Difference between revisions of "Team:UCLouvain/Basic Part"
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<div class="page-title"> | <div class="page-title"> | ||
| − | <div class="big-title montserrat-text uppercase"> | + | <div class="big-title montserrat-text uppercase">Parts</div> |
| − | <div class="small-title montserrat-text uppercase"><br><Marine style="margin-left:32px;font-family:'Brush Script MT';font-size:32px;text-transform:none;font-weight:bold;"> | + | <div class="small-title montserrat-text uppercase"><br><Marine style="margin-left:32px;font-family:'Brush Script MT';font-size:32px;text-transform:none;font-weight:bold;">BactaSun</Marine></div> |
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<div class="row" style="margin-bottom: 40px;"> | <div class="row" style="margin-bottom: 40px;"> | ||
| − | <div class="col-md- | + | <div class="col-md-6"> |
| − | + | <div class="section-title" style="text-align:center;float:left;width:100%;margin-bottom:0"> | |
| − | <span> | + | <span>Auxotrophic Approach</span> |
| − | </div> | + | </div> |
| + | <BR><BR> | ||
| + | <div class="section-title" style="text-align:center;float:left;width:100%;margin-bottom:0"> | ||
| + | <span><Marine style="margin-left:32px;font-family:'Brush Script MT';font-size:32px;text-transform:none;font-weight:bold;">Basic parts</Marine></span> | ||
| + | </div> | ||
| + | <BR><BR> | ||
| + | |||
| + | <div class="section-title" style="text-align:center;float:left;width:100%;margin-bottom:0"> | ||
| + | <p><i><center>Source : iGEM DNA Distribution Kit</center></i></p> | ||
| + | </div> | ||
| + | <BR><BR> | ||
| + | |||
| + | <div class="section-title" style="text-align:left;float:left;width:100%;margin-bottom:0"> | ||
| + | <p class="montserrat-text uppercase">pBAD/araC <a style="color:#0066cc;" href="http://parts.igem.org/Part:BBa_I0500">(I0500)</a></p> | ||
| + | |||
| + | <p>The pBAD promoter is a very common inducible one that is activated by L-arabinose and repressed by araC.</p> | ||
| + | |||
<BR><BR> | <BR><BR> | ||
| − | + | <div class="section-title" style="text-align:left;float:left;width:100%;margin-bottom:0"> | |
| − | <p class="montserrat-text uppercase"> | + | <p class="montserrat-text uppercase">Terminator <a style="color:#0066cc;" href="http://parts.igem.org/Part:BBa_B0010">(B0010)</a></p> |
| − | + | </div> | |
| − | + | <p>Terminators are necessary to stop the transcription at the end of a coding sequence.</p> | |
<BR><BR> | <BR><BR> | ||
| − | + | <div class="section-title" style="text-align:left;float:left;width:100%;margin-bottom:0"> | |
| − | <p class="montserrat-text uppercase"> | + | <p class="montserrat-text uppercase">RBS <a style="color:#0066cc;" href="http://parts.igem.org/Part:BBa_B0034">(B0034)</a></p> |
| − | + | </div> | |
| − | + | <p>Efficient ribosome binding site characterized by IIT Madras in iGEM 2016.</p> | |
<BR><BR> | <BR><BR> | ||
| − | + | <div class="section-title" style="text-align:left;float:left;width:100%;margin-bottom:0"> | |
| − | <p class="montserrat-text uppercase"> | + | <p class="montserrat-text uppercase">RFP <a style="color:#0066cc;" href="http://parts.igem.org/Part:BBa_E1010">(E1010)</a></p> |
| + | </div> | ||
| + | |||
| + | <p>Fluorescent proteins are very common reporter genes. This particular one comes from <i>Discosoma striata</i> and was engineered to be monomeric. The excitation peak is at 584 nm, and the emission peak at 607 nm.</p> | ||
| + | |||
| + | <BR><BR> | ||
| + | <div class="section-title" style="text-align:center;float:left;width:100%;margin-bottom:0"> | ||
