Part:BBa_K2680062

39C Temperature-Sensitive mScarlet-I pdt#3

39C Temperature-Sensitive mScarlet-I pdt#3. Components: J23107 TPL + ptlpA b0034m mScarlet-I pdt#3

Mechanics

J23107, a medium-weak constitutive promoter, constitutively expresses TlpA₃₉. Unlike the original TlpA, TlpA₃₉ undergoes a conformational change at 39C and loses its ability to bind the downstream pTlpA. Therefore, TlpA₃₉ halts its repression of pTlpA at temperatures above 39C, inducing the 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#3, the strongest degradation tag from part series K2333401-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.

As an 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.

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
  Illegal NotI site found at 1920
  
• 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
  
• 1000
  COMPATIBLE WITH RFC[1000]