Device

Part:BBa_K4062023

Designed by: Benjamin Bateman   Group: iGEM21_WLC-Milwaukee   (2021-10-16)


lactose regulated bla, lacI promoter inactivated

This part regulated b-lactamase expression using environmental lactose. The -10 sequence of the lacI promoter was deleted to negate the effects of the lac repressor. Six total bases were removed from the -10 sequence. The mutation was verified by sequencing.

Additionally, the mutation is verified by observing b-lactamases lack of interaction with its chromogenic substrate nitrocefin. Expression of b-lactamase is not expected in an environment with a high concentration of glucose and low concentration of lactose. However, as pictured below, that is what is observed. Red color of culture indicates expression of b-lactamase.


T--WLC-Milwaukee--lacIpromoter.png


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2913
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1981


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