| + | <span><Marine style="margin-left:32px;font-family:'Brush Script MT';font-size:32px;text-transform:none;font-weight:bold;">Composite parts</Marine></span> | ||
| + | </div> | ||
| + | <br><br> | ||
| + | <div class="section-title" style="text-align:left;float:left;width:100%;margin-bottom:0"> | ||
| + | <p class="montserrat-text uppercase">piFinal <a style="color:#0066cc;" href="http://parts.igem.org/Part:BBa_K577882">(BBa_K577882)</a></p> | ||
</div> | </div> | ||
| − | <p> | + | <p>iGEM plasmid consisting of araC, a pBAD/AraC promoter (I0500), an RBS (B0034), an RFP (E1010), and two terminators (B0010 and B0012) contained inside one biobrick (B0015)</p> |
| + | <img src="https://static.igem.org/mediawiki/2017/3/34/UCLOUVAIN_-_BBa_K577882_approach_1.png" class="in_text_img" style="width: 500px;"> | ||
| − | |||
| − | + | </div> | |
| − | + | </div> | |
| − | + | ||
| − | + | ||
| − | + | ||
| − | |||
| − | |||
| − | |||
| − | <p>The | + | <div class="col-md-6"> |
| + | <div class="section-title" style="text-align:center;float:left;width:100%;margin-bottom:0"> | ||
| + | <span>COMR/S Approach</span> | ||
| + | <BR><BR> | ||
| + | <div class="section-title" style="text-align:center;float:left;width:100%;margin-bottom:0"> | ||
| + | <span><Marine style="margin-left:32px;font-family:'Brush Script MT';font-size:32px;text-transform:none;font-weight:bold;">Basic parts</Marine></span> | ||
| + | </div> | ||
| + | <BR><BR> | ||
| + | |||
| + | <div class="section-title" style="text-align:center;float:left;width:100%;margin-bottom:0"> | ||
| + | <p><center><i>New parts</i></center></p> | ||
| + | </div> | ||
| + | <BR><BR> | ||
| + | |||
| + | <div class="section-title" style="text-align:left;float:left;width:100%;margin-bottom:0"> | ||
| + | <p class="montserrat-text uppercase">ComR <a style="color:#0066cc;" href="http://parts.igem.org/Part:BBa_K2341000">(BBa_K2341000)</a></p> | ||
| + | </div> | ||
| + | |||
| + | <p>Protein originally found in <i>Streptococcus thermophilius</i> where it regulates natural competence. The protein activation and binding to specific promoters (ComR Box; BBa_K2341001) is only possible in presence of a short peptide, ComS (LPYFAGCL). Binding to the promoter will induce the downstream sequence expression.</p> | ||
<BR><BR> | <BR><BR> | ||
| − | + | <div class="section-title" style="text-align:left;float:left;width:100%;margin-bottom:0"> | |
| − | <p class="montserrat-text uppercase"> | + | <p class="montserrat-text uppercase">P1655 promoter <a style="color:#0066cc;" href="http://parts.igem.org/Part:BBa_K2341000">(BBa_K2341001)</a></p> |
| − | + | </div> | |
| − | <p> | + | <p>Promote expression of the sequence downstream in presence of ComR (BBa_K2341000) and ComS (LPYFAGCL). Sequence containing the palindrome motif needed to bind ComR (= ComR box).</p> |
<BR><BR> | <BR><BR> | ||
| − | + | <div class="section-title" style="text-align:left;float:left;width:100%;margin-bottom:0"> | |
| − | <p class="montserrat-text uppercase"> | + | <p class="montserrat-text uppercase">sgRNA Tyrosin (Cas9 guide) <a style="color:#0066cc;" href="http://parts.igem.org/Part:BBa_K2341100">(BBa_K2341100)</a></p> |
| − | + | </div> | |
| − | <p> | + | <p>RNA guide to use in combination with CRISPR/CAS9. Cut inside the tyrA gene so that deletion by homologous recombination can occur if a homologous DNA strand is also added. RNA guide under constitutive expression by BBa_J23119.</p> |
<BR><BR> | <BR><BR> | ||
| − | + | <div class="section-title" style="text-align:center;float:left;width:100%;margin-bottom:0"> | |
