BBF RFC 94: Type IIS Assembly for Bacterial Transcriptional Units: A Standardized Assembly Method for Building Bacterial Transcriptional Units Using the Type IIS Restriction Enzymes BsaI and BbsI
Name
BBFRFC94.pdf
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756.82 KB
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Author(s)
Haddock, Traci L.
Densmore, Douglas M.
Appleton, Evan
Carr, Swati
Iverson, Sonya
De Freitas, Monique
Jin, Shawn
Awtry, Jake
Desai, Devina
Lozanoski, Thomas
Date Issued
August 28, 2015
Series/Report no.
BBF RFC;94
Abstract
This RFC94 describes an assembly standard based on the Type IIS restriction enzymes BsaI and BbsI (also called BpiI). This assembly standard is based upon the Modular Cloning (MoClo) assembly strategy, which was introduced in 2011 by Weber et al. [1] and is based upon Golden Gate cloning [2].
In this RFC, we describe our proposed MoClo standard for generating a library of bacterial DNA parts for generating four-part transcriptional units (promoter : 5’UTR : CDS : 3’UTR). In this work, we define 5’UTRs as including ribosomal binding sites (RBS) and bi-cistronic design elements (BCDs) [3], and 3’UTRs as transcriptional terminators. The 2012-2014 BostonU iGEM teams completed this work and a more compact library has also been created based on this work [4].
Subjects
assembly standard
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Attribution 3.0 United States
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