Team:Edinburgh/BioBricks

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   <ul>
   <ul>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Project/Protocol">the protocol</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Project/Protocol">the protocol</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Project/BioBricks">submitted parts</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/BioBricks#Genomic">submitted parts</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Project/Results">results</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Results#Genomic">results</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Project/Future">future work</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Project/Future">the future</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Project/References">references</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Project/References">references</a></li>
   </ul>
   </ul>
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  <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial" class="dir">bacterial BRIDGEs</a>
  <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial" class="dir">bacterial BRIDGEs</a>
   <ul>
   <ul>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Core_repressilator">the repressilator</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Core_repressilator">the project</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_producer">red light</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_producer">red light</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_sensor">red sensor</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_sensor">red sensor</a></li>
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   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Green_light_producer">green light</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Green_light_producer">green light</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Green_light_sensor">green sensor</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Green_light_sensor">green sensor</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/BioBricks">submitted parts</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/BioBricks#Bacterial">submitted parts</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Results">results</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Results#Bacterial">results</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Future">future work</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Future">the future</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/References">references</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Bacterial/References">references</a></li>
   </ul>
   </ul>
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   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/Bacterial">the bacterial model</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/Bacterial">the bacterial model</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/Signalling">the signalling model</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/Signalling">the signalling model</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/Results">results</a></li>
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   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/Tools">tools</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/Future">future work</a></li>
+
  <li><a href="https://2010.igem.org/Team:Edinburgh/Results#Modelling">results</a></li>
 +
   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/Future">the future</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/References">references</a></li>
   <li><a href="https://2010.igem.org/Team:Edinburgh/Modelling/References">references</a></li>
   </ul>
   </ul>
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  <li><a href="https://2010.igem.org/Team:Edinburgh/Human" class="dir">human BRIDGEs</a>
  <li><a href="https://2010.igem.org/Team:Edinburgh/Human" class="dir">human BRIDGEs</a>
   <ul>
   <ul>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Human">human aspects</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Human/Communication">communication of science</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Human">results</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Human/Branding">iGEM survey</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Human">future work</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Human/Conversations">conversations</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Human">references</a></li>
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  <li><a href="https://2010.igem.org/Team:Edinburgh/Human/Epic">the epic</a></li>
 +
  <li><a href="https://2010.igem.org/Team:Edinburgh/Human/FutureApps">future applications</a></li>
 +
   <li><a href="https://2010.igem.org/Team:Edinburgh/Results#Human">further thoughts</a></li>
 +
  <li><a href="https://2010.igem.org/Team:Edinburgh/Human/References">references</a></li>
   </ul>
   </ul>
  </li>
  </li>
