(A) Amino acid sequence mapping analysis of parental capsid fragment crossovers in vectorized shuffled capsids. Library parents are depicted in different colors as indicated on the left. Large dots represent 100% parental match (i.e., the position in question matches only 1 parent), and small dots represent more than 1 parental match (i.e., the position matches more than 1 parent) at each position. The solid line for each chimera represents the library parents identified within the sequence between crossovers. A set of thin horizontal parallel lines between crossovers indicates that multiple parents match at an equal probability. A vertical spike indicates a single position switch between parents. VP1, VP2, VP3, and AAP open reading frames are shown below. (B) Amino acid sequence mapping analysis of parental AAP fragment crossovers in vectorized shuffled capsids. (C) The residues different from AAV3B of shuffled variants were 3-dimensionally false-color mapped onto the crystal structure of AAV6 VP3. Light gray residues correspond to AAV3B amino acids, while colored residues indicate surface-exposed amino acids derived from other serotypes. With the exception of AAV3B, color coding is as in A and B. (D) Enrichment scores were calculated for each amino acid position in the sequence of each chimera by comparison of sequences from parental serotypes based on maximum likelihood. Library parents are depicted in different colors as shown.