Difference between revisions of "Team:Marburg/Parts"

 
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{{Marburg}}
 
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<div class="titleWrapper"><div class="titleBackground" style="background-image:none"></div><div class="title">Parts Overview</div></div>
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<div class="titleWrapper"><div class="titleBackground" style="background-image:url(https://static.igem.org/mediawiki/2018/2/27/T--Marburg--home_3-2.jpeg)"></div><div class="title">Parts Overview</div></div>
 
<article>
 
<article>
 
 
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<div class="content">
 
<div class="content">
As an extension of our basic part <i>tfoX</i> we created a full transcriptional unit enabling you to perform natural transformation in <i>V. natriegens</i>.
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<figure style="width:50%; float:left; margin:0.5em;"><img style="display:block; margin:0 auto 0 auto; width:100%;" src="https://static.igem.org/mediawiki/2018/3/3e/T--Marburg--tfoxsequencing.svg" alt="Relative promoter strength of Anderson promoters">
First we started assembling a  full transcriptional unit using a constitutive promoter. Therefore, we assembled <i>tfoX</i> together with the constitutive promoters J23100 or J23110 into a transcriptional unit by golden gate assembly. After transformation of this reactions into <i>E. coli</i> we were able to get colonies that seemed to be positive for the Lvl1 plasmids encoding <i>tfoX</i> because they showed the right size of fragment after plasmid preparation and test digestion, whenever we sent them for sequencing, some basepairs of the <i>tfox</i> gene were always missing (Figure 1).
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<figcaption><b>Figure 1:</b>Alignment of sequencing results of our first constructed Lvl1 <i>tfoX</i> plasmids showing missing basepairs at places at which black fragments are missing.</figcaption>
 +
            </figure>
 +
 
 +
<p>As an extension of our basic part <i>tfoX</i> we created a full transcriptional unit enabling you to perform natural transformation in <i>V. natriegens</i>.
 +
First we started assembling a  full transcriptional unit using a constitutive promoter. Therefore, we assembled <i>tfoX</i> together with the constitutive promoters J23100 or J23110 into a transcriptional unit by golden gate assembly. After transformation of this reactions into <i>E. coli</i> we were able to get colonies that seemed to be positive for the Lvl1 plasmids encoding <i>tfoX</i> because they showed the right size of fragment after plasmid preparation and test digestion, whenever we sent them for sequencing, some basepairs of the <i>tfox</i> gene were always missing (Figure 1).</p>
  
<figure style="width: 40%; float:left; margin:0.5em;"><img style="display:block; margin:0 auto 0 auto; width:100%;" src="https://static.igem.org/mediawiki/2018/3/3e/T--Marburg--tfoxsequencing.svg" alt="Relative promoter strength of Anderson promoters">
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<figure style="width: 50%; float: right; margin: 0.5em;"><img style="display:block; margin:0 auto 0 auto; width:100%;" src="https://static.igem.org/mediawiki/2018/4/4d/T--Marburg--pTet.png" alt="Alignment of sequencing results of our first constructed Lvl1 <i>tfoX</i> plasmids showing missing basepairs.">
 
<figcaption><b>Figure 2:</b> Relative promoter strength of Anderson promoters  
 
<figcaption><b>Figure 2:</b> Relative promoter strength of Anderson promoters  
 
Data were normalized over the strongest construct J23100. Error bars represent the standard deviation of the measurements of three independent experiments</figcaption>
 
Data were normalized over the strongest construct J23100. Error bars represent the standard deviation of the measurements of three independent experiments</figcaption>
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The pTet promoter can be induced by the tetracycline derivative anhydrotetracycline (ATc). ATc is much less cytotoxic than IPTG but still capable of binding and altering the structure of the repressor TetR, leading to release of the promoter and enabling transcription.
 
The pTet promoter can be induced by the tetracycline derivative anhydrotetracycline (ATc). ATc is much less cytotoxic than IPTG but still capable of binding and altering the structure of the repressor TetR, leading to release of the promoter and enabling transcription.
  
