ArticlePloS one2020
Enabling one-pot Golden Gate assemblies of unprecedented complexity using data-optimized assembly design.
Article in PloS one, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 61 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
61 citing papers in PubMed, 103 citations in OpenAlex.
- Megascale recombination generates millions of catalytically competent PETases.bioRxiv : the preprint server for biology · 2026Article
- Article
- The plant Golden Gate toolkit: compatibility, interoperability, and harmonized assembly in plant synthetic biology.Plant methods · 2026Review
- PrimerWeaver: an integrated web server for primer design in molecular biology workflows.Nucleic acids research · 2026Article
- A portable Cas6f-based system for multiplex translational repression in bacteria.Nature communications · 2026Article
- Scalable and cost-efficient custom gene library assembly from oligopools.Science advances · 2026Article
- Single amino acid substitution in DNA Polymerase I dramatically alters infection dynamics of bacteriophage T7.bioRxiv : the preprint server for biology · 2026Article
- Mitigating protocatechuic acid toxicity in Vibrio natriegens enables poly-3-hydroxybutyrate production from an aromatic carbon source.Applied microbiology and biotechnology · 2026Article
- Deep-mutational scanning libraries using Tiled-Region Exchange mutagenesis.G3 (Bethesda, Md.) · 2026Article
- CyanoConstruct: simple platform for cyanobacterial expression construct assembly and translational tuning.BMC biotechnology · 2026Article
- Article
- A fully synthetic Golden Gate assembly system for engineering aProceedings of the National Academy of Sciences of the United States of America · 2026Article
- Recoded Bacteriophage Genome for Bio-Orthogonal-Enabled Concentration and Detection ofACS synthetic biology · 2026Article
- High-throughput yeast engineering in biofoundries: towards autonomous and scalable synthetic biology.FEMS yeast research · 2026Review
- CloneCoordinate: Open-Source Software for Collaborative DNA Construction.ACS synthetic biology · 2025Article
- Construction and Diversification of Natural Product Biosynthetic Gene Clusters at High Efficiency and Accuracy.ACS synthetic biology · 2025Article
- Genome synthesis, assembly, and rebooting of therapeutically useful high G+C% mycobacteriophages.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- One-pot cloning and protein expression platform for genetic engineering.bioRxiv : the preprint server for biology · 2025Article
- Expanding genetic engineering capabilities in Vibrio natriegens with the Vnat Collection.Nucleic acids research · 2025Article
- Combining Machine Learning and Multiplexed,bioRxiv : the preprint server for biology · 2025Article
1 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
DNA assembly is an integral part of modern synthetic biology, as intricate genetic engineering projects require robust molecular cloning workflows. Golden Gate assembly is a frequently employed DNA assembly methodology that utilizes a Type IIS restriction enzyme and a DNA ligase to generate recombinant DNA constructs from smaller DNA fragments. However, the utility of this methodology has been limited by a lack of resources to guide experimental design. For example, selection of the DNA sequences at fusion sites between fragments is based on broad assembly guidelines or pre-vetted sets of junctions, rather than being customized for a particular application or cloning project. To facilitate the design of robust assembly reactions, we developed a high-throughput DNA sequencing assay to examine reaction outcomes of Golden Gate assembly with T4 DNA ligase and the most commonly used Type IIS restriction enzymes that generate three-base and four-base overhangs. Next, we incorporated these findings into a suite of webtools that design assembly reactions using the experimental data. These webtools can be used to create customized assemblies from a target DNA sequence or a desired number of fragments. Lastly, we demonstrate how using these tools expands the limits of current assembly systems by carrying out one-pot assemblies of up to 35 DNA fragments. Full implementation of the tools developed here enables direct expansion of existing assembly standards for modular cloning systems (e.g. MoClo) as well as the formation of robust new high-fidelity standards.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.