Difference between revisions of "Team:Wageningen UR/Parts"

Line 64: Line 64:
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130014">BBa_K19130014</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913014">BBa_K1913014</a></td>
 
     <td>3-oxo-hexanoyl-HSL GFP reporter</td>
 
     <td>3-oxo-hexanoyl-HSL GFP reporter</td>
 
     <td>Thomas</td>
 
     <td>Thomas</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130016">BBa_K19130016</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913016">BBa_K1913016</a></td>
 
     <td>434- and lambda cI balance RFP reporter</td>
 
     <td>434- and lambda cI balance RFP reporter</td>
 
     <td>Thomas</td>
 
     <td>Thomas</td>
Line 84: Line 84:
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130010">BBa_K19130010</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913010">BBa_K1913010</a></td>
 
     <td>tetR QPI + mRFP</td>
 
     <td>tetR QPI + mRFP</td>
 
     <td>Carina</td>
 
     <td>Carina</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130011">BBa_K19130011</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913011">BBa_K1913011</a></td>
 
     <td>Vitamin b12 riboswitch + mRFP</td>
 
     <td>Vitamin b12 riboswitch + mRFP</td>
 
     <td>Carina</td>
 
     <td>Carina</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130012">BBa_K19130012</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913012">BBa_K1913012</a></td>
 
     <td>Guanine riboswitch + guanine riboswitch</td>
 
     <td>Guanine riboswitch + guanine riboswitch</td>
 
     <td>Carina</td>
 
     <td>Carina</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130019">BBa_K19130019</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913019">BBa_K1913019</a></td>
 
     <td>Guanine riboswitch BS-yxjA</td>
 
     <td>Guanine riboswitch BS-yxjA</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130020">BBa_K19130020</a></td>
+
     <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>
     <td><a href="http://parts.igem.org/Part:BBa_K19130021">BBa_K19130021</a></td>
+
     <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>
     <td><a href="http://parts.igem.org/Part:BBa_K19130022">BBa_K19130022</a></td>
+
     <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>
     <td><a href="http://parts.igem.org/Part:BBa_K19130023">BBa_K19130023</a></td>
+
     <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>
     <td><a href="http://parts.igem.org/Part:BBa_K19130024">BBa_K19130024</a></td>
+
     <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>
     <td><a href="http://parts.igem.org/Part:BBa_K19130025">BBa_K19130025</a></td>
+
     <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>
     <td><a href="http://parts.igem.org/Part:BBa_K19130026">BBa_K19130026</a></td>
+
     <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>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130027">BBa_K19130027</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913027">BBa_K1913027</a></td>
 
     <td>Wild type plac-FixK2 hybrid promoter with mRFP</td>
 
     <td>Wild type plac-FixK2 hybrid promoter with mRFP</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130028">BBa_K19130028</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913028">BBa_K1913028</a></td>
 
     <td>Wild type ptet-FixK2 hybrid promoter with mRFP</td>
 
     <td>Wild type ptet-FixK2 hybrid promoter with mRFP</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130029">BBa_K19130029</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913029">BBa_K1913029</a></td>
 
     <td>Synthetic plac-FixK2 hybrid promoter +RBS with mRFP</td>
 
     <td>Synthetic plac-FixK2 hybrid promoter +RBS with mRFP</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130030">BBa_K19130030</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913030">BBa_K1913030</a></td>
 
     <td>Synthetic plac-FixK2 hybrid promoter+RBS with mRFP</td>
 
     <td>Synthetic plac-FixK2 hybrid promoter+RBS with mRFP</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130031">BBa_K19130031</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913031">BBa_K1913031</a></td>
 
     <td>Syntheitc ptet-FixK2 hybrid promoter+RBS with mRFP</td>
 
     <td>Syntheitc ptet-FixK2 hybrid promoter+RBS with mRFP</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130032">BBa_K19130032</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913032">BBa_K1913032</a></td>
 
     <td>Toggle Switch device</td>
 
     <td>Toggle Switch device</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130033">BBa_K19130033</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913033">BBa_K1913033</a></td>
 
     <td>Toggle Switch device</td>
 
     <td>Toggle Switch device</td>
 
     <td>Tianhe</td>
 
     <td>Tianhe</td>
Line 185: Line 185:
 
   </tr>
 
   </tr>
 
<tr>
 
<tr>
     <td><a href="http://parts.igem.org/Part:BBa_K19130015">BBa_K19130015</a></td>
+
     <td><a href="http://parts.igem.org/Part:BBa_K1913015">BBa_K1913015</a></td>
 
     <td>Cry3Aa with araC/pBAD</td>
 
     <td>Cry3Aa with araC/pBAD</td>
 
     <td>Jaccoline/Linea</td>
 
     <td>Jaccoline/Linea</td>
Line 192: Line 192:
  
 
<p>
 
<p>
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>.
+
Initally, part BBa_K1913015 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>.
 
</p>
 
</p>
  

Revision as of 08:20, 17 October 2016

Wageningen UR iGEM 2016

 

 

BioBricks overview

Below, you can find all the biobricks we created!

BioBrick number BioBrick name Designer
BBa_K1913000 chiA for Varroa destructor Lisa
BBa_K1913001 chiB for Varroa destructor Lisa
BBa_K1913002 chiA device regulated by pBAD Lisa
BBa_K1913003 chiB device regulated by pBAD Lisa
BBa_K1913005 lux quorum sensing system + GFP reporter Thomas
BBa_K1913006 434- and lambda cI balance operon + mRFP reporter Thomas
BBa_K1913007 434- and lambda cI operon for tuning protein balance Thomas
BBa_K1913014 3-oxo-hexanoyl-HSL GFP reporter Thomas
BBa_K1913016 434- and lambda cI balance RFP reporter Thomas
BBa_K1913008 vitamin b12 riboswitch Carina
BBa_K1913009 Guanine riboswitch Carina
BBa_K1913010 tetR QPI + mRFP Carina
BBa_K1913011 Vitamin b12 riboswitch + mRFP Carina
BBa_K1913012 Guanine riboswitch + 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_K1913027 Wild type plac-FixK2 hybrid promoter with mRFP Tianhe
BBa_K1913028 Wild type ptet-FixK2 hybrid promoter with mRFP Tianhe
BBa_K1913029 Synthetic plac-FixK2 hybrid promoter +RBS with mRFP Tianhe
BBa_K1913030 Synthetic plac-FixK2 hybrid promoter+RBS with mRFP Tianhe
BBa_K1913031 Syntheitc ptet-FixK2 hybrid promoter+RBS with mRFP Tianhe
BBa_K1913032 Toggle Switch device Tianhe
BBa_K1913033 Toggle Switch device Tianhe

Besides, we submitted one part that cannot be classified as a biobrick because it has some illegal restriction sites and it is in the pSB1A3 backbone:

BioBrick number BioBrick name Designer
BBa_K1913015 Cry3Aa with araC/pBAD Jaccoline/Linea

Initally, part BBa_K1913015 was made only to for testing the in vitro assay, jacco, please put the rest of your excuse here ;).

No Cas9 biobrick?!
When we started making the constructs for the Cas9 kill switch, 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 notebook). 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 Mandel et al., 2015). The MTA that was signed to receive the strain does not allow for redistribution.