37C Temperature Sensitive mScarlet-I pdt#3
Temperature-Sensitive mScarlet-I pdt#3. Components: J23107 ts-CI + R0051 mScarlet-I pdt#3
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, the strongest degradation tag from the part series K2333401-K2333406. To view a figure that displays the degradation rates of mScarlet-I with various pdt tags (relative to untagged mScarlet-I), see BBa_K2680251.
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.
For example, as can be seen in the above graph, when we try to combine a ts-CI mf-Lon on a psb3K3 plasmid with BBa_K2680051, the cells are killed upon dilution, and when the temperature is raised to 37C (grey region).
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 51
Illegal NheI site found at 74
Illegal NotI site found at 1518
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC