Part:BBa_K2680060
39C Temperature-Sensitive (ptlpAr) mf-Lon
39C Temperature-Sensitive (ptlpAr) mf-lon. Components: J23107 TPL + ptlpAr BCD8 mf-lon thrL
Mechanics
In this circuit, a temperature-sensitive protein, TLP is expressed constitutively by J23107, a medium-weak constitutive promoter. When the temperature is increased to 39C, TLP looses its ability to repress a downstream weak pTLPAr promoter. Therefore, temperatures above 39C allow for the expression of mf-Lon.
Role in IFFL
This circuit prodes mf-Lon, a component that can be utilized to create an IFFL system. Mf-Lon is able to degrade proteins marked with special protein degradation tags. In an IFFL system, a reporter protein, such as mscarlet-I, can be tagged with a pdt tag. Experssion of mf-lon and mscarlet-I can then be initiated at the same time to produce a circuit with IFFL behavior. In our testing, we found that
Characterization
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As can be seen in the above figure, the amount of mf-Lon produced was not enough to sufficiently degrade ptlpA mscarlet-I to produce IFFL behavior. This is not surprising, as pTLPAr is a very weak promoter. This part could be applicable to teams who want to degrade a target protein, but want a low rate of degradation.
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 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 898
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1201
Illegal AgeI site found at 3305
Illegal AgeI site found at 3389
Illegal AgeI site found at 3595
Illegal AgeI site found at 3620 - 1000COMPATIBLE WITH RFC[1000]
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