Composite

Part:BBa_K539151

Designed by: Yang, Yi-Pei   Group: iGEM11_NCTU_Formosa   (2011-09-22)
Revision as of 09:29, 4 October 2011 by Bettyyang (Talk | contribs)

promoter+crtEBI+RNA thermometer(FourU)+crtY+terminator

This part is a generator of crtEBIY. Promoter BBa_J23103 + CrtEBI(BBa_K274100) is submitted by team Peking 2010. Enzyme cassette CrtEBI (with individual rbs) converts colourless farnesyl pyrophosphate to red lycopene (via intermediates geranylgeranyl pyroiphosphate and phytoene). And CrtY converting lycopene to orange beta-carotene. Our team assemble BBa_K346090(promoter+crtEBI) and RNA thermometer+crtY+terminator. This RNA thermometer initiate translation at 37°C. The existence of RNA thermometer makes crtY translated at specific temperature. We construct another circuit BBa_K539281 [1]included crtZ that can cooperate with circuit BBa_K539151. If E.coli is incubating below 37℃, mRNAs cannot be translated by ribosomes, because temperature-sensitive RBS(ribosome biding site) BBa_K115002 forms the hairpin. A ribosome cannot bind to the RBS so that it cannot translate CrtY BBa_K118008. As a result, the E.coli only can produce CrtE, CrtB and CrtI BBa_K346090 which convert colorless Farnesyl pyrophosphate to red Lycopene. Crt-1.png When the temperature is at 37℃ or higher, the hairpin is denatured so temperature-sensitive RBS BBa_K115002 is activated . CrtY BBa_K118008 is translated, which convert red Lycopene to orange beta-Carotene.

<img src="Crt-2.png"> Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2017
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1553
    Illegal NgoMIV site found at 1683
    Illegal NgoMIV site found at 4654
    Illegal AgeI site found at 768
  • 1000
    COMPATIBLE WITH RFC[1000]


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