Protocol

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protocol.pdf

Primer and gblock

Purpose Number Name Sequence Base Number
Construction of the A6-Ins15 and A6-Ins30 plasmids 1 NA-insert-F-59 CAGGACTCAACTTCAGTG 18
2 NA-insert-R-59 CCCAGCAACAACTTTATAG 19
3 PR8 N1-Ins15rec-F AAAGTTGTTGCTGGGAATCAGACATATGTTAACATCAGCAACACCAACTTTGCTGCTGGA 60
4 PR8 N1-Ins15rec-R TGAAGTTGAGTCCTGTCCAGCAGCAAAGTTGGTGTTGCTGATGTTAACATATGTCTGATT 60
5 PR8 N1-Ins30rec-F-58 AAAGTTGTTGCTGGGAATCAGACATATGTTAACATCAG 38
6 PR8 N1-Ins30rec-R-58 TGAAGTTGAGTCCTGCTTCTCTATGGTGGTGTTG 34
7 PR8 N1-Ins30-F AATCAGACATATGTTAACATCAGCAACACCAACTTTGCTGCTGGAAACACAACAGAGATAGTGTATCTGACCAACACCACCATAGAGAAG 90
8 PR8 N1-Ins30-R CTTCTCTATGGTGGTGTTGGTCAGATACACTATCTCTGTTGTGTTTCCAGCAGCAAAGTTGGTGTTGCTGATGTTAACATATGTCTGATT 90
Construction of A6-HAps and A4-NAps plasmids 9 HAps-5rec-F TCCGAAGTTGGGGGGAGCAAAAGCAGGGGAAAATAAAAACAACCAAATTGAAGGCATGAATCCAAACCAG 70
10 HAps-5rec-R CTGGTTTGGATTCATGCCTTCAATTTGGTTGTTTTTATTTTCCCCTGCTTTTGCTCCCCCCAACTTCGGA 70
11 HAps-3-F-63.6 TGGTGCTTTTGGTCTCCC 18
12 HAps-3-R-63.6 AGTAGAAACAAGGGTGTTTTTCC 23
13 HAps-3rec-F-63.6 ACCATTGACAAGTAGTGGTGCTTTTGGTCTCCC 33
14 HAps-3rec-R-63.6 GGCCGCCGGGTTATTAGTAGAAACAAGGGTGTTTTTCC 38
15 NAps-5-F-61.4 AGCGAAAGCAGGAGTTTAATTG 22
16 NAps-5-R-61.4 CCCAGCAACAACTTTATAGG 20
17 NAps-5rec-F-61.4 CCGAAGTTGGGGGGGAGCGAAAGCAGGAGTTTAATTG 37
18 NAps-5rec-R-61.4 TAGTTTTGCCTTCATCCCAGCAACAACTTTATAGG 35
19 NAps-3-F-65.9 GAGGCCTTGCTTCTGGGTTG 20
20 NAps-3-R-65.9 AGTAGAAACAAGGAGTTTTTTGAACAAACTAC 32
21 NAps-3rec-F-65.9 AGAATATGCATCTGAGAGGCCTTGCTTCTGGGTTG 35
22 NAps-3rec-R-65.9 GGGCCGCCGGGTTATTAGTAGAAACAAGGAGTTTTTTGAACAAACTAC 48
23 NA-5-line-F-67 ATGAATCCAAACCAGAAAATAATAACCATTGG 32
24 NA-5-line-R-67 CCCCCCAACTTCGGAGGT 18
25 NA-3-line-F-67 AATAACCCGGCGGCCCAA 18
26 NA-3-line-R-67 CTACTTGTCAATGGTGAACGGC 22
27 HA-5-line-F-68 ATGAAGGCAAAACTACTGGTCCTG 24
28 HA-5-line-R-68 CCCCCCCAACTTCGGAGG 18
29 HA-3-line-F-68 AATAACCCGGCGGCCCAA 18
30 HA-3-line-R-68 TCAGATGCATATTCTGCACTGCAAAG 26
