Yeast Golden Gate: Standardized Assembly of S. cerevisiae Transcriptional Units
Name
BBFRFC88.pdf
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538.47 KB
Format
Adobe PDF
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Author(s)
Boeke, Jef
Mitchell, Leslie
Cai, Yizhi
Stracquadanio, Giovanni
Chuang, James
Tan, Scott
Heston, Margo
Wang, Jiarui
Kim, Dong Won
Noronha, Anne Marie
Date Issued
October 11, 2012
Series/Report no.
BBF RFC;88
Abstract
BBF RFC 88 describes a new standard for the assembly of basic Saccharomyces cerevisiae transcriptional units (TUs) consisting of a promoter/5’untranslated region (UTR), open reading frame (ORF), and 3’UTR/terminator. Note that we use the term “promoter” here to refer to both the promoter and the 5’ UTR, which we currently define as a single part. Future iterations of this standard will incorporate subdivision of currently defined parts e.g. into promoter and 5’ UTR. The standard makes use of the type IIS restriction enzyme BsaI to generate standardized and user‐defined ‘signature overhangs’, thus enabling directional and seamless TU assembly. RFC88 is supported by the Yeast Standardized Collection of Parts for Expression (SCoPE), a repository of subcloned and sequence verified parts compatible with this assembly standard. The Yeast SCoPE is currently populated by a large number of S. cerevisiae promoters and terminators that facilitate expression and characterization of non‐native ORFs.
Subjects
DNA assembly
yeast
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