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
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 th...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Technical Report |
| Language: | en_US |
| Published: |
2015
|
| Subjects: | |
| Online Access: | http://hdl.handle.net/1721.1/98267 |
| _version_ | 1826215397944197120 |
|---|---|
| author | Haddock, Traci L. Densmore, Douglas M. Appleton, Evan Carr, Swati Iverson, Sonya De Freitas, Monique Jin, Shawn Awtry, Jake Desai, Devina Lozanoski, Thomas Shah, Pooja Agarwal, Yash Lewis, Kathleen Pacheco, Alan |
| author_facet | Haddock, Traci L. Densmore, Douglas M. Appleton, Evan Carr, Swati Iverson, Sonya De Freitas, Monique Jin, Shawn Awtry, Jake Desai, Devina Lozanoski, Thomas Shah, Pooja Agarwal, Yash Lewis, Kathleen Pacheco, Alan |
| author_sort | Haddock, Traci L. |
| collection | MIT |
| description | 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]. |
| first_indexed | 2024-09-23T16:27:13Z |
| format | Technical Report |
| id | mit-1721.1/98267 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:27:13Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | mit-1721.1/982672019-04-10T09:32:07Z 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 Haddock, Traci L. Densmore, Douglas M. Appleton, Evan Carr, Swati Iverson, Sonya De Freitas, Monique Jin, Shawn Awtry, Jake Desai, Devina Lozanoski, Thomas Shah, Pooja Agarwal, Yash Lewis, Kathleen Pacheco, Alan assembly standard 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]. 2015-08-29T02:22:56Z 2015-08-29T02:22:56Z 2015-08-28 Technical Report http://hdl.handle.net/1721.1/98267 en_US BBF RFC;94 Attribution 3.0 United States http://creativecommons.org/licenses/by/3.0/us/ application/pdf |
| spellingShingle | assembly standard Haddock, Traci L. Densmore, Douglas M. Appleton, Evan Carr, Swati Iverson, Sonya De Freitas, Monique Jin, Shawn Awtry, Jake Desai, Devina Lozanoski, Thomas Shah, Pooja Agarwal, Yash Lewis, Kathleen Pacheco, Alan 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 |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | assembly standard |
| url | http://hdl.handle.net/1721.1/98267 |
| work_keys_str_mv | AT haddocktracil bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT densmoredouglasm bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT appletonevan bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT carrswati bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT iversonsonya bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT defreitasmonique bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT jinshawn bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT awtryjake bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT desaidevina bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT lozanoskithomas bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT shahpooja bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT agarwalyash bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT lewiskathleen bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi AT pachecoalan bbfrfc94typeiisassemblyforbacterialtranscriptionalunitsastandardizedassemblymethodforbuildingbacterialtranscriptionalunitsusingthetypeiisrestrictionenzymesbsaiandbbsi |