Part:BBa_K5159015
Y. lipolytica KU70 3' homology arm
The sequence is found in the Yarrowia lipolytica genome and used for the knockout of KU70 gene by homologous recombination (HR). It is located downstream of the genomic KU70 coding region to be knocked out. This homologous arm, together with its upstream counterpart (BBa_K5159014) flanking a selection marker (e.g., LEU2 expression cassette), can locate the KU70 gene in the genome of Yarrowia lipolytica.
As an unconventional yeast species, Y. lipolytica prefers non-homologous end-joining (NHEJ) as the main mechanism for genomic integration instead of homologous recombination, which generally occurs at a very low frequency. KU70 encodes a protein in charge of the double-strand break (DSB) repair in Yarrowia lipolytica enabling NHEJ. Knocking out KU70 can increase the chance for subsequent HR by several folds. As further knockouts via HR are involved, for example, knocking out PEX10 in our project, constructing a KU70 knockout strains would significantly increase the success rate of engineering.
A 1000 bp homologous arm in length is necessary for the efficient knockout to occur.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 304
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 304
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 304
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 304
- 1000COMPATIBLE WITH RFC[1000]
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