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  <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook" class="dir">lab notes&nbsp;&nbsp;&nbsp;</a>
  <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook" class="dir">lab notes&nbsp;&nbsp;&nbsp;</a>
   <ul>
   <ul>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook">collaboration</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Collaboration">collaboration</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook">BRIDGE</a></li>
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   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Attribution">attribution</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook">red light</a></li>
+
  <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/BRIDGE">BRIDGE</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook">red sensor</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Red_light_producer">red light</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook">blue light</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Red_light_sensor">red sensor</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook">blue sensor</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Blue_light_producer">blue light</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook">green light</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Blue_light_sensor">blue sensor</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook">green sensor</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Green_light_producer">green light</a></li>
-
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook">safety</a></li>
+
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Green_light_sensor">green sensor</a></li>
 +
   <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Modelling">modelling</a></li>
 +
  <li><a href="https://2010.igem.org/Team:Edinburgh/Notebook/Safety">safety</a></li>
   <li><a href="http://www.openwetware.org/wiki/French_Lab">protocols</a></li>
   <li><a href="http://www.openwetware.org/wiki/French_Lab">protocols</a></li>
   </ul>
   </ul>
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<br>
<br>
-
<p>For this part of the project we will have one main biobrick, which will be the construct required for the two-step markerless insertion, encompassing <i>cat</i> (chloramphenicol resistance) and <i>sacB</i> (prevents growth on sucrose).<br>
+
<p>For <a href="https://2010.igem.org/Team:Edinburgh/Project">the BRIDGE protocol</a> we have one main <a href="http://partsregistry.org/Part:BBa_K322922">BioBrick</a>, which is the construct <b>required</b> for the two-step markerless insertion, a composite construct of <i>cat</i> (<a href="http://partsregistry.org/Part:BBa_K322210">chloramphenicol resistance</a>) and <i>sacB</i> (<a href="http://partsregistry.org/Part:BBa_K322921">prevention of growth on sucrose</a>).</p>
-
Chloramphenicol resistance has already been well characterised in the registry, so our characterisation has mainly focused on <i>sacB</i>.
+
-
<br><br>
+
-
In addition we will probably be submitting the construct with the up- and down- stream sequences of useful genes which can be removed, e.g. tnaA, which produces indole - when removed <i>E. coli</i> no longer smells!</p>
+
<p>In addition, we <b>submitted</b> the <a href="http://partsregistry.org/Part:BBa_K322705">up-</a> and <a href="http://partsregistry.org/Part:BBa_K322706">downstream</a> sequences of an example of a useful gene to be removed from the <i>E. coli</i> genome using the BRIDGE protocol: <i>tnaA</i>, which produces indole, such that when it is removed <i>E. coli</i> no longer <b>smells</b>!</p>
<br>
<br>
 +
 +
<table border="0" cellspacing="10" cellpadding="5" style="background:transparent; border-color:transparent;">
 +
<tr>
 +
  <th width="100px"><u>Name</u></th>
 +
  <th width="100px"><u>Type</u></th>
 +
  <th width="100px"><u>Link</u></th>
 +
  <th><u>Description</u></th>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>cat</b></td>
 +
  <td>Translational Unit</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322210">BBa_K322210</a></td>
 +
  <td>Chloramphenicol acetyltransferase (chloramphenicol resistance gene).</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>sacB</b></td>
 +
  <td>Translational Unit</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322921">BBa_K322921</a></td>
 +
  <td><i>Bacillus subtilis</i> levansucrase, lethal to <i>E. coli</i> in the presence of sucrose.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>cat-sacB construct</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322922">BBa_K322922</a></td>
 +
  <td>Construct necessary for BRIDGE protocol.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>tnaA upstream region</b></td>
 +
  <td>Other</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322705">BBa_K322705</a></td>
 +
  <td>Upstream region of <i>E. coli</i> tryptophanase locus, used for targetting genes to this locus using BRIDGE.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>tnaA downstream region</b></td>
 +
  <td>Other</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322706">BBa_K322706</a></td>
 +
  <td>Downstream region of <i>E. coli</i> tryptophanase locus, used for targetting genes to this locus using BRIDGE.</td>
