Difference between revisions of "Part:BBa K2680060"

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===Mechanics===
 
===Mechanics===
  
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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.  
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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===
 
===Role in IFFL===
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"https://static.igem.org/mediawiki/2018/7/7b/T--William_and_Mary--034%2B032.png"
 
"https://static.igem.org/mediawiki/2018/7/7b/T--William_and_Mary--034%2B032.png"
  
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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 an incredibly weak promoter. This part could be applicable to teams who want to degrade a target protein, but want a low rate of degradation.
+
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.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 01:53, 18 October 2018


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

"T--William_and_Mary--034%2B032.png"

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 51
    Illegal NheI site found at 74
    Illegal NotI site found at 613
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 898
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE WITH RFC[1000]