Difference between revisions of "Team:TECHNION-ISRAEL/Basic Part"

 
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<h1> Basic Parts </h1>
 
<h1> Basic Parts </h1>
 
 
<img src="https://static.igem.org/mediawiki/2017/f/f9/T--TECHNION-ISRAEL--co-assay.png" class="cover" alt=""  style= "width:18% ; margin: auto;">
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<img src="https://static.igem.org/mediawiki/2017/8/81/T--TECHNION-ISRAEL--basic_part.png" class="cover" alt=""  style= "width:15% ; margin: auto;">
 
 
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<strong> Mutated EF-1a promoter </strong> is a strong, constitutive, promoter in mammalian cells. It is particularly useful in cells that do not express proteins with other common promoters (such as CMV). This promoter is commonly used in stem cells as it is one of the few promoters that prompt high levels of protein expression in these predominantly dormant cells.
 
 
<br>
 
<br>
This promoter is our favorite part.  Promoters are clearly very important in iGEM. Synthetic biology depends on strong and consistent expression of recombinant proteins.  The EF-1a promoter was used by both teams who won “best therapeutics project” in 2016 (LINK TO TEL HAI, LINK TO SYSU MED 2016), but neither submitted it as a part since the promoter contains two forbidden restriction sites within its sequence. As a side project, we set out to create a mutated version of the original (WT) EF-1a so that it may serve future iGEM teams. We mutated the WT EF-1a at two points (base pair 319 and 824 were changed from C to T), thus eliminating the forbidden restriction sites. We characterized and tested our new promoter for functionality and compared it with the WT EF-1a (Figure 1). In closing, we have added a new and valuable promoter to the iGEM parts registry that we believe will serve future teams seeking to work with mammalian cells.  
+
<strong> Mutated EF-1a promoter </strong> (<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520023">BBa_K2520023</a>) is a strong, constitutive, promoter in mammalian cells. It is particularly useful in cells that do not express proteins with other common promoters (such as CMV). This promoter is commonly used in stem cells as it is one of the few promoters that prompt high levels of protein expression in these predominantly dormant cells.
 +
<br>
 +
This promoter is our favorite part.  Promoters are clearly very important in iGEM. Synthetic biology depends on strong and consistent expression of recombinant proteins.  The EF-1a promoter was used by both teams who won “best therapeutics project” in 2016 (<a target="_blank" href="https://2016.igem.org/Team:Tel-Hai">Tel-Hai</a>, <a target="_blank" href="https://2016.igem.org/Team:SYSU-MEDICINE">SYSU MEDICINE</a>), but neither submitted it as a part since the promoter contains two forbidden restriction sites within its sequence. As a side project, we set out to create a mutated version of the original (WT) EF-1a so that it may serve future iGEM teams. We mutated the WT EF-1a at two points (base pair 319 and 824 were changed from C to T), thus eliminating the forbidden restriction sites. We characterized and tested our new promoter for functionality and compared it with the WT EF-1a (Figure 1 at the <a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520023">part's page</a>). In closing, we have added a new and valuable promoter to the iGEM parts registry that we believe will serve future teams seeking to work with mammalian cells.  
 
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<br>
 
<br>
 
<br>
  
<strong> Secrecon </strong> is an artificial secretory signal, which leads to high levels of protein secretion and membrane expression. Secrecon was found to be the most powerful and consistent human secretion signal sequence (more than Igk, for example).  
+
<strong> Secrecon </strong> (<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520002">BBa_K2520002</a>) is an artificial secretory signal, which leads to high levels of protein secretion and membrane expression. Secrecon was found to be the most powerful and consistent human secretion signal sequence (more than Igk, for example).  
  
 
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<br>
<strong> B7-1 </strong> is a trans-membrane domain protein with a cytoplasmic tail. It enhances transport from the endoplasmic reticulum and allows accumulation of chimeric proteins on the plasma membrane. It has been shown to allow for the most consistent and long lasting membrane expression in human cells (more than PDGFR, for example).  
+
<strong> B7-1 </strong> (<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520000">BBa_K2520000</a>) is a trans-membrane domain protein with a cytoplasmic tail. It enhances transport from the endoplasmic reticulum and allows accumulation of chimeric proteins on the plasma membrane. It has been shown to allow for the most consistent and long lasting membrane expression in human cells (more than PDGFR, for example).  
  
 
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<strong> Rat anti-mouse IgM scFv </strong> is a synthetic version of an anti-mouse IgM antibody. It binds the constant heavy region of all mouse antibodies with the IgM isotype. This recombinant protein can be used to bind primary (IgM) antibodies from murine source.
+
<strong> Rat anti-mouse IgM scFv </strong> (<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520001">BBa_K2520001</a>) is a synthetic version of an anti-mouse IgM antibody. It binds the constant heavy region of all mouse antibodies with the IgM isotype. This recombinant protein can be used to bind primary (IgM) antibodies from murine source.
  