Refinement of the A6-NAps plasmid 31 NA-5-PS-PM-F-60 ATGAATCCAAACCAGAAAATAATAAC 26
32 NA-5-PS-PM-R-60 TGCCTTCAATTTGGTTGTTTTTATTTTC 28
Construction of the A8-delNS1 plasmid 33 DelNS1-57-F GACATACTGATGAGGATG 18
34 DelNS1-57-R CTGAAAGCTTGACACAGTG 19
Construction of plasmids A6-SD9, A6-SD24, A6-SI14, and A6-SI28 35 WSN N1-Ins14rec-F ATTCAAACCGGAAATGACTCCCCCGCGAGCAACCAAGTAATGCCGTGTGAACCAATA 57
36 WSN N1-Ins14rec-R TCCAGTATGGTTTTGTATTGGTTCACACGGCATTACTTGGTTGCTCGCGGGGGAGTC 57
37 WSN N1-Ins28-F GACTCCCCCGCGAGCAACCAAGTAATGCCGTGTGAACCAATATCACAACCCGAAGCAACCAATGCAAGCCAAACAATAATAAAT 84
38 WSN N1-Ins28-R ATTTATTATTGTTTGGCTTGCATTGGTTGCTTCGGGTTGTGATATTGGTTCACACGGCATTACTTGGTTGCTCGCGGGGGAGTC 84
39 NA-insert2-F-56 CAAAACCATACTGGAATATG 20
40 NA-insert2-R-56 TTTCCGGTTTGAATTGAATG 20
41 WSN N1-Ins28rec-F-58 ATTCAAACCGGAAATGACTCCCCCGCGAGCAAC 33
42 WSN N1-Ins28rec-R-58 TCCAGTATGGTTTTGATTTATTATTGTTTGGCTTGCATTGGTTGCTTC 48
43 WSN N1-del9aa-F-59 GGCAGCATTACCTATAAAG 19
44 WSN N1-del9aa-R-59 ATTTCCGGTTTGAATTGAATG 21
45 WSN N1-del24aa-F-58 CAGGACTCAACTTCAGTG 18
46 WSN N1-del24aa-R-58 TGAATGGCTAATCCATATTG 20
47 WSN N1-Ins14rec2-F ATTCAAACCGGAAATGACTCCCCCGCGAGCAACCAAGTAATGCCGTGTGAACCAATACAAAACCATACTGGA 72
48 WSN N1-Ins14rec2-R TCCAGTATGGTTTTGTATTGGTTCACACGGCATTACTTGGTTGCTCGCGGGGGAGTCATTTCCGGTTTGAAT 72
Construction of A6-HAps(mut) and A4-NAps (mut) plasmids 49 HA-5-PS-PM-56-F GGATGAAGGCcAAACTACTGG 21
50 HA-5-PS-PM-56-R CAGCAACAACTTTATAGG 18
51 HA-3-PS-line-56-F GAGGCCTTGCTTCTGGGTTG 20
52 HA-3-PS-line-56-R TGAACTGGCGACAGTTGAG 19
53 NA-5-PS-line-60.4-F CAGGACTCAACTTCAGTG 18
54 NA-5-PS-line-60.4-R TGCCTTCAATTTGGTTGTTTTTATTTTC 28
55 NA-3-PS-line-64.6-F TGGTGCTTTTGGTCTCCC 18
52 NA-3-PS-line-64.6-R ACAGTCTAGCCCTGTTAGC 19
Construction of the A6-SI28-new plasmid 57 NA-28line-F-59.5 CAAAACCATACTGGAATATG 20
58 NA-28line-R-59.5 ATTGGTTCACACGGCATTAC 20
59 NA-28rec-F CCGTGTGAACCAATAAATGCAAGCCAAACAATAATAAATAACTATTATAATGACACACAAAACCATACTGGA 72
