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. 2016 Dec 13;6:38952. doi: 10.1038/srep38952

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Copyright © 2016, The Author(s)

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(A) After transformation the linear GFP expression cassette from pAHBgl-GFP moved to chromosome 4, where the AOX1 locus is situated. (B) Two distal homologous sequences of the cassette with the chromosomal AOX1 locus mediate a double cross-over recombination event (C) The cassette has replaced the AOX1 locus on chromosome 4 (D) The excised linear AOX1 locus moved to chromosome 2 (E) Potentially due to a DSB initiating the NHEJ pathway, the AOX1 locus is integrated into a random gene on chromosome 2 (F) The untargeted gene PP7435_Chr2-1130 was disrupted. While the AOX1 locus is still functional, the disruption of the untargeted gene likely led to the observed abnormal colony morphology in the case of JPS014.