Regulatory

Part:BBa_K833005

Designed by: Julianne Grose   Group: iGEM12_BYUProvo   (2012-10-02)

Mutated prfA-UTR thermosensor from Listeria monocytogenes


This part was derived from part BBa_K619889 via TAQ random mutagenesis and screening for thermosensors that responded to a 35 to 37'C shift. In contrast to the our wild type thermosensor (part BBa_K619889) which activates translation from a 30 to 37 degree shift, this thermosensor activates transcription from a 35 to 37 degree shift (a more concise dynamic range). This mutation was isolated by randomly mutagenizing part BBa_K619889 and monitoring a lacZ output. The thermosensor has a shine delgarno and initiating ATG that are barried within a stem-loop structure at 30'C, but are accessible at higher temperatures as the mRNA structure is disrupted. This thermosensor is part of a library of thermosensors from this screen (see parts (Bba_K833000 to Bba_K833010 and also BBa_K619890,BBa_K61989 -619894 from our 2011 submission). mutated prfA thermosensor from listeria D: This is the prfA-UTR from Listeria monocytogenes which has been randomly mutated to produce a thermosensor this a more discrete dynamic range.

To use: fuse a reporter (such as LacZ, GFP or RFP) to the initiating ATG in frame

Plate assay: In contrast to the our wild type thermosensor (part BBa_K619889) which activates translation from a 30 to 37 degree shift, this thermosensor activates transcription from a 35 to 37 degree shift. This phenotype has been confirmed 3 times on X-gal plates using the LacZ reporter.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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