Difference between revisions of "Team:Wageningen UR/Basic Part"

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<h1><b>BioBricks overview</b></h1>
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<h1><b>Basic parts</b></h1>
  
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<p>Below, you can find all the biobricks we created!</p>
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<h2>Our best basic part: BBa_K19130025</h2>
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<br>
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<p>
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<a href="http://parts.igem.org/Part:BBa_K1913025">BBa_K1913025</a> consists of the natural FixK2 promoter with lac operons O1, O3
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</p>
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<p><br>
 +
<b>Below, you can find all basic parts we created!</b></p>
  
 
<table>
 
<table>
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     <td>chiB for Varroa destructor</td>
 
     <td>chiB for Varroa destructor</td>
 
     <td>Lisa</td>
 
     <td>Lisa</td>
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  </tr>
 
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  <tr>
 
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    <td><a href="http://parts.igem.org/Part:BBa_K1913002">BBa_K1913002</a></td>
 
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    <td>chiA device regulated by pBAD</td>
 
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    <td>Lisa</td>
 
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  </tr>
 
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  <tr>
 
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    <td><a href="http://parts.igem.org/Part:BBa_K1913003">BBa_K1913003</a></td>
 
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    <td>chiB device regulated by pBAD</td>
 
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    <td>Lisa</td>
 
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  </tr>
 
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  <tr>
 
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    <td><a href="http://parts.igem.org/Part:BBa_K1913005">BBa_K1913005</a></td>
 
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    <td>lux quorum sensing system + GFP reporter</td>
 
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    <td>Thomas</td>
 
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  </tr>
 
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  <tr>
 
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    <td><a href="http://parts.igem.org/Part:BBa_K1913006">BBa_K1913006</a></td>
 
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    <td>434- and lambda cI balance operon + mRFP reporter</td>
 
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    <td>Thomas</td>
 
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  </tr>
 
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  <tr>
 
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    <td><a href="http://parts.igem.org/Part:BBa_K1913007">BBa_K1913007</a></td>
 
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    <td>434- and lambda cI operon for tuning protein balance</td>
 
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    <td>Thomas</td>
 
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  </tr>
 
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  <tr>
 
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    <td><a href="http://parts.igem.org/Part:BBa_K19130014">BBa_K19130014</a></td>
 
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    <td>3-oxo-hexanoyl-HSL GFP reporter</td>
 
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    <td>Thomas</td>
 
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  </tr>
 
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  <tr>
 
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    <td><a href="http://parts.igem.org/Part:BBa_K19130016">BBa_K19130016</a></td>
 
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    <td>434- and lambda cI balance RFP reporter</td>
 
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    <td>Thomas</td>
 
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td><a href="http://parts.igem.org/Part:BBa_K19130010">BBa_K19130010</a></td>
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     <td><a href="http://parts.igem.org/Part:BBa_K1913019">BBa_K1913019</a></td>
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    <td>tetR QPI + mRFP</td>
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    <td>Carina</td>
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  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130011">BBa_K19130011</a></td>
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    <td>Vitamin b12 riboswitch + mRFP</td>
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    <td>Carina</td>
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  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130012">BBa_K19130012</a></td>
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    <td>Guanine riboswitch + guanine riboswitch</td>
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    <td>Carina</td>
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  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130019">BBa_K19130019</a></td>
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     <td>Guanine riboswitch BS-yxjA</td>
 
     <td>Guanine riboswitch BS-yxjA</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td><a href="http://parts.igem.org/Part:BBa_K19130020">BBa_K19130020</a></td>
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     <td><a href="http://parts.igem.org/Part:BBa_K1913020">BBa_K1913020</a></td>
 
     <td>mRFP with degredation tag</td>
 
     <td>mRFP with degredation tag</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td><a href="http://parts.igem.org/Part:BBa_K19130021">BBa_K19130021</a></td>
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     <td><a href="http://parts.igem.org/Part:BBa_K1913021">BBa_K1913021</a></td>
 
     <td>sGFP with defredation tag</td>
 
     <td>sGFP with defredation tag</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td><a href="http://parts.igem.org/Part:BBa_K19130022">BBa_K19130022</a></td>
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     <td><a href="http://parts.igem.org/Part:BBa_K1913022">BBa_K1913022</a></td>
 
     <td>Artificial FixK2 promoter with lac operon O1, O3 consitutive promoter </td>
 
     <td>Artificial FixK2 promoter with lac operon O1, O3 consitutive promoter </td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td><a href="http://parts.igem.org/Part:BBa_K19130023">BBa_K19130023</a></td>
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     <td><a href="http://parts.igem.org/Part:BBa_K1913023">BBa_K1913023</a></td>
 
     <td>Artificial FixK2 promoter with lac operon O1, O3 ompR</td>
 
     <td>Artificial FixK2 promoter with lac operon O1, O3 ompR</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td><a href="http://parts.igem.org/Part:BBa_K19130024">BBa_K19130024</a></td>
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     <td><a href="http://parts.igem.org/Part:BBa_K1913024">BBa_K1913024</a></td>
 
     <td>Artificial FixK2 promoter with two tetO operons</td>
 
     <td>Artificial FixK2 promoter with two tetO operons</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td><a href="http://parts.igem.org/Part:BBa_K19130025">BBa_K19130025</a></td>
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     <td><a href="http://parts.igem.org/Part:BBa_K1913025">BBa_K1913025</a></td>
 
     <td>Natural FixK2 promoter with lac operons O1, O3</td>
 
     <td>Natural FixK2 promoter with lac operons O1, O3</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td><a href="http://parts.igem.org/Part:BBa_K19130026">BBa_K19130026</a></td>
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     <td><a href="http://parts.igem.org/Part:BBa_K1913026">BBa_K1913026</a></td>
 
