Plasmid

Part:BBa_K5101005

Designed by: Ziqi Mi   Group: iGEM24_ZQT-Nanjing   (2024-09-11)
Revision as of 13:12, 11 September 2024 by Registry (Talk | contribs)

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pET29(a)-pJ23119-TIpA39-T-pTIpA-RBS-PhiX174E-T7

To enable the engineered bacteria to suicide when released into the environment (generally above 39 degrees Celsius) during fecal disposal, we chose a temperature-responsive protein TlpA39(BBa_K2572000) and its corresponding promoter pTlpA(BBa_K2500003) as regulatory elements to induce the expression of the lysis protein PhiX174E. TlpA39 is derived from Salmonella enterica. It is a temperature-sensitive protein that can bind to pTlpA. At temperatures above 39 degrees Celsius, the TlpA39 protein will remain in a monomeric form, thereby losing the ability to bind to the promoter pTlpA. If the temperature drops below 39 degrees Celsius, the TlpA39 protein will form a dimer; the dimer can inhibit the pTlpA and prevent the recruitment of RNA polymerase, thereby suppressing downstream gene expression.

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
    Illegal NheI site found at 1205
    Illegal NotI site found at 572
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 857
    Illegal XhoI site found at 1632
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1160
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None