Line 30: | Line 30: | ||
<h2>Submitted Parts</h2> | <h2>Submitted Parts</h2> | ||
+ | |||
+ | <figure> | ||
+ | <div class="figures"> | ||
+ | <table> | ||
+ | <thead> | ||
+ | <tr> | ||
+ | <th>Time</th> | ||
+ | <th>Quantity</th> | ||
+ | <th>iunno</th> | ||
+ | </tr> | ||
+ | </thead> | ||
+ | <tbody class="table-hover"> | ||
+ | <tr> | ||
+ | <td>Hello</td> | ||
+ | <td>9adsf asd</td> | ||
+ | <td>2 ads fads fdsa s </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>2 ads fads fdsa s </td> | ||
+ | <td>2 ads fads fdsa s </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>2</td> | ||
+ | <td>2 ads fads fdsa s </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>2</td> | ||
+ | <td>2 ads fads fdsa s </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Hello</td> | ||
+ | <td>911 asd afd </td> | ||
+ | <td>2 ads fads fdsa s </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>2</td> | ||
+ | <td>2 ads fads fdsa s </td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | </div> | ||
+ | <figcaption>Figure 1.1: Wildlife population is wildlife population is wildlife population is wildlife population</figcaption> | ||
+ | </figure> | ||
+ | |||
<!-- Figure start --> | <!-- Figure start --> |
Revision as of 00:48, 1 November 2017
Parts
Submitted Parts
Time | Quantity | iunno |
---|---|---|
Hello | 9adsf asd | 2 ads fads fdsa s |
1 | 2 ads fads fdsa s | 2 ads fads fdsa s |
1 | 2 | 2 ads fads fdsa s |
1 | 2 | 2 ads fads fdsa s |
Hello | 911 asd afd | 2 ads fads fdsa s |
1 | 2 | 2 ads fads fdsa s |
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. In the presence of blue light this protein will act as a repressor, and in its absence the protein will dissociate from the promoter | 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 |
References
- Canadian Biosafety Standards and Guidelines - https://www.canada.ca/en/public-health/services/canadian-biosafety-standards-guidelines.html
- Biosafety Team in the Office of Environmental Health and Safety - https://ehs.utoronto.ca/our-services/biosafety/
- NIH Guidelines - https://www.canada.ca/en/public-health/services/laboratory-biosafety-biosecurity/pathogen-safety-data-sheets-risk-assessment.html
- iGEM Risk Groups - https://2017.igem.org/Safety/Risk_Groups