Composite

Part:BBa_K2353001

Designed by: Julia Leveille, Lauren Hong, Emily Gibson, Gaurav Byagathvalli, Katie Barr, Alyssa Franklin, Christina Lee, Ellie Kim, Kevin Li, Megan Hong, Natalie Shih, Nithik Balachandran, Noora Chandasir, David S   Group: iGEM17_Lambert_GA   (2017-10-17)


tsPurpleDAS

tsPurpleDAS is composed of a ribosomal binding site, tsPurple (a purple chromoprotein which functions as a reporter), and DAS (a moderately fast degradation tag). tsPurpleDAS is intended to be assembled with pLambdaR-LacI (BBa_K1911000),which is the promoter and regulatory system, and pLac-ClpXP-CI (BBa_K1911001),which is the non-lysosomal protease. The terminator is located at the beginning of pLac-ClpXP-CI. When fully assembled, p-lambda-r LacI transcribes tsPurpleDAS and LacI represses pLac, preventing transcription of ClpXP-CI. Upon induction with IPTG, LacI binds to IPTG which prevents repression of pLac. Therefore, ClpXP-CI is transcribed; ClpXP should recognize the DAS deg tag and degrade the tsPurple protein. The ensuing levels of expression could then be compared to that of BBa_K2353000 (tsPurple without a deg tag) to see the relative level of degradation experienced.

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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Categories
Parameters
None