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                     <p style="color:#FFFFFF;">The backbone of the U of Calgary project are the composite parts. Our composite parts are an amalgamation of different registry parts which act together to provide a novel function. The composite parts designed for this project are based around the modified Bowman-Birk Protease Inhibitor (mBBI) and the competency-inducing <a href="http://parts.igem.org/Part:BBa_K1444018">ComK part</a> that was submitted to the registry by the <a href="https://2014.igem.org/Team:Calgary/Notebook/Parts">2014 UofC_Calgary</a> team.</p><br>

<p style="color:#FFFFFF;">A variety of different parts were used across all of the mBBI composite constructs. We used Pveg, a constitutive <i>E. coli</i> and <i>B. subtilis</i> promoter, and double terminators to book-end internal genes upon chromosomal integration. To improve secretion and transdermal delivery of mBBI from <i>B. subtilis</i>, we added a secretion (sec) tag as well as a transdermal (TD1) tag. The secretion tag is a localization tag for the Sec pathway in <i>B. subtilis</i> (Ruan <i>et al.,</i> 2013). <a href="http://parts.igem.org/Part:BBa_E0040:Experience">TD1</a> is a part previously developed by the <a href="https://2013.igem.org/Team:USTC_CHINA">USTC_China 2013</a> iGEM team, and it is designed to allow for the trafficking of peptides across the skin.</p><br>
 
<p style="color:#FFFFFF;">The second major composite part was the ComK construct. ComK is a transcription factor and master regulator for competency in <i>B. subtilis</i>. We modified the original <a href="http://parts.igem.org/Part:BBa_K1444018">ComK</a> sequence, which was first submitted by the <a href="https://2014.igem.org/Team:Calgary/Notebook/Parts">2014 UofC_Calgary</a> team, to remove unwanted restriction sites from within the <i>comK</i> gene itself. The <i>comK</i> gene and xylose-inducible promoter were then flanked with sequences homologous to the <i>amyE</i> locus in <i>B. subtilis</i>, a non-essential gene responsible for the degradation of starch. These flanking sequences were added to allow for the integration of a controllable ComK into the <i>B. subtilis</i> genome. The integration of a xylose-inducible ComK allows for the transformation of <i>B. subtilis</i> to be more time-efficient and less resource-intensive. </p><br>

<p style="color:#FFFFFF;">Our last construct does not fall into either of the two previous categories. The construct is an integration cassette, which is a mix of both the mBBI and Comk constructs. Our cassette contains <i>thrC</i> integration sequences which flank the sec and TD1 tag. However, the mBBI sequence is replaced with the restriction sites for BamHI and XmaI restriction enzymes. Addition of the restriction sites allows for directional cloning of any desired insert, allowing <i>B. subtilis</i> to be used as a platform for future biotherapeutics, creating portable a mini-pharmacy.
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                                   <h3 class="human-practices-div-2">BBa_K2008001
 
				   </h3>
								   <p class="human-practices-div-2">BBI with N-terminal KSCI solubility tag fused to GFP for expression in <em>E. coli</em><br><a href="http://parts.igem.org/Part:BBa_K2008001">Registry Link</a>
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                                   <h3 class="human-practices-div-2">BBa_K2008002
									</h3>
								   <p class="human-practices-div-2">BBI with N-terminal sec and TD1 tags fused to GFP and under the pVeg promoter for expression in <em>B. subtilis</em><br><a href="http://parts.igem.org/Part:BBa_K2008002">Registry Link</a>
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                                   <h3 class="human-practices-div-2">BBa_K2008003 
									</h3>
								   <p class="human-practices-div-2">BBI with N-terminal KSCI solubility, sec, and TD1 tags fused to GFP and under the pVeg promoter for expression in <em>B. subtilis</em><br><a href="http://parts.igem.org/Part:BBa_K2008003">Registry Link</a>   
									</p>               
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                                   <h3 class="human-practices-div-2">BBa_K2008004
									</h3>
								   <p class="human-practices-div-2">BBI with N-terminal sec and TD1 tags under the pVeg promoter for expression in <em>B. subtilis</em><br><a href="http://parts.igem.org/Part:BBa_K2008004">Registry Link</a>
  
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                                   <h3 class="human-practices-div-2">BBa_K2008005
				   </h3>
								   <p class="human-practices-div-2">BBI with KSCI solubility, sec, and TD1 tags under the pVeg promoter for expression in <em>B. subtilis</em><br><a href="http://parts.igem.org/Part:BBa_K2008005">Registry Link</a>
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                                   <h3 class="human-practices-div-2">BBa_K2008006
 
									</h3>
								   <p class="human-practices-div-2">Improved version of <i>comK</i> gene with removal of internal restriction sites<br><a href="http://parts.igem.org/Part:BBa_K2008006">Registry Link</a>
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                                   <h3 class="human-practices-div-2">BBa_K2008007 
									</h3>
								   <p class="human-practices-div-2">Improved version of <i>comK</i> with addition of flanking regions of <i>amyE</i> homology <br><a href="http://parts.igem.org/Part:BBa_K2008007">Registry Link</a> 
									</p>               
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                                   <h3 class="human-practices-div-2">BBa_K2008008 
									</h3>
								   <p class="human-practices-div-2">Modular secretion platform containing sec and TD1 tags with flanking regions of <i>thrC</i> homology for chromosomal integration in <i>B. subtilis</i><br><a href="http://parts.igem.org/Part:BBa_K2008008">Registry Link</a>  
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