioscience (Elite edition)·2026

Same journal

Biotechnologically Derived Materials as Drug Delivery Systems for Tissue Regeneration.

Frontiers in bioscience (Elite edition)·2026

Same journal

Endolysins and Their Potential in Combating Bacterial Multidrug Resistance.

Frontiers in bioscience (Elite edition)·2026

See all related articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Split-marker-mediated multiple-piece cloning (SMC) enables rapid assembly of DNA fragments in yeast. This one-step method uses homologous recombination to fuse multiple DNA pieces into a single construct efficiently.

Area of Science:

Molecular Biology
Synthetic Biology
Yeast Genetics

Background:

Efficient assembly of multiple DNA fragments is crucial for synthetic biology and genetic engineering.
Existing methods for DNA assembly can be time-consuming and complex.

Purpose of the Study:

To develop a simple and rapid method for assembling multiple DNA fragments into a single construct in yeast.
To introduce split-marker-mediated multiple-piece cloning (SMC) as an efficient cloning strategy.

Main Methods:

Utilized a split selectable marker, with one half on a plasmid backbone.
Employed homologous recombination in yeast to reconstitute the marker gene.
Fused DNA fragments with approximately 30 base pair overlaps in a one-step process.

More Related Videos

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

Published on: February 9, 2015

Protocols for C-Brick DNA Standard Assembly Using Cpf1

Protocols for C-Brick DNA Standard Assembly Using Cpf1

Published on: June 15, 2017

Related Experiment Videos

Last Updated: Jun 12, 2026

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Published on: February 5, 2021

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

Published on: February 9, 2015

Protocols for C-Brick DNA Standard Assembly Using Cpf1

Protocols for C-Brick DNA Standard Assembly Using Cpf1

Published on: June 15, 2017

Main Results:

Successfully demonstrated the SMC method for assembling multiple DNA fragments in yeast.
The method allows for rapid, one-step fusion of DNA fragments using PCR and/or restriction enzyme-digested products.
Reconstitution of the selectable marker drives the assembly process.

Conclusions:

Split-marker-mediated multiple-piece cloning (SMC) provides a simple and rapid approach for DNA assembly in yeast.
This method facilitates the construction of complex DNA molecules efficiently.
SMC is a valuable tool for researchers in molecular biology and synthetic biology.