 
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<figure style="width: 25%; float:left; margin: 0.5em;"><img style="width: 100%;" src="https://static.igem.org/mediawiki/2018/6/64/T--Marburg--Testdigesttfoxlevel1.svg" alt="1 %- Agarose gel of the test digest.">
<figure style="width: 30%; float: right; margin: 0.5em;"><img style="display:block; margin:0 auto 0 auto; width:100%;" src="https://static.igem.org/mediawiki/2018/4/4d/T--Marburg--pTet.png" alt="Alignment of sequencing results of our first constructed Lvl1 <i>tfoX</i> plasmids showing missing basepairs.">
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<figcaption><b>Figure 1:</b>Alignment of sequencing results of our first constructed Lvl1 <i>tfoX</i> plasmids showing missing basepairs at places at which black fragments are missing.</figcaption>
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            </figure>
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<p>Using this suitable promoter for <i>V. natriegens</i> we successfully assembled a plasmid containing a full transcriptional unit with a pTet promoter and did not had any problems during the cloning process. By Test Digestion and sequencing we could prove, that now the sequence of Tfox was correct (Figure 3 and 4).</p>
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As we have already shown in the chapter of our basic part, we were able to perform natural transformation using this plasmid and could demonstrate, that GFP which is encoded on pYTK is expressed by <i>V. natriegens</i>.
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<figure style="width: 50%; margin: 0.5em;"><img style="width: 100%;" src="https://static.igem.org/mediawiki/2018/6/64/T--Marburg--Testdigesttfoxlevel1.svg" alt="1 %- Agarose gel of the test digest.">
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<figcaption><b>Figure 3:</b>1 %- Agarose gel of the test digest. 2-logDNA ladder is used as marker on agarose gels.</figcaption>
 
<figcaption><b>Figure 3:</b>1 %- Agarose gel of the test digest. 2-logDNA ladder is used as marker on agarose gels.</figcaption>
 
             </figure>
 
             </figure>
  
  <figure style="width: 50%; margin: 0.5em;"><img style="width: 100%;" src="https://static.igem.org/mediawiki/2018/a/a8/T--Marburg--sequencingtfox.png" alt="Alignment of sequencing results of our final constructed Lvl1 <i>tfoX</i> plasmid showing the correct sequence.">
+
<p>Using this suitable promoter for <i>V. natriegens</i> we successfully assembled a plasmid containing a full transcriptional unit with a pTet promoter and did not had any problems during the cloning process. By Test Digestion and sequencing we could prove, that now the sequence of Tfox was correct (Figure 3 and 4).</p>
 +
<figure style="width: 50%; margin: 0.5em; float:right;"><img style="width: 100%;" src="https://static.igem.org/mediawiki/2018/a/a8/T--Marburg--sequencingtfox.png" alt="Alignment of sequencing results of our final constructed Lvl1 <i>tfoX</i> plasmid showing the correct sequence.">
 
<figcaption><b>Figure 4:</b>Alignment of sequencing results of our final constructed Lvl1 <i>tfoX</i> plasmid showing the correct sequence.</figcaption>
 
<figcaption><b>Figure 4:</b>Alignment of sequencing results of our final constructed Lvl1 <i>tfoX</i> plasmid showing the correct sequence.</figcaption>
 
             </figure>
 
             </figure>
 +
As we have already shown in the chapter of our basic part, we were able to perform natural transformation using this plasmid and could demonstrate, that GFP which is encoded on pYTK is expressed by <i>V. natriegens</i>.
 +
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 +
 
  
 
<figure style=" width: 50%; margin: 0.5em;"><img style="width: 100%;" src="https://static.igem.org/mediawiki/2018/e/ed/T--Marburg--pYTKtransformation.svg" alt="Plates showing successful transformation of pYTK.">
 
<figure style=" width: 50%; margin: 0.5em;"><img style="width: 100%;" src="https://static.igem.org/mediawiki/2018/e/ed/T--Marburg--pYTKtransformation.svg" alt="Plates showing successful transformation of pYTK.">
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</ul>
 
