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We found that some early CTL responses could cross-recognize their variants with high magnitudes even before the variant sequences were abundant in the viral population.

Four double (D4Y+M7K, D4Y+S11K, P5Y+M7K, P5Y+S11K), two triple (D4S+P5Y+M7K, D4S+P5Y+S11K) and one quadruple (D4S+P5Y+M7K+S11K) variant sequences were characterized with SPR.

Such tags harboring homopolymer stretches frequently carry similar sequences with one or two base changes in the original SuperSAGE data, while such variant sequences were not observed in HT-SuperSAGE (Table S4).

Finally, we observed recognition of some variants despite the fact that at that time, the variant sequences were not yet observed and responses to the founder epitopes and other major variants were not detected (variant ET9 of Vpr EI9 at 1501 DPS and A2DY9 of Env AY9 at 190 DPS; Figures 7A and 8D).

We observed CTL responses with high magnitudes that reacted with epitope variants even before the variant sequences were detected (or became abundant) in the sampled viral population (e.g., variants of Vpr EI9, Env YW9, Pol NY9, Pol TI8 and Gag QW11; Figures 7, 8A and 8C).

Variant sequences were identified in the invertebrate MACITs (see below).

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The relative affinity of ERRα for the wild-type and variant sequences was assessed in competition binding studies using increasing amounts of unlabeled G or A dsDNA probe to displace wild-type G probe in the EMSA binding reactions.

The library of thioredoxin variant sequences was constructed by using gene assembly mutagenesis [ 22].

Variant sequences are identified and enumerated within the unselected and selected libraries.

Sequences shared between multiple isoforms are retained as unique contigs, and any adjacent variant sequences are split off as their own unique contigs.

Twenty different variant sequences are then created by introducing ten different sets of deletions and ten different sets of tandem duplications at known locations.

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