CPC C12N 15/902 (2013.01) [C12N 5/0606 (2013.01); C12N 5/0647 (2013.01); C12N 5/0696 (2013.01); C12N 9/22 (2013.01); C12N 15/102 (2013.01); C12N 15/113 (2013.01); C12N 15/86 (2013.01); C12N 2310/20 (2017.05)] | 12 Claims |
1. An isolated nucleic acid for homologous recombination with a target nucleic acid having a cleavage site, wherein:
(a) a first strand of the target nucleic acid comprises, from 5′ to 3′, P1-H1-X-H2-P2, wherein
P1 is a first priming site;
H1 is a first homology arm;
X is the cleavage site;
H2 is a second homology arm; and
P2 is a second priming site; and
(b) a first strand of the isolated nucleic acid comprises, from 5′ to 3′, A1-S1-P2′-N-P1′-S2-A2, wherein
A1 is a homology arm that is substantially identical to H1;
P2′ is a priming site that is substantially identical to P2;
N is a cargo;
P1′ is a priming site that is substantially identical to P1;
A2 is a homology arm that is substantially identical to H2;
wherein S1 is a first stuffer, wherein S2 is a second stuffer, and wherein each of S1 and S2 comprise a random or heterologous sequence having a GC content of approximately 40%.
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