Difference between revisions of "Part:BBa K2680050"
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Mf-Lon is capable of degrading proteins tagged with a specific pdt tag. If used in combination with such a tagged protein, an IFFL circuit can be produced. During our research, we utilized BBa_K2680050 as our initial heat sensitive mf-Lon. After experimentation, we found that the amount of mf-Lon produced was too high to create an IFFL circuit, as either not enough mscarlet-I could be produced to generate a pulse, or the amount of mf-Lon and mscarlet-I was too high, causing cell death (see the figure below). | Mf-Lon is capable of degrading proteins tagged with a specific pdt tag. If used in combination with such a tagged protein, an IFFL circuit can be produced. During our research, we utilized BBa_K2680050 as our initial heat sensitive mf-Lon. After experimentation, we found that the amount of mf-Lon produced was too high to create an IFFL circuit, as either not enough mscarlet-I could be produced to generate a pulse, or the amount of mf-Lon and mscarlet-I was too high, causing cell death (see the figure below). | ||
| − | https://static.igem.org/mediawiki/2018/1/1f/T--William_and_Mary--Heat_Figure_2.png | + | "https://static.igem.org/mediawiki/2018/1/1f/T--William_and_Mary--Heat_Figure_2.png" |
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Revision as of 00:37, 18 October 2018
J23107 ts-CI+ R0051 mf-Lon
Design
This part is a j23107 ts-cI repressed mf-Lon. For more information on mf-Lon, and to see the source of the mf-Lon, see BBa_K2333011
Mechanics
In this circuit, temperature-sensitive CI is expressed by J23107, a medium-weak constitutive promoter. This temperature-sensitive CI then represses the downstream CI promoter. Once the temperature is sufficiently raised, the ts-CI protein denatures, loosing its repression capabilities. Therefore, the CI promoter is freed, allowing for expression of mf-Lon.
Characterization
Mf-Lon is capable of degrading proteins tagged with a specific pdt tag. If used in combination with such a tagged protein, an IFFL circuit can be produced. During our research, we utilized BBa_K2680050 as our initial heat sensitive mf-Lon. After experimentation, we found that the amount of mf-Lon produced was too high to create an IFFL circuit, as either not enough mscarlet-I could be produced to generate a pulse, or the amount of mf-Lon and mscarlet-I was too high, causing cell death (see the figure below).
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Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2903
Illegal AgeI site found at 2987
Illegal AgeI site found at 3193
Illegal AgeI site found at 3218 - 1000COMPATIBLE WITH RFC[1000]