Composite

Part:BBa_K4062027

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


lactose regulated bla, lacO inactivated

This device will express b-lactamase regardless of environmental lactose concentration. The lacI biobrick contains a mutation in lacO that changes the fifth and sixth bases from GT to AC. This mutation is shown to greatly reduce the ability of the lac repressor to bind lacO [1].

Since the lac repressor cannot bind lacO, expression of b-lactamase is expected regardless of lactose concentration. This is observed (pictured).

T--WLC-Milwaukee--lacO.png


[1]: Sarah Swerdlow and Roel Schaaper. "Mutagenesis in the lacI gene target of E. coli: improved analysis for lacI and lacO mutants." Mutat Res. 2014 December 1; 77: 79-84.

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 2919
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1987


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Categories
Parameters
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