     <td>Natural FixK2 promoter with two tetO operons</td>
 
     <td>Natural FixK2 promoter with two tetO operons</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
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  <tr>
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    <tr>
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     <td><a href="http://parts.igem.org/Part:BBa_K19130027">BBa_K19130027</a></td>
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     <td><a href="http://parts.igem.org/Part:BBa_K1913013">BBa_K1913013</a></td>
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    <td>Wild type plac-FixK2 hybrid promoter with mRFP</td>
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    <td>Tianhe</td>
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  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130028">BBa_K19130028</a></td>
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    <td>Wild type ptet-FixK2 hybrid promoter with mRFP</td>
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    <td>Tianhe</td>
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  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130029">BBa_K19130029</a></td>
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    <td>Synthetic plac-FixK2 hybrid promoter +RBS with mRFP</td>
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    <td>Tianhe</td>
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  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130030">BBa_K19130030</a></td>
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    <td>Synthetic plac-FixK2 hybrid promoter+RBS with mRFP</td>
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    <td>Tianhe</td>
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−
  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130031">BBa_K19130031</a></td>
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    <td>Syntheitc ptet-FixK2 hybrid promoter+RBS with mRFP</td>
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    <td>Tianhe</td>
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  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130032">BBa_K19130032</a></td>
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    <td>Toggle Switch device</td>
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    <td>Tianhe</td>
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  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130033">BBa_K19130033</a></td>
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    <td>Toggle Switch device</td>
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    <td>Tianhe</td>
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  </tr>
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  <tr>
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    <td><a href="http://parts.igem.org/Part:BBa_K19130013">BBa_K19130013</a></td>
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     <td>Cry3Aa + RBS + TEV + His-tag</td>
 
     <td>Cry3Aa + RBS + TEV + His-tag</td>
 
     <td>Jaccoline/Linea</td>
 
     <td>Jaccoline/Linea</td>
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</table>
 
</table>
  
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<p>
 
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Besides, we submitted one part that cannot be classified as a biobrick because it has some illegal restriction sites:</p>
 
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<table> 
 
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  <tr>
 
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    <th>BioBrick number</th>
 
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    <th>BioBrick name</th>     
 
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    <th>Designer</th>
 
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  </tr>
 
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<tr>
 
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    <td><a href="http://parts.igem.org/Part:BBa_K19130015">BBa_K19130015</a></td>
 
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    <td>Cry3Aa with araC/pBAD</td>
 
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    <td>Jaccoline/Linea</td>
 
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  </tr>
 
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</table>
 
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<p>
 
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Initally, part BBa_K19130015 was made only to for testing the <a href="https://2016.igem.org/Team:Wageningen_UR/Description/Specificity#Assay"><i>in vitro</i> assay</a>, <b>jacco, please put the rest of your excuse here ;)</b>.
 
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</p>
 
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<p>
 
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<b>No Cas9 biobrick?!</b><br>
 
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When we started making the constructs for the <a href="https://2016.igem.org/Team:Wageningen_UR/Description/Biocontainment#SAA">Cas9 kill switch</a>, two approaches were taken: one was taking the Cas9 that is available in the iGEM registry (BBa_K1218011) as a starting point for making mutations and expressing Cas9. The other approach was starting with pdCas9 (Addgene plasmid # 46569). While cloning, it proved to be difficult to transfer BBa_K1218011 to another backbone (see also the <a href="https://2016.igem.org/Team:Wageningen_UR/Notebook/Cas9">notebook</a>). Furthermore, when cultures transformed with BBa_K1218011 were checked for Cas9 expression with SDS-PAGE, no convincing Cas9 band could be observed. Because of time limitations we decided to continue working with the Addgene construct for making the mutations, and chose an established system for protein expression. For that reason, no Cas9-biobricks were submitted. Furthermore, the pEVOL construct containing an aminoacyl-synthetase and a tRNA for introducing BipA in response to the TAG stopcodon were isolated from a strain kindly received from George Church (described in <a href="http://www.nature.com/nature/journal/v518/n7537/full/nature14121.html">Mandel <i>et al.</i>, 2015</a>). The MTA that was signed to receive the strain does not allow for redistribution. 
 
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</p>
 
  
  
 
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Revision as of 08:17, 17 October 2016

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Wageningen UR iGEM 2016

 

 

 

BioBricks

Basic parts

Our best basic part: BBa_K19130025


BBa_K1913025 consists of the natural FixK2 promoter with lac operons O1, O3


Below, you can find all basic parts we created!

BioBrick number BioBrick name Designer
BBa_K1913000 chiA for Varroa destructor Lisa
BBa_K1913001 chiB for Varroa destructor Lisa
BBa_K1913008 vitamin b12 riboswitch Carina
BBa_K1913009 Guanine riboswitch Carina
BBa_K1913019 Guanine riboswitch BS-yxjA Tianhe
BBa_K1913020 mRFP with degredation tag Tianhe
BBa_K1913021 sGFP with defredation tag Tianhe
BBa_K1913022 Artificial FixK2 promoter with lac operon O1, O3 consitutive promoter Tianhe
BBa_K1913023 Artificial FixK2 promoter with lac operon O1, O3 ompR Tianhe
BBa_K1913024 Artificial FixK2 promoter with two tetO operons Tianhe
BBa_K1913025 Natural FixK2 promoter with lac operons O1, O3 Tianhe
BBa_K1913026 Natural FixK2 promoter with two tetO operons Tianhe
BBa_K1913013 Cry3Aa + RBS + TEV + His-tag Jaccoline/Linea