</ul>
 
</div>  
 
</div>  
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</div>
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<div class="collapsible">
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<div class="btn_expand">
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<div class="btn_e_title">Marburg Collection</div>
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<div class="btn_e_desc">
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We created a novel golden-gate based cloning toolbox consisting of 123 parts. Plasmid maps of all parts can be downloaded here.
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</div>
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</div>
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<div class="content">
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<h3> Parts of the Marburg Collection </h3>
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 +
<div>  <a href=https://static.igem.org/mediawiki/parts/6/64/T--Marburg--Marburg_Collection.zip>Download the "Marburg Collection"</abbr></a>
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</div>
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<div>
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 +
<form action="https://static.igem.org/mediawiki/parts/6/64/T--Marburg--Marburg_Collection.zip" method="get" target="_blank">
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<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
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</form>
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</div>
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<div class="collapsible">
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<div class="btn_expand">5'Connector</div>
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<div class="content">
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<div>  <a href=https://static.igem.org/mediawiki/parts/d/d0/T--Marburg--5%27Connectors.zip>Download all 5'Connectors</abbr></a>
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</div>
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<div>
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<form action="https://static.igem.org/mediawiki/parts/d/d0/T--Marburg--5%27Connectors.zip" method="get" target="_blank">
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<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
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</form>
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</div>
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                <table>
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                <thead>
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                    <tr>
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                        <th> BioBrick</th>
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                        <th> Name</th>
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                        <th> Plasmid</th>
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                    </tr>
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                </thead>
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                <tbody>
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 +
<tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560011">K2560011 </a> </td>
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                        <td>5'Connector Dummy</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                                               
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<tr>
 +
                    </tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560055">K2560055 </a> </td>
 +
                        <td>1-6 Connector</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
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<tr>
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                        <td> <a href="http://parts.igem.org/Part:BBa_K2560065">K2560065 </a> </td>
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                        <td>5'Con1</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560066">K2560066 </a></td>
 +
                        <td>5'Con2</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
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<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560067">K2560067 </a></td>
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                        <td>5'Con3</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560066">K2560066 </a></td>
 +
                        <td>5'Con4</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560069">K2560069 </a></td>
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                        <td>5'Con5</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
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<tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560075">K2560075 </a></td>
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                        <td>5'Con1 Short Res </td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
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<tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560076">K2560076 </a></td>
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                        <td>5'Con2 Short</td>
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                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
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                    </tr>
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<tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560077">K2560077 </a></td>
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                        <td>5'Con3 Short</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560078">K2560078 </a></td>
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                        <td>5'Con4 Short</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560079">K2560079 </a></td>
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                        <td>5'Con5 Short</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560095">K2560095 </a></td>
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                        <td>5'Con1 inv</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560098">K2560096 </a></td>
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                        <td>5'Con2 inv</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560097">K2560097 </a></td>
 +
                        <td>5'Con3 inv</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560098">K2560098 </a></td>
 +
                        <td>5'Con4 inv</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560099">K2560099 </a></td>
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                        <td>5'Con5 inv</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560105">K2560105 </a></td>
 +
                        <td>5'Con5 inv Ori</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560107">K2560107 </a></td>