 +
</tr>
 +
</table>
 +
<br>
<br>
 +
<br>
 +
 +
<a name="Bacterial" id="Bacterial"></a><h2>BioBricks: Bacterial BRIDGEs</h2>
 +
<br>
 +
 +
<p>The light-sensing and light-production BioBricks of various emission / absorbance wavelengths developed in the <a href="https://2010.igem.org/Team:Edinburgh/Bacterial">FORTH light communication framework</a> are listed below. A few are <b>resubmissions</b> of existing parts (noted as such in their descriptions), whilst others are composite devices with promoters and reporter systems.</p>
 +
 +
<p>The BioBricks for <a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_producer">red light production</a> are two variants of mutated firefly luciferase (<a href="http://partsregistry.org/Part:BBa_K322246">S284T</a> and <a href="http://partsregistry.org/Part:BBa_K322211">356K</a>) and their composite reporter systems.</p>
 +
<br>
 +
 +
<table border="0" cellspacing="10" cellpadding="5" style="background:transparent; border-color:transparent;">
 +
<tr>
 +
  <th width="100px"><u>Name</u></th>
 +
  <th width="100px"><u>Type</u></th>
 +
  <th width="100px"><u>Link</u></th>
 +
  <th><u>Description</u></th>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>Firefly luciferase S284T mutant</b></td>
 +
  <td>Coding</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322246">BBa_K322246</a></td>
 +
  <td>Firefly luciferase from <i>Photinus pyralis</i> mutated towards the red spectrum.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>S284T mutant luciferase under lac promoter</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322247">BBa_K322247</a></td>
 +
  <td>Lac promoter followed by firefly luciferase S284T mutant.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>Firefly luciferase 356K mutant</b></td>
 +
  <td>Coding</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322211">BBa_K322211</a></td>
 +
  <td>Firefly luciferase S284T mutant from <i>Photinus pyralis</i> re-mutated towards the red spectrum.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>356K mutant luciferase under lac promoter</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322212">BBa_K322212</a></td>
 +
  <td>Lac promoter followed by firefly luciferase 356K mutant.</td>
 +
</tr>
 +
</table>
 +
 +
<br><br>
 +
<p>The BioBricks for <a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Blue_light_producer">blue light production</a> are corrected variants of the <a href="http://partsregistry.org/Part:BBa_K322139">LuxAB</a>, <a href="http://partsregistry.org/Part:BBa_K322312">LuxCDE</a>, and <a href="http://partsregistry.org/Part:BBa_K322007">LumP</a> proteins from <a href="https://2009.igem.org/Team:Edinburgh">Edinburgh 2009</a> in various combinations, contained in new plasmids and with various reporter systems.</p>
 +
<br>
 +
 +
<table border="0" cellspacing="10" cellpadding="5" style="background:transparent; border-color:transparent;">
 +
<tr>
 +
  <th width="100px"><u>Name</u></th>
 +
  <th width="100px"><u>Type</u></th>
 +
  <th width="100px"><u>Link</u></th>
 +
  <th><u>Description</u></th>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>luxAB</b></td>
 +
  <td>Translational Unit</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322139">BBa_K322139</a></td>
 +
  <td>Bacterial luciferase producing blue light (from <i>X. luminescens</i>).</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>luxCDE</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322312">BBa_K322312</a></td>
 +
  <td>Codes for a fatty acid reductase complex that makes the fatty acids necessary for LuxAB.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>lumP</b></td>
 +
  <td>Translational Unit</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322007">BBa_K322007</a></td>
 +
  <td>Lumazine protein to shift the wavelength of LuxAB to blue.</td>
 +
</tr>
 +
</table>
</div>
</div>
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<br>
<br>
-
<a name="Bacterial" id="Bacterial"></a><h2>BioBricks: Bacterial BRIDGEs</h2>
+
<table border="0" cellspacing="10" cellpadding="5" style="background:transparent; border-color:transparent;">
 +
<tr>
 +
  <th width="100px"><u>Name</u></th>
 +
  <th width="100px"><u>Type</u></th>
 +
  <th width="100px"><u>Link</u></th>
 +
  <th><u>Description</u></th>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>luxAB under lac promoter</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322140">BBa_K322140</a></td>
 +
  <td>Lac promoter followed by luxAB.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>luxAB and luxCDE under lac promoter</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322141">BBa_K322141</a></td>
 +
  <td>Lac promoter followed by luxAB and luxCDE.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>luxAB and lumP</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322149">BBa_K322149</a></td>
 +
  <td>luxAB and lumP composite part.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>luxAB and lumP under lac promoter</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322150">BBa_K322150</a></td>
 +
  <td>Lac promoter followed by luxAB and lumP</td>
 +
</tr>
 +
</table>
 +
 
