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Part:BBa_K3985025

Designed by: Andrea Aquize, Martina Buerkle, Maira Bradler, Sidney Mitsch, Emilia Marie Wrede   Group: iGEM21_TU_Darmstadt   (2021-10-16)


lox66 fixed

lox66 is the right element (RE) loxP mutant if annotated in the direction right to left.[3] When used in combination with lox71 (left element (LE) loxP mutant with the same annotation as above) Cre-Lox recombination leads to the formation of a double-mutant lox72 site, having a reduced affinity for Cre recombinase. Hence, repeated Cre-Lox recombination becomes very unlikely. Thus, the recombination event is regarded as "irreversible".

We use lox66 to perform Cre-Lox inversion of our synthetic kill-switch cassette. Hence, lox66 changes to wild-type loxP. As we combine lox66 with lox71, the Cre-Lox recombination thus turns irreversible due to the resulting double-mutant lox72 site.

This part is an improvement of the existing registry part: BBa_I718016 where the sequence for lox66 is incorrect.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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