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                        <td>5'Con1 Res</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
                    </table>
 +
 +
 +
</div>
 +
</div>
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 +
 +
 +
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 +
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 +
<div class="collapsible">
 +
<div class="btn_expand">Promoter</div>
 +
<div class="content">
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 +
<div>
 +
 +
 +
 +
<div>  <a href=https://static.igem.org/mediawiki/parts/f/f4/T--Marburg--Promoters.zip>Download all Promoters</abbr></a>
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 +
</div>
 +
<div>
 +
 +
<form action="https://static.igem.org/mediawiki/parts/f/f4/T--Marburg--Promoters.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
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 +
</form>
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 +
</div>
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<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
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                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                     
 +
                    </tr>
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                </thead>
 +
                <tbody>
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 +
 +
<tr>
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                        <td><a href="http://parts.igem.org/Part:BBa_K2560007">K2560007 </a></td>
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                        <td>J23100</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
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<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560009">K2560009 </a></td>
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                        <td>J23104</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560014">K2560014 </a></td>
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                        <td>J23106</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560015">K2560015 </a></td>
 +
                        <td>J23115</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560017">K2560017 </a></td>
 +
                        <td>J23101</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560018">K2560018 </a></td>
 +
                        <td>J23102</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560019">K2560019 </a></td>
 +
                        <td>J23103</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560020">K2560020 </a></td>
 +
                        <td>J23105</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560021">K2560021 </a></td>
 +
                        <td>J23107</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560022">K2560022 </a></td>
 +
                        <td>J23108</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560023">K2560023 </a></td>
 +
                        <td>J23109</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560024">K2560024 </a></td>
 +
                        <td>J23110</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560025">K2560025 </a></td>
 +
                        <td>J23111</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560026">K2560026 </a></td>
 +
                        <td>J23113</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560027">K2560027 </a></td>
 +
                        <td>J23114</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560028">K2560028 </a></td>
 +
                        <td>J23116</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560029">K2560029 </a></td>
 +
                        <td>J23117</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560029">K2560029 </a></td>
 +
                        <td>J23117</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560030">K2560030 </a></td>
 +
                        <td>J23118</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560031">K2560031 </a></td>
 +
                        <td>J23119</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560123">K2560123 </a></td>
 +
                        <td>pTet</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560124">K2560124 </a></td>
 +
                        <td>pTrc</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560131">K2560131 </a></td>
 +
                        <td>Promoter Dummy</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
                    </table>
 +
 +
 +
</div>
 +
</div>
 +
 +
 +
 +
<div class="collapsible">
 +
<div class="btn_expand">RBS</div>
 +
<div class="content">
 +
 +
 +
 +
 +
<div>  <a href=https://static.igem.org/mediawiki/parts/3/33/T--Marburg--RBS%27s.zip>Download all RBS's</abbr></a>
 +
 +
</div>
 +
<div>
 +
 +
<form action="https://static.igem.org/mediawiki/parts/3/33/T--Marburg--RBS%27s.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
 +
 +
</form>
 +
 +
</div>
 +
 +
 +
 +
 +
<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
 +
                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                       
 +
                    </tr>
 +
                </thead>
 +
                <tbody>
 +
 +
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560008">K2560008 </a></td>
 +
                        <td>B0034</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560010">K2560010 </a></td>
 +
                        <td>B0032</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560013">K2560013 </a></td>
 +
                        <td>B0031</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560016">K2560016 </a></td>
 +
                        <td>B0030</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560084">K2560084 </a></td>
 +
                        <td>RBS Dummy</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
                    </table>
 +
 +
</div>
 +
</div>
 +
 +
 +
 +
 +
 +
 +
 +
<div class="collapsible">
 +
<div class="btn_expand">CDS</div>
 +
<div class="content">
 +
 +
 +
 +
 +
<div>  <a href=https://static.igem.org/mediawiki/parts/d/d9/T--Marburg--CDS%27s.zip>Download all CDS's</abbr></a>
 +
 +
</div>
 +
<div>
 +
 +
<form action="https://static.igem.org/mediawiki/parts/d/d9/T--Marburg--CDS%27s.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
 +
 +
</form>
 +
 +
</div>
 +
 +
 +
 +
<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
 +
                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                     
 +
                    </tr>
 +
                </thead>
 +
                <tbody>
 +
 +
 +
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560033">K2560033 </a></td>
 +
                        <td>E1010</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560041">K2560041 </a></td>
 +
                        <td>E0030</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560042">K2560042 </a></td>
 +
                        <td>sfGFP</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560043">K2560043 </a></td>
 +