 +
<br><br>
 +
<p>The BioBricks for <a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Green_light_producer">green light production</a> are a modified version of the <a href="http://partsregistry.org/Part:BBa_K322237">wildtype firefly luciferase</a> from <a href="https://2007.igem.org/Ljubljana">Ljubljana 2007</a> and a <a href="http://partsregistry.org/Part:BBa_K322451">codon-optimised variant</a> with brighter output.</p>
<br>
<br>
-
<p><b>Biobrick #1</b>
+
<table border="0" cellspacing="10" cellpadding="5" style="background:transparent; border-color:transparent;">
 +
<tr>
 +
  <th width="100px"><u>Name</u></th>
 +
  <th width="100px"><u>Type</u></th>
 +
  <th width="100px"><u>Link</u></th>
 +
  <th><u>Description</u></th>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>Firefly luciferase</b></td>
 +
  <td>Coding</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322237">BBa_K322237</a></td>
 +
  <td>Ljubljana 2007's <a href="http://partsregistry.org/Part:BBa_I712019">BBa_I712019</a> in pSB1C3, from <i>Photinus pyralis</i>.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>Wildtype luciferase under lac promoter</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322238">BBa_K322238</a></td>
 +
  <td>Lac promoter followed by wildtype luciferase.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>Codon optimised bright firefly luciferase</b></td>
 +
  <td>Translational Unit</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322451">BBa_K322451</a></td>
 +
  <td>Firefly luciferase from <i>Photinus pyralis</i> mutated for increased bioluminescence.</td>
 +
</tr>
 +
</table>
<br><br>
<br><br>
-
A brighter luciferase: we had synthesised a version of the Photinus pyralis (Ppy) luciferase with 3 specific amino acid mutations: Ile423Leu Asp436Gly and Leu530Arg. These mutations increase the light intensity of the luciferase by a factor of 12.5.<br><br>
+
<p>The BioBricks for <a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_sensor">red light sensing</a> are the updated <a href="http://partsregistry.org/Part:BBa_K322124">Cph8</a> systems from <a href="http://partsregistry.org/Coliroid">UT Austin 2004</a> and <a href="https://2008.igem.org/Team:Harvard">Harvard 2008</a> in a variety of combinations, contained in a new plasmid and with two different reporter systems.</p>
 +
<br>
-
<a href="http://partsregistry.org/Part:BBa_K322451">BBa_K322451</a>
+
<table border="0" cellspacing="10" cellpadding="5" style="background:transparent; border-color:transparent;">
-
<br><br>
+
<tr>
 +
  <th width="100px"><u>Name</u></th>
 +
  <th width="100px"><u>Type</u></th>
 +
  <th width="100px"><u>Link</u></th>
 +
  <th><u>Description</u></th>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>Phycocyanobilin synthesis operon</b></td>
 +
  <td>Translational Unit</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322122">BBa_K322122</a></td>
 +
  <td>Harvard 2008's <a href="http://partsregistry.org/Part:BBa_K098010">BBa_K098010</a> in pSB1C3.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>Phycocyanobilin synthesis operon without terminator</b></td>
 +
  <td>Translational Unit</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322123">BBa_K322123</a></td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K098010">BBa_K098010</a> without terminator, allowing addition of cph8.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>cph8 with RBS</b></td>
 +
  <td>Translational Unit</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322124">BBa_K322124</a></td>
 +
  <td>Light sensing protein for red light (<a href="http://partsregistry.org/Part:BBa_I15010">BBa_I15010</a> in pSB1C3).</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>cph8 and reporter system (lacZ)</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322125">BBa_K322125</a></td>
 +
  <td>Red light sensor with lacZ reporter system.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>cph8 and reporter system (EYFP)</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322126">BBa_K322126</a></td>
 +
  <td>Red light sensor with EYFP reporter system.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>Phycocyanobilin synthesis genes with cph8</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322127">BBa_K322127</a></td>
 +
  <td>The entire phycocyanobilin - cph8 system, without reporter system.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>Phycocyanobilin synthesis genes with cph8 and EYFP reporter system</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322128">BBa_K322128</a></td>
 +
  <td>The entire phycocyanobilin - cph8 system, with EYFP reporter system.</td>
 +
</tr>
 +
</table>
-
Currently, this part is only present in sequence form. Physical DNA will be submitted shortly.  
+
</div>
 +
 