| − | + | <span><Marine style="margin-left:32px;font-family:'Brush Script MT';font-size:32px;text-transform:none;font-weight:bold;">Composite parts</Marine></span> | |
| − | < | + | </div> |
| + | <br><br> | ||
| + | <div class="section-title" style="text-align:left;float:left;width:100%;margin-bottom:0"> | ||
| + | <p class="montserrat-text uppercase">pCMMCD 105</p> | ||
</div> | </div> | ||
| − | <p> | + | <p>Plasmid consisting of ComR (BBa_K2341000), the p1655 promoter (containing the ComR box, BBa_K2341001), GusA and an erythromycin resistance.</p> |
| + | |||
| + | <p>GusA encodes the <i>beta</i>-glucuronidase gene from <i>E. coli</i>. This is a popular reporter gene that uses glucuronides as substrates, the most common of which are X-gluc (5-bromo-4-chloro-3-indolyl glucuronide) and <i>para</i>-nitrophenyl glucuronide.</p> | ||
| + | |||
| + | <p>Here, it is under the control of the ComRS pathway and uses the ComS peptide from S. thermophilius (LPYFAGGL).</p> | ||
<BR><BR> | <BR><BR> | ||
| + | <br><br> | ||
| + | </div> | ||
| + | <BR><br> | ||
| + | </div> | ||
</div> | </div> | ||
| − | |||
</section> | </section> | ||
| + | |||
</html> | </html> | ||
| + | <center><groupparts>iGEM17 UCLouvain</groupparts></center> | ||
{{UCLouvainFooter}} | {{UCLouvainFooter}} | ||
Latest revision as of 17:45, 1 November 2017
Parts
Auxotrophic Approach
pBAD/araC (I0500)
The pBAD promoter is a very common inducible one that is activated by L-arabinose and repressed by araC.
Terminator (B0010)
Terminators are necessary to stop the transcription at the end of a coding sequence.
RBS (B0034)
Efficient ribosome binding site characterized by IIT Madras in iGEM 2016.
RFP (E1010)
Fluorescent proteins are very common reporter genes. This particular one comes from Discosoma striata and was engineered to be monomeric. The excitation peak is at 584 nm, and the emission peak at 607 nm.
piFinal (BBa_K577882)
iGEM plasmid consisting of araC, a pBAD/AraC promoter (I0500), an RBS (B0034), an RFP (E1010), and two terminators (B0010 and B0012) contained inside one biobrick (B0015)
COMR/S Approach
Basic parts
New parts
Composite parts
ComR (BBa_K2341000)
Protein originally found in Streptococcus thermophilius where it regulates natural competence. The protein activation and binding to specific promoters (ComR Box; BBa_K2341001) is only possible in presence of a short peptide, ComS (LPYFAGCL). Binding to the promoter will induce the downstream sequence expression.
P1655 promoter (BBa_K2341001)
Promote expression of the sequence downstream in presence of ComR (BBa_K2341000) and ComS (LPYFAGCL). Sequence containing the palindrome motif needed to bind ComR (= ComR box).
sgRNA Tyrosin (Cas9 guide) (BBa_K2341100)
RNA guide to use in combination with CRISPR/CAS9. Cut inside the tyrA gene so that deletion by homologous recombination can occur if a homologous DNA strand is also added. RNA guide under constitutive expression by BBa_J23119.
pCMMCD 105
Plasmid consisting of ComR (BBa_K2341000), the p1655 promoter (containing the ComR box, BBa_K2341001), GusA and an erythromycin resistance.
GusA encodes the beta-glucuronidase gene from E. coli. This is a popular reporter gene that uses glucuronides as substrates, the most common of which are X-gluc (5-bromo-4-chloro-3-indolyl glucuronide) and para-nitrophenyl glucuronide.
Here, it is under the control of the ComRS pathway and uses the ComS peptide from S. thermophilius (LPYFAGGL).
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