Difference between revisions of "Team:Toronto/Parts"
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| − | + | <h1>Parts</h1> | |
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</div> | </div> | ||
| + | |||
| + | <!-- Content div --> | ||
| + | <div class="section bg-white"> | ||
| + | <div class="container content-page row"> | ||
| + | <div class="block content" style="max-width: 1000px;"> | ||
| + | |||
| + | <!-- subsection 1 --> | ||
| + | <div id="subsection-submitted-parts" class="subsection"> | ||
| + | |||
| + | <h2 class="text-red">Submitted Parts</h2> | ||
| + | |||
| + | <!-- Figure start --> | ||
| + | <figure> | ||
| + | <div class="figures"> | ||
| + | <table> | ||
| + | <thead> | ||
| + | <tr> | ||
| + | <th>pSB1C3</th> | ||
| + | <th>Registry number </th> | ||
| + | <th>Type</th> | ||
| + | <th>Length (bp)</th> | ||
| + | <th>Function </th> | ||
| + | <th>Assembly Method</th> | ||
| + | <th>UNS</th> | ||
| + | </tr> | ||
| + | </thead> | ||
| + | <tbody class="table-hover"> | ||
| + | <tr> | ||
| + | <td>LacILOV-mCherry </td> | ||
| + | <td><a style="text-decoration: underline !important" href="http://parts.igem.org/Part:BBa_K2469003">BBa_K2469003</a></td> | ||
| + | <td>Composite </td> | ||
| + | <td>1615</td> | ||
| + | <td>Light regulated reporter </td> | ||
| + | <td>Digestion and ligation </td> | ||
| + | <td>2, 3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>HIS-AcrIIA4</td> | ||
| + | <td><a style="text-decoration: underline !important" href="http://parts.igem.org/Part:BBa_K2469002">BBa_K2469002</a></td> | ||
| + | <td>Basic</td> | ||
| + | <td>398</td> | ||
| + | <td>Inhibits the function of S. pyogenes Cas9 and dCas9.</td> | ||
| + | <td>Gibson assembly of amplified AcrIIA4 from G-block and amplified pSB1C3 backbone from LacILOV-mCherry Construct in pSB1C3</td> | ||
| + | <td>2, 3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>LacILOV</td> | ||
| + | <td><a style="text-decoration: underline !important" href="http://parts.igem.org/Part:BBa_K2469001">BBa_K2469001</a></td> | ||
| + | <td>Basic</td> | ||
| + | <td>728</td> | ||
| + | <td>A fusion protein of the light, oxygen and voltage-sensitive domain of the Avena Sativa phototropin 1 (LOV2) and the LacI domain. This transcriptional regulator represses promoters under control of LacI. It is speculated that in the presence of blue light, LacILOV dissociates, causing it to unbind from DNA and allow transcription. In the absence of light, the protein dimerizes and binds to the promoter of lac operon, inhibiting the transcription of downstream genes.</td> | ||
| + | <td>Gibson assembly of amplified LacILOV part from LacILOV-mCherry construct in pKDL071 and pSB1C3 backbone from LacILOV-mCherrt construct in pSCB1C3</td> | ||
| + | <td>2, 3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>reporter construct </td> | ||
| + | <td><a style="text-decoration: underline !important" href="http://parts.igem.org/Part:BBa_K2469004">BBa_K2469004</a></td> | ||
| + | <td>Composite </td> | ||
| + | <td>3253</td> | ||
| + | <td>Light regulated reporter switch, alternating between mCherry and YFP flourescence </td> | ||
| + | <td>Gibson assembly of amplified reporter construct out of pkDL071 and amplified pSB1C3 backbone from the LacILOV-mCherry construct in pSB1C3 </td> | ||
| + | <td>2, 3</td> | ||
| + | </tr> | ||
| + | </tbody> | ||
| + | </table> | ||
| + | </div> | ||
| + | <figcaption>Table 1: List of parts that was submitted to iGEM registry.</figcaption> | ||
| + | </figure> | ||
| + | <!-- figure end --> | ||
| + | |||
| + | </div> | ||
| + | <!-- subsection 1 END --> | ||
| + | |||
| + | |||
| + | <!-- subsection 2 --> | ||
| + | <div id="subsection-other-parts" class="subsection"> | ||
| + | |||
| + | <h2 class="text-red">Other Parts</h2> | ||
| + | |||
| + | <!-- Figure start --> | ||
| + | <figure> | ||
| + | <div class="figures"> | ||
| + | <table> | ||
| + | <thead> | ||
| + | <tr> | ||
| + | <th>pKDLO71</th> | ||
| + | <th>Type </th> | ||
| + | <th>Length</th> | ||
| + | <th>Function</th> | ||
| + | <th>Assembly Method</th> | ||
| + | <th>UNS</th> | ||
| + | </tr> | ||
| + | </thead> | ||
| + | <tbody class="table-hover"> | ||
| + | <tr> | ||
| + | <td>LacILOV-mCherry</td> | ||
| + | <td>Composite </td> | ||
| + | <td>1615</td> | ||
| + | <td>Light regulated reporter </td> | ||
| + | <td>Gibson g-Block + Backbone with UNS</td> | ||
| + | <td>2, 3</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>reporter mCherry</td> | ||
| + | <td></td> | ||
| + | <td></td> | ||
| + | <td>reporter construct, consisting of the CI repressor (with LVA degradation tag) and mCherry under the control of the Ptrc-2 promoter; used as a negative control in the sgRNA assay </td> | ||
| + | <td>Gibson g-Block + Backbone with UNS</td> | ||
| + | <td>2, 4</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>reporter mCherry + sgRNA </td> | ||
| + | <td></td> | ||