                        <td>sfGFP Vn</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560047">K2560047 </a></td>
 +
                        <td>Cas9</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560051">K2560051 </a></td>
 +
                        <td>Lux Operon</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560054">K2560054 </a></td>
 +
                        <td>dCas9</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560082">K2560082 </a></td>
 +
                        <td>CDS Dummy</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560114">K2560114 </a></td>
 +
                        <td>LacZ Alpha</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560128">K2560128 </a></td>
 +
                        <td>K6600004</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560135">K2560135 </a></td>
 +
                        <td>SXT Beta</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560268">K2560268 </a></td>
 +
                        <td>tfox Vc</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560269">K2560269</a></td>
 +
                        <td>tfox Vn</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560270">K2560270 </a></td>
 +
                        <td>SXT</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560272">K2560272 </a></td>
 +
                        <td>flp recombinase</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
                    </table>
 +
 +
 +
</div>
 +
</div>
 +
 +
 +
 +
 +
 +
 +
 +
 +
<div class="collapsible">
 +
<div class="btn_expand">Terminator</div>
 +
<div class="content">
 +
 +
 +
 +
<div>  <a href=https://static.igem.org/mediawiki/parts/1/18/T--Marburg--Terminators.zip>Download all Terminators</abbr></a>
 +
 +
</div>
 +
<div>
 +
 +
<form action="https://static.igem.org/mediawiki/parts/1/18/T--Marburg--Terminators.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
 +
 +
</form>
 +
 +
</div>
 +
 +
 +
 +
 +
 +
 +
 +
<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
 +
                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                       
 +
                    </tr>
 +
                </thead>
 +
                <tbody>
 +
 +
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560034">K2560034 </a></td>
 +
                        <td>B0010</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560035">K2560035 </a></td>
 +
                        <td>B0015</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560081">K2560081 </a></td>
 +
                        <td>Terminator Dummy</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560089">K2560089 </a></td>
 +
                        <td>B1002</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560091">K2560091 </a></td>
 +
                        <td>B0003</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560093">K2560093 </a></td>
 +
                        <td>B0006</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
                    </table>
 +
 +
</div>
 +
</div>
 +
 +
 +
 +
<div>
 +
 +
 +
 +
<div class="collapsible">
 +
<div class="btn_expand">3'Connector</div>
 +
<div class="content">
 +
 +
 +
 +
 +
<div>  <a href=https://static.igem.org/mediawiki/parts/3/3c/T--Marburg--3%27Connectors.zip>Download all 3'Connectors</abbr></a>
 +
 +
</div>
 +
<div>
 +
 +
<form action="https://static.igem.org/mediawiki/parts/3/3c/T--Marburg--3%27Connectors.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
 +
 +
</form>
 +
 +
</div>
 +
 +
 +
 +
 +
<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
 +
                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                     
 +
                    </tr>
 +
                </thead>
 +
                <tbody>
 +
 +
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560012">K2560012 </a></td>
 +
                        <td>3'Connector Dummy</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560070">K2560070 </a></td>
 +
                        <td>3'Con1</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560071">K2560071 </a></td>
 +
                        <td>3'Con2</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560072">K2560072 </a></td>
 +
                        <td>3'Con3</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560073">K2560073 </a></td>
 +
                        <td>3'Con4</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560080">K2560080 </a></td>
 +
                        <td>3'Con5 Ori</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560100">K2560100 </a></td>
 +
                        <td>3'Con1 inv Short</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560101">K2560101 </a></td>
 +
                        <td>3'Con2 inv Short</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560102">K2560102 </a></td>
 +
                        <td>3'Con3 inv Short</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560103">K2560103 </a></td>
 +
                        <td>3'Con4 inv Short</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560104">K2560104 </a></td>
 +
                        <td>3'Con5 inv Short</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560106">K2560106 </a></td>
 +
                        <td>3'Con1 inv Res</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560108">K2560108 </a></td>
 +
                        <td>3'Con1 inv</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560109">K2560109 </a></td>
 +
                        <td>3'Con1 inv Res</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560110">K2560110 </a></td>
 +
                        <td>3'Con2 inv</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560111">K2560111 </a></td>
 +
                        <td>3'Con3 inv </td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560112">K2560112 </a></td>
 +
                        <td>3'Con4 inv </td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560113">K2560113 </a></td>
 +
                        <td>3'Con5 inv </td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>           
 +
                    </table>
 +
 +
</div>
 +
</div>
 +
 +
 +
 +
 +
 +
 +
 +
 +
 +
 +
<div class="collapsible">
 +
<div class="btn_expand">ORI</div>
 +
<div class="content">
 +
 +
 +
<div>  <a href=https://static.igem.org/mediawiki/parts/2/21/T--Marburg--ORIs.zip>Download all ORIs</abbr></a>
 +
 +
</div>
 +
<div>
 +
 +
<form action="https://static.igem.org/mediawiki/parts/2/21/T--Marburg--ORIs.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
 +
 +
</form>
 +
 +
</div>
 +
 +
 +
 +
 +
<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
 +
                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                   
 +
                    </tr>
 +
                </thead>
 +
                <tbody>
 +
 +
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560036">K2560036 </a></td>
 +
                        <td>ColE1</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560037">K2560037 </a></td>
 +
                        <td>pMB1</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560046">K2560046 </a></td>
 +
                        <td>p15A</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
                    </table>
 +
   
