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.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2913
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1981
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Categories
Parameters
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