| + | <td></td> | ||
| + | <td>Half of the full construct, to test the function of the sgRNA (ensure it is targetting the correct sequence) </td> | ||
| + | <td>Gibson assembly of sgRNA g-Block into reporter mCherry, with UNS 5 extension on backbone </td> | ||
| + | <td>4, 5</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>reporter mCherry + sgRNA+ AcrIIA4 </td> | ||
| + | <td></td> | ||
| + | <td></td> | ||
| + | <td>Incorporation of the Anti-CRISPR protein into the sgRNA construct to have both expressed simultaneously and test the function of the Anti-CRISPR </td> | ||
| + | <td>Gibson assemly of AcrIIA4 g-block into reporter mCherry + sgRNA, with UNS 6 extension on backbone </td> | ||
| + | <td>4, 6</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>reporter construct </td> | ||
| + | <td>Composite </td> | ||
| + | <td>3253</td> | ||
| + | <td>Light regulated reporter switch, alternating between mCherry and YFP flourescence </td> | ||
| + | <td>Gibson assembly of reporter YFP (LacILOV + YFP) into reporter mCherry with UNS 3 extension on backbone </td> | ||
| + | <td>3, 2</td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>full construct </td> | ||
| + | <td></td> | ||
| + | <td></td> | ||
| + | <td>Light-regulated switch controlling the activity of the CRISPR-Cas9 system. </td> | ||
| + | <td>Gibson assembly of AcrIIA4 subpart (LacILOV + AcrIIA4 + YFP) into reporter mCherry + sgRNA, with UNS 3 extension on bacbone </td> | ||
| + | <td>3, 6</td> | ||
| + | </tr> | ||
| + | </tbody> | ||
| + | </table> | ||
| + | </div> | ||
| + | <figcaption>Table 2: Other parts used in our project that was not submitted to iGEM registry.</figcaption> | ||
| + | </figure> | ||
| + | <!-- figure end --> | ||
| + | |||
| + | </div> | ||
| + | <!-- subsection 2 END --> | ||
| + | |||
| + | |||
| + | </div> | ||
| + | |||
| + | |||
| + | </div> | ||
</div> | </div> | ||
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{{Toronto/footer}} | {{Toronto/footer}} | ||
Latest revision as of 19:53, 14 December 2017
Parts
Submitted Parts
| pSB1C3 | Registry number | Type | Length (bp) | Function | Assembly Method | UNS |
|---|---|---|---|---|---|---|
| LacILOV-mCherry | BBa_K2469003 | Composite | 1615 | Light regulated reporter | Digestion and ligation | 2, 3 |
| HIS-AcrIIA4 | BBa_K2469002 | Basic | 398 | Inhibits the function of S. pyogenes Cas9 and dCas9. | Gibson assembly of amplified AcrIIA4 from G-block and amplified pSB1C3 backbone from LacILOV-mCherry Construct in pSB1C3 | 2, 3 |
| LacILOV | BBa_K2469001 | Basic | 728 | A fusion protein of the light, oxygen and voltage-sensitive domain of the Avena Sativa phototropin 1 (LOV2) and the LacI domain. This transcriptional regulator represses promoters under control of LacI. It is speculated that in the presence of blue light, LacILOV dissociates, causing it to unbind from DNA and allow transcription. In the absence of light, the protein dimerizes and binds to the promoter of lac operon, inhibiting the transcription of downstream genes. | Gibson assembly of amplified LacILOV part from LacILOV-mCherry construct in pKDL071 and pSB1C3 backbone from LacILOV-mCherrt construct in pSCB1C3 | 2, 3 |
| reporter construct | BBa_K2469004 | Composite | 3253 | Light regulated reporter switch, alternating between mCherry and YFP flourescence | Gibson assembly of amplified reporter construct out of pkDL071 and amplified pSB1C3 backbone from the LacILOV-mCherry construct in pSB1C3 | 2, 3 |
Other Parts
| pKDLO71 | Type | Length | Function | Assembly Method | UNS |
|---|---|---|---|---|---|
| LacILOV-mCherry | Composite | 1615 | Light regulated reporter | Gibson g-Block + Backbone with UNS | 2, 3 |
| reporter mCherry | reporter construct, consisting of the CI repressor (with LVA degradation tag) and mCherry under the control of the Ptrc-2 promoter; used as a negative control in the sgRNA assay | Gibson g-Block + Backbone with UNS | 2, 4 | ||
| reporter mCherry + sgRNA | Half of the full construct, to test the function of the sgRNA (ensure it is targetting the correct sequence) | Gibson assembly of sgRNA g-Block into reporter mCherry, with UNS 5 extension on backbone | 4, 5 | ||
| reporter mCherry + sgRNA+ AcrIIA4 | Incorporation of the Anti-CRISPR protein into the sgRNA construct to have both expressed simultaneously and test the function of the Anti-CRISPR | Gibson assemly of AcrIIA4 g-block into reporter mCherry + sgRNA, with UNS 6 extension on backbone | 4, 6 | ||
| reporter construct | Composite | 3253 | Light regulated reporter switch, alternating between mCherry and YFP flourescence | Gibson assembly of reporter YFP (LacILOV + YFP) into reporter mCherry with UNS 3 extension on backbone | 3, 2 |
| full construct | Light-regulated switch controlling the activity of the CRISPR-Cas9 system. | Gibson assembly of AcrIIA4 subpart (LacILOV + AcrIIA4 + YFP) into reporter mCherry + sgRNA, with UNS 3 extension on bacbone | 3, 6 |
