Because the main objective of this work was to test prove-of-concept computational strategies rather than to develop a practical application, a minimal ) . Their percentages were calculated by the formula, 100 * number of single mutations / 26 (data from Table 2 ). All the MPXV 26 isolates from the 2022 outbreak at Los Angeles USA coded for E353 . The A290V and A288P mutations were isolated in persistent MPXV infections despite Tecovirimat treatments 53, 54 . Single amino acid mutations were labeled to their left (wild-type amino acid+position number+mutation). Yellow vertical rectangle, mapped nearby-canonical phospholipase-like motif ( 312 NxKxxxxD) Green solid circles, mutations isolated in vitro by cell co-culture of several poxviruses with Tecovirimat . Red circles, single mutations identified from 26 patient isolates assayed for Tecovirimat-resistance. Red solid circles, mutations included into the F13L1 and F13L2 mutant models (Selected because they showed the highest i) percentages in the 26 patient isolates and ii) resistances > 500 µM Tecovirimat 30 ). number of two representative F13L mutant models containing the most abundant and higher Tecovirimat-resistant mutations were selected for these preliminary studies (Figure , mutation labelled red solid circles). Two models were minimally required because the N267D / N267deleted mutations exclude each other. None of them should be discarded due to their high frequencies and the high F13L structural impact the N267 deletion may have. In addition to the single mutations, the common 2022 background mutation E353K was also included. Therefore, the artificially designed pooled-mutant models were, F13L1 (coding for E353K + N267D + A288P + A290V + D294V + A295E + I372N) and F13L2 (coding for E353K + N267deleted + A288P + A290V + D294V + A295E + I372N).