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520000" target="_blank">BBa_K2520000</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520000" target="_blank">BBa_K2520000</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520001" target="_blank">BBa_K2520001</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520001" target="_blank">BBa_K2520001</a>
 
</td>
 
</td>
 
                             <td>Coding</td>
 
                             <td>Coding</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520002" target="_blank">BBa_K2520002
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520002" target="_blank">BBa_K2520002
  
 
</a>
 
</a>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520023" target="_blank">BBa_K2520023</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520023" target="_blank">BBa_K2520023</a>
 
</td>
 
</td>
 
                             <td>Regulatory</td>
 
                             <td>Regulatory</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520024" target="_blank">BBa_K2520024</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520024" target="_blank">BBa_K2520024</a>
 
</td>
 
</td>
 
                             <td>Signalling</td>
 
                             <td>Signalling</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520025" target="_blank">BBa_K2520025</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520025" target="_blank">BBa_K2520025</a>
 
</td>
 
</td>
 
                             <td>Regulatory</td>
 
                             <td>Regulatory</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520026" target="_blank">BBa_K2520026</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520026" target="_blank">BBa_K2520026</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520027" target="_blank">BBa_K2520027</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520027" target="_blank">BBa_K2520027</a>
 
</td>
 
</td>
 
                             <td>Coding</td>
 
                             <td>Coding</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520028" target="_blank">BBa_K2520028</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520028" target="_blank">BBa_K2520028</a>
 
</td>
 
</td>
 
                             <td>Signalling</td>
 
                             <td>Signalling</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520029" target="_blank">BBa_K2520029</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520029" target="_blank">BBa_K2520029</a>
 
</td>
 
</td>
 
                             <td>Signalling</td>
 
                             <td>Signalling</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520030" target="_blank">BBa_K2520030</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520030" target="_blank">BBa_K2520030</a>
 
</td>
 
</td>
 
                             <td>Tag</td>
 
                             <td>Tag</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520031" target="_blank">BBa_K2520031</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520031" target="_blank">BBa_K2520031</a>
 
</td>
 
</td>
 
                             <td>Tag</td>
 
                             <td>Tag</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520032" target="_blank">BBa_K2520032</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520032" target="_blank">BBa_K2520032</a>
 
</td>
 
</td>
 
                             <td>Tag</td>
 
                             <td>Tag</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520033" target="_blank">BBa_K2520033</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520033" target="_blank">BBa_K2520033</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td>Noa Eden</td>
 
                             <td>Noa Eden</td>
 
<td>66</td>
 
<td>66</td>
                        </tr>
 
 
                        <tr>
 
                            <td></td>
 
                            <td></td>
 
                            <td></td>
 
                            <td>
 
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520034" target="_blank">BBa_K2520034</a>
 
</td>
 
                            <td>Protein_Domain</td>
 
                            <td>T2A</td>
 
                            <td>Noa Eden</td>
 
<td>63</td>
 
 
                         </tr>
 
                         </tr>
 
 
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520036" target="_blank">BBa_K2520036</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520036" target="_blank">BBa_K2520036</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520037" target="_blank">BBa_K2520037</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520037" target="_blank">BBa_K2520037</a>
 
</td>
 
</td>
 
                           <td>Protein_Domain</td>
 
                           <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520038" target="_blank">BBa_K2520038</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520038" target="_blank">BBa_K2520038</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520039" target="_blank">BBa_K2520039</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520039" target="_blank">BBa_K2520039</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520040" target="_blank">BBa_K2520040</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520040" target="_blank">BBa_K2520040</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520041" target="_blank">BBa_K2520041</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520041" target="_blank">BBa_K2520041</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520042" target="_blank">BBa_K2520042</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520042" target="_blank">BBa_K2520042</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520043" target="_blank">BBa_K2520043</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520043" target="_blank">BBa_K2520043</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520044" target="_blank">BBa_K2520044</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520044" target="_blank">BBa_K2520044</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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                             <td></td>
 
                             <td></td>
 
                             <td>
 
                             <td>
<a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520045" target="_blank">BBa_K2520045</a>
+
<a target="_blank" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2520045" target="_blank">BBa_K2520045</a>
 
</td>
 
</td>
 
                             <td>Protein_Domain</td>
 
                             <td>Protein_Domain</td>
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Latest revision as of 02:21, 2 November 2017

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Basic Parts

Basic Parts




We designed four new basic parts that were used in our project:

Mutated EF-1a promoter (BBa_K2520023) is a strong, constitutive, promoter in mammalian cells. It is particularly useful in cells that do not express proteins with other common promoters (such as CMV). This promoter is commonly used in stem cells as it is one of the few promoters that prompt high levels of protein expression in these predominantly dormant cells.
This promoter is our favorite part. Promoters are clearly very important in iGEM. Synthetic biology depends on strong and consistent expression of recombinant proteins. The EF-1a promoter was used by both teams who won “best therapeutics project” in 2016 (Tel-Hai, SYSU MEDICINE), but neither submitted it as a part since the promoter contains two forbidden restriction sites within its sequence. As a side project, we set out to create a mutated version of the original (WT) EF-1a so that it may serve future iGEM teams. We mutated the WT EF-1a at two points (base pair 319 and 824 were changed from C to T), thus eliminating the forbidden restriction sites. We characterized and tested our new promoter for functionality and compared it with the WT EF-1a (Figure 1 at the part's page). In closing, we have added a new and valuable promoter to the iGEM parts registry that we believe will serve future teams seeking to work with mammalian cells.

Secrecon (BBa_K2520002) is an artificial secretory signal, which leads to high levels of protein secretion and membrane expression. Secrecon was found to be the most powerful and consistent human secretion signal sequence (more than Igk, for example).

B7-1 (BBa_K2520000) is a trans-membrane domain protein with a cytoplasmic tail. It enhances transport from the endoplasmic reticulum and allows accumulation of chimeric proteins on the plasma membrane. It has been shown to allow for the most consistent and long lasting membrane expression in human cells (more than PDGFR, for example).

Rat anti-mouse IgM scFv (BBa_K2520001) is a synthetic version of an anti-mouse IgM antibody. It binds the constant heavy region of all mouse antibodies with the IgM isotype. This recombinant protein can be used to bind primary (IgM) antibodies from murine source.

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