Team:Johns Hopkins-Wetware/Parts
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+ | <title>JHU iGEM 2012</title> | ||
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+ | <link rel="stylesheet" type="text/css" href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/templates/css/style?action=raw&ctype=text/css" /> | ||
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+ | <a href="https://2012.igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2012/0/00/Igem-logo-blue.png" width="100px"/></a> | ||
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+ | <div id="header_logo"> | ||
+ | <a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware"><img src="https://static.igem.org/mediawiki/2012/8/8f/Jhuigem2012header.png"/></a> | ||
+ | </div> <!--end div header_logo--> | ||
+ | <ul id="navbar"> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/Team">team</a> | ||
+ | </li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/Project">projects</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/Project">At a Glance</a></li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/etohproject">Ethanol control</a></li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/etohproject#modelanchor">Modeling</a></li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/lightproject">Optogenetic control</a></li> | ||
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+ | </ul> | ||
+ | </li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/etohnotebook">notebooks</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/etohnotebook">Ethanol control</a></li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/lightnotebook">Optogenetic control</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/yeastgoldengate">Yeast Golden Gate</a> | ||
+ | <ul> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/Parts">Parts</a></li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/yeastgoldengate">RFC88</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/humanpractice">human practice</a> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/Safety">safety</a> | ||
+ | </li> | ||
+ | <li><a href="https://2012.igem.org/Team:Johns_Hopkins-Wetware/requirements">Medal Fulfillment</a> | ||
+ | </li> | ||
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+ | <img src="yeast golden gate to BioBrick Conversion vectors" alt="Yeast Golden Gate to BioBrick Conversion Vectors"/> | ||
+ | </div> | ||
+ | <div class="content"> | ||
+ | <p> | ||
+ | We constructed and submitted 3 vectors to easily convert our Yeast Golden Gate Parts into BioBricks, one each for converting promoters, coding sequences, and terminators (BBa_K799024, BBa_K799025, BBa_K799024). These conversion vectors consist of the backbone standard BioBrick registry shipping vector pSB1C3 containing the standard BioBrick vector backbone part, which codes for RFP (BBa_J04450). The RFP is flanked by signature overhangs corresponding to a promoter, coding sequence, or terminator. In a one-pot digestion-ligation reaction, Yeast Golden Gate Parts can be converted to BioBricks through a red/white selection. | ||
+ | </p> | ||
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+ | We have sequenced these vectors across the RFP insert and signature overhangs to verify these parts, and have used them to successfully generate the BioBricks below. | ||
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+ | <img src="parts submitted using conversion vectors" alt="Parts Submitted using Conversion Vectors"/> | ||
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+ | <p> | ||
+ | We BioBricked 5 ethanol-induced yeast promoters with varying induction levels (BBa_K799001, BBa_K799003, BBa_K799005, BBa_K799007, BBa_K799009), the human cytochrome p450 2E1 (CYP2E1) used in our ethanol-control project (BBa_K799027), and the yeast terminator from the MFA2 gene (BBa_K799029). We also BioBricked a GFP sequence lacking BsaI and BsmBI sites for use with Yeast Golden Gate (BBa_K799028). We have sequence verified all of these parts. The ethanol-inducible promoters have been characterized through GFP expression under various ethanol levels. We have tested the ability of CYP2E1 to reduce ethanol concentration over a 24 hour fermentation. | ||
+ | </p> | ||
- | + | <groupparts>iGEM012 Johns_Hopkins-Wetware</groupparts> | |
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+ | <div class="content_header"> | ||
+ | <img src="yeast parts" alt="Yeast Parts generated from Parts Course"/> | ||
+ | </div> | ||
+ | <div class="content"> | ||
+ | <p> | ||
+ | In the first iteration of our BioParts course, we generated around 850 yeast parts including promoters, coding sequences, and terminators. These are parts that can be converted to the BioBrick standard using our conversion vectors above. | ||
+ | </p> | ||
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