Difference between revisions of "Part:BBa K2680053"
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Tight Repression 37C Temperature-Sensitive mScarlet-I pdt#3. | Tight Repression 37C Temperature-Sensitive mScarlet-I pdt#3. | ||
Components: J23100 ts-CI + R0051 mScarlet-I pdt#3 | Components: J23100 ts-CI + R0051 mScarlet-I pdt#3 | ||
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===Mechanics=== | ===Mechanics=== | ||
| − | In this circuit, temperature-sensitive CI is expressed constitutively by J23100, a strong constitutive promoter. However, the CI repressor protein denatures at 37C, losing its ability to bind the downstream CI promoter. Therefore, temperatures above 37C alleviate the repression of the CI promoter, allowing for free expression of the rapidly-maturing mScarlet-I. The mScarlet-I in this circuit has a pdt degradation tag, allowing for degradation by the mf-Lon protease. Specifically, this circuit contains pdt#3, | + | In this circuit, temperature-sensitive CI is expressed constitutively by J23100, a strong constitutive promoter. However, the CI repressor protein denatures at 37C, losing its ability to bind the downstream CI promoter. Therefore, temperatures above 37C alleviate the repression of the CI promoter, allowing for free expression of the rapidly-maturing mScarlet-I. The mScarlet-I in this circuit has a pdt degradation tag, allowing for degradation by the mf-Lon protease. Specifically, this circuit contains pdt#3, the strongest degradation tag from part series BBa_K2333001-Bba_K2333006. To view a figure that displays the degradation rates of mScarlet-I with various pdt tags (relative to untagged mScarlet-I), see <partinfo>BBa_K2680251</partinfo>. |
===Role in IFFL=== | ===Role in IFFL=== | ||
Latest revision as of 02:52, 18 October 2018
Tight Repression 37C Temperature-Sensitive mScarlet-I pdt#3
Tight Repression 37C Temperature-Sensitive mScarlet-I pdt#3. Components: J23100 ts-CI + R0051 mScarlet-I pdt#3
Mechanics
In this circuit, temperature-sensitive CI is expressed constitutively by J23100, a strong constitutive promoter. However, the CI repressor protein denatures at 37C, losing its ability to bind the downstream CI promoter. Therefore, temperatures above 37C alleviate the repression of the CI promoter, allowing for free expression of the rapidly-maturing mScarlet-I. The mScarlet-I in this circuit has a pdt degradation tag, allowing for degradation by the mf-Lon protease. Specifically, this circuit contains pdt#3, the strongest degradation tag from part series BBa_K2333001-Bba_K2333006. 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 IFFL
In this circuit, heat activates a reporter, mScarlet-I. In a complete IFFL (incoherent feed forward loop), an input must activate both a reporter and its inhibitor. Therefore, this circuit requires mf-Lon (or a different inhibitor of mScarlet-I) to act as an IFFL.
As can be seen in the above figure, when combined with a J23100 ts-cI repressed mf-Lon, the amount of mf-Lon produced, relative to the amount of m-scarlet produced, is too high, so no IFFL like pulse is observed.
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]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]