 +
</div>
 +
</div>
 +
 +
 +
 +
 +
<div class="collapsible">
 +
<div class="btn_expand">Antibiotic Resistance</div>
 +
<div class="content">
 +
 +
 +
 +
<div>  <a href=https://static.igem.org/mediawiki/parts/b/bd/T--Marburg--Not_Antibiotic_Resistance.zip>Download all Antibiotic Resistances</abbr></a>
 +
 +
</div>
 +
<div>
 +
 +
<form action="https://static.igem.org/mediawiki/parts/b/bd/T--Marburg--Not_Antibiotic_Resistance.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
 +
 +
</form>
 +
 +
</div>
 +
 +
 +
 +
 +
 +
<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
 +
                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                     
 +
                    </tr>
 +
                </thead>
 +
                <tbody>
 +
 +
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560048">K2560048 </a></td>
 +
                        <td>Cam. Res. RFP</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560056">K2560056 </a></td>
 +
                        <td>Kan. Res.(pSB3K3) RFP</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560057">K2560057 </a></td>
 +
                        <td>Kan. Res. (pSB3K3) GFP</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560058">K2560058 </a></td>
 +
                        <td>Tet. Res. (pSB3T5) RFP</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560059">K2560059 </a></td>
 +
                        <td>Tet. Res. (pSB3T5) GFP</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560125">K2560125 </a></td>
 +
                        <td>Carb. Res. RFP</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560126">K2560126 </a></td>
 +
                        <td>Carb. Res. GFP</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560127">K2560127 </a></td>
 +
                        <td>Carb. Res. into BBa_K2560002</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560132">K2560132 </a></td>
 +
                        <td>Cam. Res. into BBa_K2560002</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560133">K2560133 </a></td>
 +
                        <td>Kan. Res. into BBa_K2560002</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560134">K2560134 </a></td>
 +
                        <td>Tet. Res. into BBa_K2560002</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
                    </table>
 +
 +
</div>
 +
</div>
 +
 +
 +
 +
 +
 +
 +
<div class="collapsible">
 +
<div class="btn_expand">N-Tag</div>
 +
<div class="content">
 +
 +
 +
 +
<div>  <a href=https://static.igem.org/mediawiki/parts/d/d8/T--Marburg--N-Tags.zip>Download all N-Tags</abbr></a>
 +
 +
</div>
 +
<div>
 +
 +
<form action="https://static.igem.org/mediawiki/parts/d/d8/T--Marburg--N-Tags.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
 +
 +
</form>
 +
 +
</div>
 +
 +
 +
 +
 +
<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
 +
                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                     
 +
                    </tr>
 +
                </thead>
 +
                <tbody>
 +
 +
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560060">K2560060 </a></td>
 +
                        <td>E1010 4x</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560063">K2560063 </a></td>
 +
                        <td>sfGFP 4x</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560085">K2560085 </a></td>
 +
                        <td>His Tag 4x</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560087">K2560087 </a></td>
 +
                        <td>FLAG Tag 4x</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560129">K2560129 </a></td>
 +
                        <td>K660004 4x</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560257">K2560257 </a></td>
 +
                        <td>Streptag 4x</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
 +
      </table>
 +
 +
</div>
 +
</div>
 +
 +
 +
 +
 +
 +
 +
 +
<div class="collapsible">
 +
<div class="btn_expand">C-Tag</div>
 +
<div class="content">
 +
 +
<div>  <a href=https://static.igem.org/mediawiki/parts/4/44/T--Marburg--C-Tags.zip>Download all C-Tags</abbr></a>
 +
 +
</div>
 +
<div>
 +
 +
<form action="https://static.igem.org/mediawiki/parts/4/44/T--Marburg--C-Tags.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
 +
 +
</form>
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</div>
 +
 +
 +
 +
 +
<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
 +
                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                       
 +
                    </tr>
 +
                </thead>
 +
                <tbody>
 +
 +
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560061">K2560061 </a></td>
 +
                        <td>E1010 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560062">K2560062 </a></td>
 +
                        <td>B0015 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560064">K2560064 </a></td>
 +
                        <td>sfGFP 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560086">K2560086 </a></td>
 +
                        <td>His Tag 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560088">K2560088 </a></td>
 +
                        <td>FLAG Tag 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560090">K2560090 </a></td>
 +
                        <td>B1002 5b</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560092">K2560092 </a></td>
 +
                        <td>B1003 5b</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560094">K2560094 </a></td>
 +
                        <td>B1006 5b</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560117">K2560117 </a></td>
 +
                        <td>I11012 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560118">K2560118 </a></td>
 +
                        <td>M0050 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560119">K2560119 </a></td>
 +
                        <td>M0051 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560130">K2560130 </a></td>
 +
                        <td>K660004 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560258">K2560258 </a></td>
 +
                        <td>Streptag 5a</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
 +
      </table>
 +
 +
</div>
 +
</div>
 +
 +
 +
 +
 +
 +
 +
<div class="collapsible">
 +
<div class="btn_expand">Entry Vector</div>
 +
<div class="content">
 +
 +
<div>  <a href=http://https://static.igem.org/mediawiki/parts/4/46/T--Marburg--Entry_Vectors.zip>Download all Entry Vectors</abbr></a>
 +
 +
</div>
 +
<div>
 +
 +
<form action="http://https://static.igem.org/mediawiki/parts/4/46/T--Marburg--Entry_Vectors.zip" method="get" target="_blank">
 +
<input type="image" src="https://static.igem.org/mediawiki/parts/c/c8/T--Marburg--Download_icon.svg" width=100 height=100>
 +
 +
</form>
 +
 +
</div>
 +
 +
 +
 +
<table>
 +
                <thead>
 +
                    <tr>
 +
                        <th> BioBrick</th>
 +
                        <th> Name</th>
 +
                        <th> Plasmid</th>
 +
                       
 +
                    </tr>
 +
                </thead>
 +
                <tbody>
 +
 +
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560001">K2560001 </a></td>
 +
                        <td>Entry Vector with RFP Dropout</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560002">K2560002 </a></td>
 +
                        <td>Entry Vector with GFP Dropout</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560005">K2560005 </a></td>
 +
                        <td>Resistance Entry Vector with RFP Dropout</td>
 +
                        <td>Falsch</td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560006">K2560006 </a></td>
 +
                        <td>Resistance Entry Vector with GFP Dropout</td>
 +
                        <td>Falsch</td>
 +
                    </tr>
 +
<tr>
 +
                        <td><a href="http://parts.igem.org/Part:BBa_K2560305">K2560305 </a></td>
 +
                        <td>gRNA Entry Vector with GFP Dropout</td>
 +
                        <td><a href=http://parts.igem.org/Part:BBa_K2560002>BBa_K2560002 (pSB1C3 derivative)</a></td>
 +
                    </tr>
 +
 +
      </table>
 +
 +
</div>
 
</div>
 
</div>
  

Latest revision as of 02:49, 18 October 2018

Parts Overview
Best Basic Part
As best basic part we woud like to present tfoX, the inducer of natural competence in Vibrio species.
Since scientific progress keeps accelerating in the past years, the fast production of results gains more and more importance. We are convinced that in the near future the community of synthetic biology will replace E. coli by the much faster growing organism V. natriegens. We strongly believe that our best basic part tfoX will support you in speeding up your research. Making this part accessible for everyone we would like to accelerate engineering biology and help you to make the world a better place. Tfox is known as the inducer of natural competence in some Vibrio species like V. natriegens. This ability of cells to take up free DNA is called competence. A special kind is the natural competence. In 1928, natural competence was first observed by Frederick Griffith even if he did not understand the genetics behind (Griffithet al.1928) . Thanks to many more great scientists who did research in this area, we now know a lot about natural competence and the ability of some bacteria to take up free DNA.
In their natural environment, the regulator protein Tfox activates Type VI secretion systems (T6SSs) and allows bacteria to penetrate prey cells, leading to cell lysis and the release of DNA. The naturally competent cells can take up the free DNA from their environment and utilize it for genomic modifications by homologous recombination into their genome. Although much progresses has been made in this area, the molecular details of natural competence are not understood completely (Wuet al.2015) . For V. cholerae, it is known, that the expression of tfoX is induced by surface contact to chitin (Figure 1). Even though the V. natriegens genome is encoding a tfoX gene, its natural inducer is not known until now. However it is known, that the induction of expression of the V. cholerae tfoX is leading to competence in V. natriegens cells as well. (Hayes et al. 2017).
Schematic figure on mode of action of TfoX.
Figure 1: Schematic figure on mode of action of TfoX in V. cholerae. When cells get in contact with chitin, tfoX is expressed. Since TfoX is a positive regulator of natural competence, DNA from the environment is taken up into the cell.
We compared the domains of TfoX with the domains of homologous proteins. These domains are listed below and in Figure 2.

  • TfoX, N-terminal (TfoX_N): N-terminal domain of TfoX found in Haemophilus influenza, also present in Sxy found in E. coli (inducer of natural competence)
  • TfoX, C-terminal (TfoX_C): C-terminal domain of TfoX found in Haemophilus influenza. This family corresponds to the C-terminal presumed domain of TfoX. The domain is found in association with the N-terminal domain in some, but not all members of this group, suggesting this is an autonomous and functionally unrelated domain.
  • Coiled Coil region (CC): Coiled Coil domain. This family is found in many proteins. Sometimes they are responsible for self-interaction, sometimes they are used to keep distance to other proteins.
Schematic figure on mode of action of TfoX.
Figure 1: Schematic figure on mode of action of TfoX in V. cholerae. When cells get in contact with chitin, tfoX is expressed. Since TfoX is a positive regulator of natural competence, DNA from the environment is taken up into the cell.
Since we are convinced, that E. coli will be replaced by V. natriegens in the next few years, we believe that the usage of natural transformation will become more significant. There are several advantages of this method in comparison to electroporation or heat shock mediated transformation. First, it is less stressful to the cells and second, you do not need to aliquot a thousands of chemically competent E. colis in Eppendorf cups every few weeks. If you are planning to do a transformation, the only thing to do is to inoculate the desired V. natriegens strain in a liquid culture, inducing the expression of tfoX (Figure 3). Additionally, modified V. natriegens strains can be produced much faster, due to the possibility to transform linear fragments for genomic modifications instead of wasting time with cloning plasmids. Even more linear fragments can be used for co-transformation, creating several genomic modifications at the same time (Hayes et al. 2017) .
In the future natural transformation could be utilized in combination with genome engineering methods like MAGE and CAGE, which rely on electroporation for E.coli, that could allow more rapid genome designs, such as complete codon reassignments.

Schematic figure on mode of action of TfoX.
Figure 3:Workflow of a natural transformation.

