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 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 mscarlet to produce IFFL behavior. This is not surprising, as pTLPAr is an incredibly weak promoter. This part could be applicable to teams who want to add 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]
None |