Composite

Part:BBa_K2680070

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)


37C Temperature Sensitive mScarlet-I pdt#3e


37C Temperature Sensitive mScarlet-I pdt#3e

Components: J23107 ts-CI + R0051 mScarlet-I pdt#3e

Mechanics

In this circuit, temperature-sensitive CI is expressed constitutively by J23107, a medium-weak constitutive promoter. However, the CI repressor protein denatures at 37C, losing its ability to bind the downstream CI promoter. Therefore, temperatures above 37C alleviate the repression of the CI promoter, allowing for free expression of the rapidly-maturing mScarlet-I. The mScarlet-I in this circuit has a pdt degradation tag, allowing for degradation by the mf-Lon protease. Specifically, this circuit contains pdt#3e, a very weak and slow degradation tag from part series K2333401-K2333406 or BBa_K2333406. To view a figure that displays the degradation rates of mScarlet-I with various pdt tags (relative to untagged mScarlet-I), see BBa_K2680251.

Role in IFFL

In this circuit, heat activates a reporter, mScarlet-I. In a complete IFFL (incoherent feed forward loop), an input must activate both a reporter and its inhibitor. Therefore, this circuit requires mf-Lon (or a different inhibitor of mScarlet-I) to act as an IFFL. Undef

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 1518
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None