Composite

Part:BBa_K2680058

Designed by: Stephanie Do, Xiangyi Fang, Ethan Jones, Jessica Laury, Adam Oliver, Lillian Parr, Chengwu Shen, Tinh Son, Julia Urban, Yashna Verma, Hanmi Zou   Group: iGEM18_William_and_Mary   (2018-10-06)


39C Temperature-Sensitive mf-lon

39C Temperature-Sensitive mf-lon. Components: J23107 TPL + ptlpA BCD8 mf-lon thrL.

Mechanics

J23107 (a medium-weak constitutive promoter) constitutively expresses TlpA39, a repressor protein that undergoes a conformational change at 39C. Due to this change, the repressor protein loses its ability to bind the downstream promoter, pTlpAr. Increasing temperature to 39C thus alleviates the repression on pTlpAr, allowing expression of mf-Lon protease.

Role in IFFL

In an IFFL (incoherent feed-forward loop), an input activates a reporter and its inhibitor. This circuit produces the mf-Lon protease, which can degrade and thus inhibit promoters like mScarlet-I. Thus, for use in an IFFL, this circuit must be combined with a reporter-containing circuit. A degradation tag, such as a pdt tag, should follow the reporter coding sequence.

Characterization


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]


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Categories
Parameters
None