Difference between revisions of "Part:BBa K2680062"
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===Mechanics=== | ===Mechanics=== | ||
− | J23107, a medium-weak constitutive promoter, constitutively expresses TlpA<sub>39</sub>. Unlike the original TlpA, TlpA<sub>39</sub> undergoes a conformational change at 39C and loses its ability to bind the downstream pTlpA. Therefore, TlpA<sub>39</sub> halts its repression of pTlpA at temperatures above 39C, inducing the 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 K2333401-K2333406 | + | J23107, a medium-weak constitutive promoter, constitutively expresses TlpA<sub>39</sub>. Unlike the original TlpA, TlpA<sub>39</sub> undergoes a conformational change at 39C and loses its ability to bind the downstream pTlpA. Therefore, TlpA<sub>39</sub> halts its repression of pTlpA at temperatures above 39C, inducing the 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 K2333401-K2333406. 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>. |
===As an IFFL=== | ===As an IFFL=== |
Revision as of 02:54, 18 October 2018
39C Temperature-Sensitive mScarlet-I pdt#3
39C Temperature-Sensitive mScarlet-I pdt#3. Components: J23107 TPL + ptlpA b0034m mScarlet-I pdt#3
Mechanics
J23107, a medium-weak constitutive promoter, constitutively expresses TlpA39. Unlike the original TlpA, TlpA39 undergoes a conformational change at 39C and loses its ability to bind the downstream pTlpA. Therefore, TlpA39 halts its repression of pTlpA at temperatures above 39C, inducing the 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 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.
As an 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.
Characterization
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 613
Illegal NotI site found at 1920 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 898
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1201
- 1000COMPATIBLE WITH RFC[1000]