Construction and Testing
From 2007.igem.org
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03873 BBa_S03873] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03873 BBa_S03873] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_1.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_1.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03877 BBa_S03877]</td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03877 BBa_S03877]</td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_3.png]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_3.png]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03878 BBa_S03878] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03878 BBa_S03878] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_4.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_4.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> LasI protein generator </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03882 BBa_S03882] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03882 BBa_S03882] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_14.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_14.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> LasR protein generator </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03874 BBa_S03874] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03874 BBa_S03874] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_12.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_12.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> </td> |
</tr> | </tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03875 BBa_S03875] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03875 BBa_S03875] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_2.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_2.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> PcyA protein generator </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03879 BBa_S03879] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03879 BBa_S03879] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_13.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_13.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> </td> |
</tr> | </tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03880 BBa_S03880] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03880 BBa_S03880] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_5.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_5.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> TetR protein generator </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03885 BBa_S03885] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03885 BBa_S03885] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_8.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_8.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> functionality unconfirmed</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03883 BBa_S03883] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03883 BBa_S03883] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_7.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_7.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> </td> |
</tr> | </tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03887 BBa_S03887] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03887 BBa_S03887] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_9.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_9.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> functionality confirmed </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03888 BBa_S03888] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03888 BBa_S03888] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_10.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_10.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> functionality confirmed </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03889 BBa_S03889] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03889 BBa_S03889] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_11.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_11.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> functionality confirmed </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03881 BBa_S03881] </td> | <td style="border-bottom:1px solid black;padding:5px"> [http://partsregistry.org/Part:BBa_S03881 BBa_S03881] </td> | ||
<td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_6.PNG]] </td> | <td style="border-bottom:1px solid black;padding:5px"> [[Image:UW_6.PNG]] </td> | ||
- | <td style="border-bottom:1px solid black;padding:5px"> | + | <td style="border-bottom:1px solid black;padding:5px"> functionality confirmed </td> |
</tr> | </tr> | ||
</table> | </table> |
Revision as of 00:40, 27 October 2007
Contents |
Construction Tree
The diagram below outlines our parallel construction plan, from individual registry parts to the final half-adder. Because many people were working on the project at once, it is designed to keep everyone organised.
Diagram details:
- Expected fragment sizes are shown at every node so gel results can be analyzed quickly.
- Vector resistances for each part/construct are shown, with blue representing ampicillin resistance and red representing kanamycin resistance. Purple represents a resistance to both.
- Parts and developing constructs all have abbreviated names for ease of labeling. To make the subtle distinction between a coding sequence and a coding sequence with a ribosome binding site, we use a ' on the computer and an underline when writing. A double ' or double underline indicates a coding sequence with a ribosome binding site and a transcriptional terminator.
- Arrows show the direction of DNA cloning and specify which restriction enzymes to use for cutting. Hollow arrows are present when two parts are ligated from vectors of the same resistance, requiring additional methods such as the use of alkaline phosphatase, gel extraction, or a three-way ligation into a vector with different resistance.
- The numbers 1-2-3-3.5-4 indicate which major protocols need to be carried at each step in the assembly cycle.
Testing Constructs
We use the fluorescent proteins GFP and RFP as reporter genes to test the functionality of some of the components and constructs of our half-adder design. Where possible, we make use of the lac inducible promoter to control expression of the enhancing or repressing elements.
Comparing basal and induced expression levels for the quorum sensing promoters
Testing functionality of the various promoters
Submitted Parts to the Registry
The following is a table listing all of the parts submitted to the registry.
Name | Part Number | Constructs | Comments |
rfp' | BBa_S03873 | ||
ho1' | BBa_S03877 | ||
lasi | BBa_S03878 | LasI protein generator | |
lasr | BBa_S03882 | LasR protein generator | |
pcyA' | BBa_S03874 | ||
pcya | BBa_S03875 | PcyA protein generator | |
tetr' | BBa_S03879 | ||
tetr | BBa_S03880 | TetR protein generator | |
plas rfp' | BBa_S03885 | functionality unconfirmed | |
plac lasr | BBa_S03883 | ||
ptet rfp' | BBa_S03887 | functionality confirmed | |
pomp rfp' | BBa_S03888 | functionality confirmed | |
plux rfp' | BBa_S03889 | functionality confirmed | |
plasoci rfp' | BBa_S03881 | functionality confirmed |
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