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 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 degrade a target protein, but want a low rate of degradation.
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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.
  
 
<!-- Add more about the biology of this part here
 
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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]