Composite

Part:BBa_K5178016

Designed by: Virginia iGEM 2024   Group: iGEM24_Virginia   (2024-10-02)


RAT_sequence_lac o_T7RNAP

This composite part is composed of the pPtsG promoter, a lac operator (lac o) sequence, the terminator/anti-terminator sequence, the coding sequence for T7 RNA polymerase, and the B0015 terminator. The terminator/anti-terminator sequence is composed of an RNA anti-terminator (RAT) sequence. When the RAT sequence is unbound, a terminator hairpin forms in the RNA, preventing further transcription. When the RAT sequence is bound (with dephosphorylated GlcT, which is glucose-dependent), the terminator hairpin is suppressed, and transcription can proceed. Additionally, the lac o sequence allows for binding by the lac repressor protein, LacI. The lac o regulates downstream expression of genes of interest based on LacI presence. This way, the expression of T7RNAP can be modulated by glucose/LacI presence.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2972
    Illegal NheI site found at 2976
    Illegal NheI site found at 2980
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2468
    Illegal XhoI site found at 787
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2901
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1390


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Parameters
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