Composite

Part:BBa_K2353002

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)
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tsPurpleLAA

tsPurpleLAA is composed of a ribosomal binding site, tsPurple (a purple chromoprotein which functions as a reporter), and LAA (a fast degradation tag). tsPurpleLAA is intended to be assembled with pLambdaR-LacI (which is the promoter and regulatory system) and r0011ClpXPCI (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 tsPurpleLAA 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 LAA deg tag and degrade the tsPurple protein. Since LAA is a fast degradation tag, so tsPurpleLAA should have the highest levels of degradation seen. 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]


[edit]
Categories
Parameters
None