Team:EPF Lausanne/Overview

From 2014.igem.org

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<h1 class="cntr">Project</h1>
<h1 class="cntr">Project</h1>
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<section id="title_intro">
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<h2 class="section-heading" id="title_intro">Introduction</h2>
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<h3 class="section-heading">DNA parts submitted by the 2014 EPFL iGEM team</h3>
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<p class="lead">
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Our team submitted a total of 55 Biobricks (biobrick 51 does not exist).</p>
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<p class="lead">
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In addition, 4 microfluidic designs have also been submitted to the registry.</p>
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<table class="table table-striped table-hover" id="biobricks_list">
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  <tr>
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    <th>Biobrick</th>
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    <th>What it is</th>
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    <th>Function</th>
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    <th>Why do we use it?</th>
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    <th>Group</th>
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  </tr>
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  <tr>
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    <td class="biobrick_name">BBa_K1486000</td>
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    <td>CpxR coding sequence</td>
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    <td>Transcription factor</td>
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    <td>To make most of our biobricks!</td>
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    <td>Bacteria</td>
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  </tr>
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  <tr>
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    <td class="biobrick_name">BBa_K1486001</td>
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    <td>CpxR under arabinose promoter</td>
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    <td>Treanscription factor</td>
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    <td> </td>
+
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    <td>Bacteria</td>
+
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  </tr>
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  <tr>
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    <td class="biobrick_name">BBa_K1486002</td>
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    <td>PAra + sfGFP CpxR [Nterm]</td>
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    <td>Expresses fused protein</td>
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    <td>Test CpxR expression & Ara promoter</td>
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    <td>Bacteria</td>
+
-
  </tr>
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  <tr>
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    <td class="biobrick_name">BBa_K1486003</td>
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    <td>Flexible linker</td>
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    <td>Attaches two proteins together</td>
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    <td> </td>
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    <td>Bacteria</td>
+
-
  </tr>
+
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  <tr>
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    <td class="biobrick_name">BBa_K1486004</td>
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    <td>Flexible linker</td>
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    <td>Attaches two proteins together</td>
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-
    <td> </td>
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    <td>Bacteria</td>
+
-
  </tr>
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  <tr>
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    <td class="biobrick_name">BBa_K1486005</td>
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    <td>PAra + CpxR sfGFP [Cterm]</td>
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    <td>Expresses fused protein</td>
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    <td>Test CpxR expression & Ara promoter</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
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  <tr>
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-
    <td class="biobrick_name">BBa_K1486006</td>
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    <td>IFP[1]</td>
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    <td>N terminus of split IFP</td>
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-
    <td> </td>
+
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    <td>Bacteria</td>
+
-
  </tr>
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  <tr>
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-
    <td class="biobrick_name">BBa_K1486007</td>
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    <td>IFP[2]</td>
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    <td>C terminus of split IFP</td>
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    <td> </td>
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    <td>Bacteria</td>
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-
  </tr>
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  <tr>
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    <td class="biobrick_name">BBa_K1486008</td>
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    <td>CxpR & Split IFP1.4 [Cterm + Cterm]</td>
+
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    <td>Two CpxR CDS, each C terminus attached to a moiety of IFP</td>
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    <td>Characterize CpxR dimerization</td>
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    <td>Bacteria</td>
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-
  </tr>
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  <tr>
+
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    <td class="biobrick_name">BBa_K1486009</td>
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    <td>CxpR & Split IFP1.4 [Nterm + Nterm]</td>
+
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    <td>Two CpxR CDS, each N terminus attached to a moiety of IFP</td>
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    <td>Characterize CpxR dimerization</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486010</td>
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-
    <td>CxpR & Split IFP1.4 [Nterm + Cterm]</td>
+
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    <td>Two CpxR CDS, each attached to a moiety of IFP</td>
+