Click here to download the protocol we used for natural transformation

Providing this basic part, we want to encourage the community of iGEM as well as all other scientists in the field of synthetic biology to start working with V. natriegens. Including tfoX into the modular cloning system of iGEM and the Marburg Collection we hope to usher in a new era of synthetic biology. We would love to see future iGEM teams using this basic part for further constructing their individual transcription unit variants in order to screen for the maximum efficiency for customized applications.

Using tfox in full transcriptional units, we were able to perform natural transformation with plasmids (Figure 4) as well as with linear DNA to create our strains with genomic modifications.
Plates showing successful transformation of pYTK.
Figure 4: Plates showing the successful transformation of plasmids using natural transformation. Left plate is showingtransformation of cells with pYTK. Right plate is showing negative control.
Click here to learn more about how we used our best basic part in our project on the following sites in the Strain Engineering sub-project:
Best Composite Part
As best composite part we extended our best basic part TfoX, the inducer of natural competence in Vibrio species. We could successfully combine it with one of the inducible promoters from our Marburg Collection.
Relative promoter strength of Anderson promoters
Figure 1:Alignment of sequencing results of our first constructed Lvl1 tfoX plasmids showing missing basepairs at places at which black fragments are missing.

As an extension of our basic part tfoX we created a full transcriptional unit enabling you to perform natural transformation in V. natriegens. First we started assembling a full transcriptional unit using a constitutive promoter. Therefore, we assembled tfoX together with the constitutive promoters J23100 or J23110 into a transcriptional unit by golden gate assembly. After transformation of this reactions into E. coli we were able to get colonies that seemed to be positive for the Lvl1 plasmids encoding tfoX because they showed the right size of fragment after plasmid preparation and test digestion, whenever we sent them for sequencing, some basepairs of the tfox gene were always missing (Figure 1).

Alignment of sequencing results of our first constructed Lvl1 <i>tfoX</i> plasmids showing missing basepairs.
Figure 2: Relative promoter strength of Anderson promoters Data were normalized over the strongest construct J23100. Error bars represent the standard deviation of the measurements of three independent experiments
We suggest that these problems could be caused because Tfox could somehow ne toxic to E. coli cells. At this point of our project we realized how important it is to have well characterized, tight, inducible promoters for the usage in V. natriegens. In addition to constitutive promoters, the Marburg Collection contains the inducible promoters, pTet and pTrc. From our measurements of the promoter characterization, we learned that the pTet promoter is the tightest one showing the lowest value of Luminescence without induction (Figure 2). The pTet promoter can be induced by the tetracycline derivative anhydrotetracycline (ATc). ATc is much less cytotoxic than IPTG but still capable of binding and altering the structure of the repressor TetR, leading to release of the promoter and enabling transcription.
1 %- Agarose gel of the test digest.
Figure 3:1 %- Agarose gel of the test digest. 2-logDNA ladder is used as marker on agarose gels.

Using this suitable promoter for V. natriegens we successfully assembled a plasmid containing a full transcriptional unit with a pTet promoter and did not had any problems during the cloning process. By Test Digestion and sequencing we could prove, that now the sequence of Tfox was correct (Figure 3 and 4).

Alignment of sequencing results of our final constructed Lvl1 <i>tfoX</i> plasmid showing the correct sequence.
Figure 4:Alignment of sequencing results of our final constructed Lvl1 tfoX plasmid showing the correct sequence.
As we have already shown in the chapter of our basic part, we were able to perform natural transformation using this plasmid and could demonstrate, that GFP which is encoded on pYTK is expressed by V. natriegens.
Plates showing successful transformation of pYTK.
Figure 5:Plate showing a successful transformation of pYTK using natural transformation.
Plates showing successful transformation of pYTK.
Figure 6:Plate showing a successful transformation of pYTK using natural transformation with functioning pYTK.
Click here to learn more about how we used our best composite part in our project on the following sites in the Strain Engineering sub-project:
Marburg Collection
We created a novel golden-gate based cloning toolbox consisting of 123 parts. Plasmid maps of all parts can be downloaded here.

Parts of the Marburg Collection

Promoter
Entry Vector
BioBrick Name Plasmid
K2560001 Entry Vector with RFP Dropout BBa_K2560002 (pSB1C3 derivative)
K2560002 Entry Vector with GFP Dropout BBa_K2560002 (pSB1C3 derivative)
K2560005 Resistance Entry Vector with RFP Dropout Falsch
K2560006 Resistance Entry Vector with GFP Dropout Falsch
K2560305 gRNA Entry Vector with GFP Dropout BBa_K2560002 (pSB1C3 derivative)
B. Marchal