title: Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units.
abstract: We have adapted the Golden Gate DNA assembly method to the assembly of transcription units (TUs) for the yeast Saccharomyces cerevisiae, in a method we call yeast Golden Gate (yGG). yGG allows for the easy assembly of TUs consisting of promoters (PRO), coding sequences (CDS), and terminators (TER). Carefully designed overhangs exposed by digestion with a type IIS restriction enzyme enable virtually seamless assembly of TUs that, in principle, contain all of the information necessary to express a gene of interest in yeast. We also describe a versatile set of yGG acceptor vectors to be used for TU assembly. These vectors can be used for low or high copy expression of assembled TUs or integration into carefully selected innocuous genomic loci. yGG provides synthetic biologists and yeast geneticists with an efficient new means by which to engineer S. cerevisiae.
authors.affiliation: †Institute for Systems Genetics, Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, 550 First Avenue, New York, New York 10016, United States.
authors.forename: Neta
authors.initials: N
authors.lastname: Agmon
authors.affiliation: †Institute for Systems Genetics, Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, 550 First Avenue, New York, New York 10016, United States.
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Mitchell
authors.affiliation: ‡High Throughput Biology Center, School of Medicine, Johns Hopkins University, Edward D. Miller Research Building, 733 North Broadway, Baltimore, Maryland 21205, United States.|§School of Biological Sciences, University of Edinburgh, The King's Buildings, Edinburgh, EH9 3JR, United Kingdom.
authors.forename: Yizhi
authors.initials: Y
authors.lastname: Cai
authors.affiliation: ‡High Throughput Biology Center, School of Medicine, Johns Hopkins University, Edward D. Miller Research Building, 733 North Broadway, Baltimore, Maryland 21205, United States.
authors.forename: Shigehito
authors.initials: S
authors.lastname: Ikushima
authors.affiliation: ‡High Throughput Biology Center, School of Medicine, Johns Hopkins University, Edward D. Miller Research Building, 733 North Broadway, Baltimore, Maryland 21205, United States.
authors.forename: James
authors.initials: J
authors.lastname: Chuang
authors.affiliation: ‡High Throughput Biology Center, School of Medicine, Johns Hopkins University, Edward D. Miller Research Building, 733 North Broadway, Baltimore, Maryland 21205, United States.
authors.forename: Allen
authors.initials: A
authors.lastname: Zheng
authors.affiliation: ‡High Throughput Biology Center, School of Medicine, Johns Hopkins University, Edward D. Miller Research Building, 733 North Broadway, Baltimore, Maryland 21205, United States.
authors.forename: Woo-Jin
authors.initials: WJ
authors.lastname: Choi
authors.affiliation: †Institute for Systems Genetics, Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, 550 First Avenue, New York, New York 10016, United States.
authors.forename: J Andrew
authors.initials: JA
authors.lastname: Martin
authors.affiliation: ‡High Throughput Biology Center, School of Medicine, Johns Hopkins University, Edward D. Miller Research Building, 733 North Broadway, Baltimore, Maryland 21205, United States.
authors.forename: Katrina
authors.initials: K
authors.lastname: Caravelli
authors.affiliation: ‡High Throughput Biology Center, School of Medicine, Johns Hopkins University, Edward D. Miller Research Building, 733 North Broadway, Baltimore, Maryland 21205, United States.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Stracquadanio
authors.affiliation: †Institute for Systems Genetics, Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, 550 First Avenue, New York, New York 10016, United States.
authors.forename: Jef D
authors.initials: JD
authors.lastname: Boeke
mesh_terms: D020413:3' Untranslated Regions; D004247:DNA / Q000235:genetics / Q000378:metabolism; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D016366:Open Reading Frames / Q000235:genetics; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D004247:DNA
keywords: DNA assembly; S. cerevisiae; synthetic biology; transcription unit; yeast Golden Gate