 +
<div id="body2" style="padding: 0px 60px 10px 60px; height: 998px">
 +
<br>
 +
<br>
 +
 
 +
<p>The BioBrick for <a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Blue_light_sensor">blue light sensing</a> is a <a href="http://partsregistry.org/Part:BBa_K322999">LovTAP reporter system</a> based on work by <a href="https://2009.igem.org/Team:EPF-Lausanne">Lausanne 2009</a>.</p>
 +
<br>
 +
 
 +
<table border="0" cellspacing="10" cellpadding="5" style="background:transparent; border-color:transparent;">
 +
<tr>
 +
  <th width="100px"><u>Name</u></th>
 +
  <th width="100px"><u>Type</u></th>
 +
  <th width="100px"><u>Link</u></th>
 +
  <th><u>Description</u></th>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>LovTAP and reporter system (RFP)</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322999">BBa_K322999</a></td>
 +
  <td>Blue light sensor with RFP reporter system.</td>
 +
</tr>
 +
</table>
<br><br>
<br><br>
-
Reference: Fujii H, Noda K, Asami Y, Kuroda A, Sakata M, Tokida A (2007) Increase in bioluminescence intensity of firefly luciferase using genetic modification. ''Analytical Biochemistry'' '''366''': 131-136 </p>
+
<p>The BioBricks for the <a href="https://2010.igem.org/Team:Edinburgh/Bacterial/Green_light_sensor">green light sensing</a> are the <a href="http://partsregistry.org/Part:BBa_K322119">CcaS-PhoR light sensor</a> and the associated parts necessary to create the desired reporter system.</p>
 +
<br>
 +
 
 +
<table border="0" cellspacing="10" cellpadding="5" style="background:transparent; border-color:transparent;">
 +
<tr>
 +
  <th width="100px"><u>Name</u></th>
 +
  <th width="100px"><u>Type</u></th>
 +
  <th width="100px"><u>Link</u></th>
 +
  <th><u>Description</u></th>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>phoA promoter</b></td>
 +
  <td>Regulatory</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322115">BBa_K322115</a></td>
 +
  <td>Promoter activated by the CcaS-PhoR pathway.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>phoA promoter with lacZ</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322117">BBa_K322117</a></td>
 +
  <td>phoA promoter coupled to lacZ reporter.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>CcaS-PhoR</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322119">BBa_K322119</a></td>
 +
  <td>Green light sensor.</td>
 +
</tr>
 +
<tr>
 +
  <td align="left"><b>CcaS-PhoR and reporter system (lacZ)</b></td>
 +
  <td>Composite</td>
 +
  <td><a href="http://partsregistry.org/Part:BBa_K322120">BBa_K322120</a></td>
 +
  <td>Green light sensor with <i>lacZ</i> reporter system.</td>
 +
</tr>
 +
</table>
<br>
<br>
Line 159: Line 445:
<center><a href="#top" class="dir"><img width="100" src="https://static.igem.org/mediawiki/2010/9/9f/Ed10-RTT.png"></a></center>
<center><a href="#top" class="dir"><img width="100" src="https://static.igem.org/mediawiki/2010/9/9f/Ed10-RTT.png"></a></center>
 +
</div>
 +
 +
<div id="windowbox" style="border: .2em solid #660000; padding: 5px; position:fixed; top:50%; right:30px; width:8%;">
 +
<span style="color:ivory;">Throughout this wiki there are words in <b>bold</b> that indicate a relevance to <b>human aspects</b>. It will become obvious that <b>human aspects</b> are a part of almost everything in <b>iGEM</b>.</span>
 +
</div>
</div>

Latest revision as of 02:16, 28 October 2010







BioBricks: Genomic BRIDGEs


For the BRIDGE protocol we have one main BioBrick, which is the construct required for the two-step markerless insertion, a composite construct of cat (chloramphenicol resistance) and sacB (prevention of growth on sucrose).

In addition, we submitted the up- and downstream sequences of an example of a useful gene to be removed from the E. coli genome using the BRIDGE protocol: tnaA, which produces indole, such that when it is removed E. coli no longer smells!


Name Type Link Description
cat Translational Unit BBa_K322210 Chloramphenicol acetyltransferase (chloramphenicol resistance gene).
sacB Translational Unit BBa_K322921 Bacillus subtilis levansucrase, lethal to E. coli in the presence of sucrose.
cat-sacB construct Composite BBa_K322922 Construct necessary for BRIDGE protocol.
tnaA upstream region Other BBa_K322705 Upstream region of E. coli tryptophanase locus, used for targetting genes to this locus using BRIDGE.
tnaA downstream region Other BBa_K322706 Downstream region of E. coli tryptophanase locus, used for targetting genes to this locus using BRIDGE.


BioBricks: Bacterial BRIDGEs


The light-sensing and light-production BioBricks of various emission / absorbance wavelengths developed in the FORTH light communication framework are listed below. A few are resubmissions of existing parts (noted as such in their descriptions), whilst others are composite devices with promoters and reporter systems.

The BioBricks for red light production are two variants of mutated firefly luciferase (S284T and 356K) and their composite reporter systems.


