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Revision as of 22:22, 17 October 2018
37C Temperature Sensitive mScarlet-I pdt#3
Temperature-Sensitive mScarlet-I pdt#3. Components: J23107 ts-CI + R0051 mScarlet-I pdt#3
Mechanics
In this circuit, temperature-sensitive CI is expressed constitutively 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 the rapidly-maturing mScarlet-I. The mScarlet-I in this circuit has a pdt, a protein degradation tag, allowing for degradation by mf-Lon protease (not included in this circuit). Specifically, this circuit contains pdt#3, a strong degradation tag from the part series K2333401-K2333406 or K2333401.
Role in an IFFL System
This circuit provides the reporter (mscarlet-I-pdt#3) for an IFFL system. Specifically, this reporter is able to be degraded by mf-Lon. When these two circuits are combined into a single cells by having them both be on different plasmids, IFFL like behavior should be possible. However, through experimentation, we have not identified an mf-Lon source strong enough to produce a pulse, that does not also kill the cells.
<img src="" width = "30%"/> </figure>
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 51
Illegal NheI site found at 74
Illegal NotI site found at 1518 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Unknown
- 1000COMPATIBLE WITH RFC[1000]