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    <td>Characterize CpxR dimerization</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486011</td>
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-
    <td>CxpR & Split IFP1.4 [Cterm + Nterm]</td>
+
-
    <td>Two CpxR CDS, each attached to a moiety of IFP</td>
+
-
    <td>Characterize CpxR dimerization</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486012</td>
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    <td>CpxR + IFP[1]</td>
+
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    <td>CpxR with the Nterm moiety of IFP attached at its C terminus</td>
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    <td>Intermediate & control</td>
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-
    <td>Bacteria</td>
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-
  </tr>
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-
  <tr>
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    <td class="biobrick_name">BBa_K1486013</td>
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    <td>CpxR + IFP[2]</td>
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    <td>CpxR with the Cterm moiety of IFP attached at its C terminus</td>
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-
    <td>Intermediate & control</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
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-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486014</td>
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-
    <td>IFP[1] + CpxR</td>
+
-
    <td>CpxR with the Nterm moiety of IFP attached at its N terminus</td>
+
-
    <td>Intermediate & control</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486015</td>
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    <td>IFP[2] + CpxR</td>
+
-
    <td>CpxR with the Cterm moiety of IFP attached at its N terminus</td>
+
-
    <td>Intermediate & control</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
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    <td class="biobrick_name">BBa_K1486016</td>
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-
    <td>fLuc[1]</td>
+
-
    <td>N terminus moiety of the firefly luciferase</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
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    <td class="biobrick_name">BBa_K1486017</td>
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    <td>fLuc[2]</td>
+
-
    <td>C terminus moiety of the firefly luciferase</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486018</td>
+
-
    <td>PAra + fLuc[1] + fLuc[2]</td>
+
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    <td>Split firefly luciferase under arabinose promoter</td>
+
-
    <td>Control</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
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  <tr>
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    <td class="biobrick_name">BBa_K1486019</td>
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    <td>rLuc[1]</td>
+
-
    <td>C terminus moiety of the renilla luciferase</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
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    <td class="biobrick_name">BBa_K1486020</td>
+
-
    <td>rLuc[2]</td>
+
-
    <td>N terminus moiety of the renilla luciferase</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486021</td>
+
-
    <td>PAra + rLuc[1] + rLuc[2]</td>
+
-
    <td>Split renilla luciferase under arabinose promoter</td>
+
-
    <td>Control</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486022</td>
+
-
    <td>rLuc</td>
+
-
    <td>Full renilla luciferase</td>
+
-
    <td>Control</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
<tr>
+
-
    <td class="biobrick_name">BBa_K1486023</td>
+
-
    <td>Yeast sfGFP</td>
+
-
    <td>Superfolder GFP for yeast cells</td>
+
-
    <td>Reporter</td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486024</td>
+
-
    <td>Kan</td>
+
-
    <td>Yeast kanamycin resistance gene</td>
+
-
    <td>Selection marker</td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486025</td>
+
-
    <td>ADH1 terminator</td>
+
-
    <td>Terminator</td>
+
-
    <td> </td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486026</td>
+
-
    <td>Yeast sfGFP + Kan</td>
+
-
    <td>Yeast sfGFP attached to the yeast kanamycin resistance gene</td>
+
-
    <td>Control the expression of pbs2</td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
<tr>
+
-
    <td class="biobrick_name">BBa_K1486027</td>
+
-
    <td>rLuc + Kan</td>
+
-
    <td>Renilla luciferase attached to the kanamycin resistance gene</td>
+
-
    <td> </td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486028</td>
+
-
    <td>Yeast sfGFP[1]</td>
+
-
    <td>N terminal moiety of split yeast sfGFP</td>
+
-
    <td> </td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486029</td>
+
-
    <td>sfGFP[1] + kan</td>
+
-
    <td>Nterm moiety of split yeast sfGFP attached to yeast kanamycin resistance gene</td>
+
-
    <td> </td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486030</td>
+
-
    <td>rLuc[1] + kan</td>
+
-
    <td>Nterm moiety of split renilla luciferase attached to yeast kanamycin resistance gene</td>
+
-
    <td> </td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486031</td>
+
-
    <td>Ura</td>
+
-
    <td>CDS for Uracil (yeast selective purposes)</td>
+
-
    <td>Confer resistance to Uracil-deprived medium</td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
<tr>
+
-
    <td class="biobrick_name">BBa_K1486032</td>
+
-
    <td>Yeast sfGFP + Ura</td>
+
-
    <td>Yeast sfGFP attached to the Uracil CDS</td>
+
-
    <td>Control the expression of hog1</td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486033</td>
+
-
    <td>rLuc + Ura</td>
+
-
    <td>Renilla luciferase attached to the Uracil CDS</td>
+
-
    <td>Control the expression of hog1</td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486034</td>
+
-
    <td>yeast sfGFP[2]</td>
+
-
    <td>C terminal moiety of split the yeast sfGFP</td>
+
-
    <td> </td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486035</td>
+
-
    <td>yeast sfGFP[2] + Ura</td>
+
-
    <td>Cterm moiety of split yeast sfGFP attached to the Uracil CDS</td>
+
-
    <td> </td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
<tr>
+
-
    <td class="biobrick_name">BBa_K1486036</td>
+
-
    <td>rLuc[2] + Ura</td>
+
-
    <td>Cterm moiety of split renilla luciferase attached to the Uracil CDS</td>
+
-
    <td> </td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486037</td>
+
-
    <td>linker</td>
+
-
    <td>Attaches two proteins together</td>
+
-
    <td> </td>
+
-
    <td>Yeast</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486038</td>
+
-
    <td>sfGFP[1]</td>
+
-
    <td>N terminus moiety of split superfolder GFP</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486039</td>
+
-
    <td>sfGFP[2]</td>
+
-
    <td>C terminus moiety of split superfolder GFP</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486040</td>
+
-
    <td>sfGFP[1] + CpxR</td>
+
-