Name Type Link Description
Firefly luciferase S284T mutant Coding BBa_K322246 Firefly luciferase from Photinus pyralis mutated towards the red spectrum.
S284T mutant luciferase under lac promoter Composite BBa_K322247 Lac promoter followed by firefly luciferase S284T mutant.
Firefly luciferase 356K mutant Coding BBa_K322211 Firefly luciferase S284T mutant from Photinus pyralis re-mutated towards the red spectrum.
356K mutant luciferase under lac promoter Composite BBa_K322212 Lac promoter followed by firefly luciferase 356K mutant.


The BioBricks for blue light production are corrected variants of the LuxAB, LuxCDE, and LumP proteins from Edinburgh 2009 in various combinations, contained in new plasmids and with various reporter systems.


Name Type Link Description
luxAB Translational Unit BBa_K322139 Bacterial luciferase producing blue light (from X. luminescens).
luxCDE Composite BBa_K322312 Codes for a fatty acid reductase complex that makes the fatty acids necessary for LuxAB.
lumP Translational Unit BBa_K322007 Lumazine protein to shift the wavelength of LuxAB to blue.


Name Type Link Description
luxAB under lac promoter Composite BBa_K322140 Lac promoter followed by luxAB.
luxAB and luxCDE under lac promoter Composite BBa_K322141 Lac promoter followed by luxAB and luxCDE.
luxAB and lumP Composite BBa_K322149 luxAB and lumP composite part.
luxAB and lumP under lac promoter Composite BBa_K322150 Lac promoter followed by luxAB and lumP


The BioBricks for green light production are a modified version of the wildtype firefly luciferase from Ljubljana 2007 and a codon-optimised variant with brighter output.


Name Type Link Description
Firefly luciferase Coding BBa_K322237 Ljubljana 2007's BBa_I712019 in pSB1C3, from Photinus pyralis.
Wildtype luciferase under lac promoter Composite BBa_K322238 Lac promoter followed by wildtype luciferase.
Codon optimised bright firefly luciferase Translational Unit BBa_K322451 Firefly luciferase from Photinus pyralis mutated for increased bioluminescence.


The BioBricks for red light sensing are the updated Cph8 systems from UT Austin 2004 and Harvard 2008 in a variety of combinations, contained in a new plasmid and with two different reporter systems.


Name Type Link Description
Phycocyanobilin synthesis operon Translational Unit BBa_K322122 Harvard 2008's BBa_K098010 in pSB1C3.
Phycocyanobilin synthesis operon without terminator Translational Unit BBa_K322123 BBa_K098010 without terminator, allowing addition of cph8.
cph8 with RBS Translational Unit BBa_K322124 Light sensing protein for red light (BBa_I15010 in pSB1C3).
cph8 and reporter system (lacZ) Composite BBa_K322125 Red light sensor with lacZ reporter system.
cph8 and reporter system (EYFP) Composite BBa_K322126 Red light sensor with EYFP reporter system.
Phycocyanobilin synthesis genes with cph8 Composite BBa_K322127 The entire phycocyanobilin - cph8 system, without reporter system.
Phycocyanobilin synthesis genes with cph8 and EYFP reporter system Composite BBa_K322128 The entire phycocyanobilin - cph8 system, with EYFP reporter system.


The BioBrick for blue light sensing is a LovTAP reporter system based on work by Lausanne 2009.


Name Type Link Description
LovTAP and reporter system (RFP) Composite BBa_K322999 Blue light sensor with RFP reporter system.


The BioBricks for the green light sensing are the CcaS-PhoR light sensor and the associated parts necessary to create the desired reporter system.


Name Type Link Description
phoA promoter Regulatory BBa_K322115 Promoter activated by the CcaS-PhoR pathway.
phoA promoter with lacZ Composite BBa_K322117 phoA promoter coupled to lacZ reporter.
CcaS-PhoR Composite BBa_K322119 Green light sensor.
CcaS-PhoR and reporter system (lacZ) Composite BBa_K322120 Green light sensor with lacZ reporter system.



Throughout this wiki there are words in bold that indicate a relevance to human aspects. It will become obvious that human aspects are a part of almost everything in iGEM.