    <td>N terminus moiety of split sfGFP attached to CpxR</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486041</td>
+
-
    <td>sfGFP[2] + CpxR</td>
+
-
    <td>C terminus moiety of split sfGFP attached to CpxR</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486042</td>
+
-
    <td>LZip</td>
+
-
    <td>Monomer of leucine zipper TF</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486043</td>
+
-
    <td>LZip + split rLuc</td>
+
-
    <td>Two Leucine Zipper monomers, each attached to a different split rLuc moiety</td>
+
-
    <td>Control for split rLuc assays</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486044</td>
+
-
    <td>mut IFP[1]</td>
+
-
    <td>Biobrick-compatible IFP[1]</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486045</td>
+
-
    <td>mut IFP[2]</td>
+
-
    <td>Biobrick-compatible IFP[2]</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486046</td>
+
-
    <td>CpxR promoter FW</td>
+
-
    <td>CpxR binding-region in forward direction</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486047</td>
+
-
    <td>CpxR promoter RV</td>
+
-
    <td>CpxR binding-region in reverse direction</td>
+
-
    <td> </td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486048</td>
+
-
    <td>CpxR reporter</td>
+
-
    <td>Calgary's CpxR reporter repaired (sequence was missing)</td>
+
-
    <td>To see when CpxR is active</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486049</td>
+
-
    <td>CpxR promoter FW + RFP</td>
+
-
    <td>Reporter of CpxR</td>
+
-
    <td>Test the direction of the complete CpxR promoter</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486050</td>
+
-
    <td>CpxR promoter RV + RFP</td>
+
-
    <td>Reporter of CpxR</td>
+
-
    <td>Test the direction of the complete CpxR promoter</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486052</td>
+
-
    <td>Spacer</td>
+
-
    <td>40 bases placed between constructs</td>
+
-
    <td>Separate two constructs in the same plasmid</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486053</td>
+
-
    <td>Linker</td>
+
-
    <td>10 amino-acid linker</td>
+
-
    <td>Attach CheY/Z to split luciferases</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486054</td>
+
-
    <td>CheY/CheZ rLuc</td>
+
-
    <td>CheY and CheZ, each attached to a moiety of split renilla luciferase</td>
+
-
    <td>Positive control for the split rLuc</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486055</td>
+
-
    <td>CheY/CheZ fLuc</td>
+
-
    <td>CheY and CheZ, each attached to a moiety of split firefly luciferase</td>
+
-
    <td>Positive control for the split fLuc</td>
+
-
    <td>Bacteria</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td class="biobrick_name">BBa_K1486056</td>
+
-
    <td>CxpR & Split mut IFP1.4 [Cterm + Cterm]</td>
+
-
    <td>Two CpxR CDS, each C terminus attached to a moiety of the biobrick-compatible IFP</td>
+
-
    <td>Characterize CpxR dimerization</td>
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    <td>Bacteria</td>
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  </tr>
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</table>
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</section>
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<h2 class="section-heading" id="CpxPathway">Introduction</h2>
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<br /><br />
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<h2 class="section-heading" id="howitworks">Introduction</h2>
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<section id="CpxPathway">
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</div>
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<h3 class="section-heading">Microfluidics parts (chips created)</h3>
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<p class="lead">
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Our team designed and made 4 microfluidic chips. At the beginning, we also used the <a target="_blank" href="http://link.springer.com/protocol/10.1007%2F978-1-61779-292-2_6">MITOMI chip</a>.</p>
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<p class="lead">When designing the chips, the team took into account the future users and the current iGEM classification of parts. We considered it best to construct our chips as composite microfluidic parts, so their sub-parts could be used and combined in multiple ways. The flow and control layers can be separated and reused, as well as all the basic structures (chamber + connecting channel), nodes, array parts,...</p>
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<table class="table table-striped table-hover" id="chips_list">
 
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    <th>Name</th>
 
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    <th>Main Function</th>
 
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    <td>MITOMI modified</td>
 
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    <td>By using the MITOMI chip as a template, we designed our first chip that could squish the cells in the chamber.</td>
 
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  </tr>
 
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  <tr>
 
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    <td>SmashColi</td>
 
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    <td>To be able to separate the chip in 4 different compartments and apply 4 different pressures on each row of chambers.</td>
 
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  </tr>
 
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  <tr>
 
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    <td>BioPad</td>
 
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    <td>A large and simple microfluidic chip containing 6400 chambers in which the cells are contained in. Each chamber acts as a pixel for the BioPad project.</td>
 
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  </tr>
 
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  <tr>
 
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    <td>CleanColi</td>
 
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    <td>As a result of our Safety page, we decided to create a chip that is able to seal the bacteria in the chip, preventing them to leave the chip.</td>
 
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  </tr>
 
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  <tr>
 
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    <td>FilterColi</td>
 
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    <td>To successfully immerse cells in a certain solution, this chip was designed to flow in the new medium in the chambers instead of doing it by diffusion.</td>
 
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  </tr>
 
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</table>
 
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</section>
 
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</div>
 
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</div>

Revision as of 13:20, 15 October 2014

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