60 NA-28rec-R TCCAGTATGGTTTTGTGTGTCATTATAATAGTTATTTATTATTGTTTGGCTTGCATTTATTGGTTCACACGG 72
Primers for amplification of viral HA and NA protein genes 61 HA-cDNA-F-57.2 ATGAAGGCAAAACTACTG 18
62 HA-cDNA-R-57.2 TCAGATGCATATTCTGCAC 19
63 NA-vRNAps-F-61.8 AGCGAAAGCAGGAGTTTAATTG 22
64 NA-vRNA-F-61.8 AGCGAAAGCAGGAGTTTAAATG 22
65 NA-vRNA-R-61.8 AGTAGAAACAAGGAGTTTTTTGAAC 25
66 NA-cDNA-F-58.3 ATGAATCCAAACCAGAAAATAATAAC 26
67 NA-cDNA-R-58.3 CTACTTGTCAATGGTGAAC 19
68 HA-vRNA-F-61.1 AGCAAAAGCAGGGGAAAATAAAAAC 25
69 HA-vRNA-R-61.1 AGTAGAAACAAGGGTGTTTTTC 22
Primers for sequencing 70 NA-IS300-R TGCACTTGCTGACCAAGC 18
71 HA-IS245-R AGCAGTGAGTCGCATTCT 18
72 NA-RS200-F ATCGGTCAGGGTACAGCG 18
73 HA-RS200-F GTGTAAGAAATGGGACTT 18
74 WSN-F AGCTAGCGGCCGATCCCC 18
75 WSN-R CAGAGGGCCCCGGGCGCT 18
76 A8del-F ACACTTTCGGACATCTGG 18
77 A8del-R GGACAGGTGTCCGTGTCG 18
Primers for qPCR 78 M1-qPCR-F GACCAATCCTGTCACCTC 18
79 M1-qPCR-R GATCTCCGTTCCCATTAAGAG 21
80 GAPDH-qPCR-F GAAGGTGAAGGTCGGAGTC 19
81 GAPDH-qPCR-R GAAGATGGTGATGGGATTTC 20
gblock Number Name Sequence Length
NA vRNA 1 NA segment AGCGAAAGCAGGAGTTTAAATGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATCAAAACCATACTGGAATATGCAACCAAGGCAGCATTACCTATAAAGTTGTTGCTGGGCAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 1409
HA vRNA 2 HA segment AGCAAAAGCAGGGGAAAATAAAAACAACCAAAATGAAGGCAAAACTACTGGTCCTGTTATATGCATTTGTAGCTACAGATGCAGACACAATATGTATAGGCTACCATGCGAACAACTCAACCGACACTGTTGACACAATACTCGAGAAGAATGTGGCAGTGACACATTCTGTTAACCTGCTCGAAGACAGCCACAACGGGAAACTATGTAAATTAAAAGGAATAGCCCCACTACAATTGGGGAAATGTAACATCACCGGATGGCTCTTGGGAAATCCAGAATGCGACTCACTGCTTCCAGCGAGATCATGGTCCTACATTGTAGAAACACCAAACTCTGAGAATGGAGCATGTTATCCAGGAGATCTCATCGACTATGAGGAACTGAGGGAGCAATTGAGCTCAGTATCATCATTAGAAAGATTCGAAATATTTCCCAAGGAAAGTTCATGGCCCAACCACACATTCAACGGAGTAACAGTATCATGCTCCCATAGGGGAAAAAGCAGTTTTTACAGAAATTTGCTATGGCTGACGAAGAAGGGGGATTCATACCCAAAGCTGACCAATTCCTATGTGAACAATAAAGGGAAAGAAGTCCTTGTACTATGGGGTGTTCATCACCCGTCTAGCAGTGATGAGCAACAGAGTCTCTATAGTAATGGAAATGCTTATGTCTCTGTAGCGTCTTCAAATTATAACAGGAGATTCACCCCGGAAATAGCTGCAAGGCCCAAAGTAAGAGATCAACATGGGAGGATGAACTATTACTGGACCTTGCTAGAACCCGGAGACACAATAATATTTGAGGCAACTGGTAATCTAATAGCACCATGGTATGCTTTCGCACTGAGTAGAGGGTTTGAGTCCGGCATCATCACCTCAAACGCGTCAATGCATGAGTGTAACACGAAGTGTCAAACACCCCAGGGAGCTATAAACAGCAATCTCCCTTTCCAGAATATACACCCAGTCACAATAGGAGAGTGCCCAAAATATGTCAGGAGTACCAAATTGAGGATGGTTACAGGACTAAGAAACATCCCATCCATTCAATACAGAGGTCTATTTGGAGCCATTGCTGGTTTTATTGAGGGGGGATGGACTGGAATGATAGATGGATGGTATGGTTATCATCATCAGAATGAACAGGGATCAGGCTATGCAGCGGATCAAAAAAGCACACAAAATGCCATTAACGGGATTACAAACAAGGTGAACTCTGTTATCGAGAAAATGAACACTCAATTCACAGCTGTGGGTAAAGAATTCAACAACTTAGAAAAAAGGATGGAAAATTTAAATAAAAAAGTTGATGATGGGTTTCTGGACATTTGGACATATAATGCAGAATTGTTAGTTCTACTGGAAAATGAAAGGACTTTGGATTTCCATGACTTAAATGTGAAGAATCTGTACGAGAAAGTAAAAAGCCAATTAAAGAATAATGCCAAAGAAATCGGAAATGGGTGTTTTGAGTTCTACCACAAGTGTGACAATGAATGCATGGAAAGTGTAAGAAATGGGACTTATGATTATCCAAAATATTCAGAAGAATCAAAGTTGAACAGGGAAAAGATAGATGGAGTGAAATTGGAATCAATGGGGGTGTATCAGATTCTGGCGATCTACTCAACTGTCGCCAGTTCACTGGTGCTTTTGGTCTCCCTGGGGGCAATCAGTTTCTGGATGTGTTCTAATGGGTCTTTGCAGTGCAGAATATGCATCTGAGATTAGGATTTCAGAAATATAAGGAAAAACACCCTTGTTTCTACT 1775
NS vRNA 3 NS segment AGCAAAAGCAGGGTGACAAAGACATAATGGATCCAAACACTGTGTCAAGCTTTCAGGTAGATTGCTTTCTTTGGCATGTCCGCAAAAGAGTTGCAGACCAAGAACTAGGTGATGCCCCATTCCTTGATCGGCTTCGCCGAGATCAGAAGTCCCTAAGAGGAAGAGGCAGCACTCTTGGTCTGGACATCGAAACAGCCACCCGTGCTGGAAAGCAAATAGTGGAGCGGATTCTGAAGGAAGAATCTGATGAGGCACTCAAAATGACCATGGCCTCTGTACCTGCATCGCGCTACCTAACTGACATGACTCTTGAGGAAATGTCAAGGCACTGGTTCATGCTCATGCCCAAGCAGAAAGTGGCAGGCCCTCTTTGTATCAGAATGGACCAGGCGATCATGGATAAGAACATCATACTGAAAGCGAACTTCAGTGTGATTTTTGACCGGCTGGAGACTCTAATATTACTAAGGGCCTTCACCGAAGAGGGGACAATTGTTGGCGAAATTTCACCACTGCCCTCTCTTCCAGGACATACTGATGAGGATGTCAAAAATGCAGTTGGGGTCCTCATCGGAGGACTTGAATGGAATAATAACACAGTTCGAGTCTCTGAAACTCTACAGAGATTCGCTTGGAGAAGCAGTAATGAGAATGGGAGACCTCCACTCACTCCAAAACAGAAACGGAAAATGGCGGGAACAATTAGGTCAGAAGTTTGAAGAAATAAGATGGTTGATTGAAGAAGTGAGACACAGACTGAAGATAACAGAGAATAGTTTTGAGCAAATAACATTTATGCAAGCCTTACAACTATTGCTTGAAGTGGAGCAAGAGATAAGAACTTTCTCGTTTCAGCTTATTTAATAATAAAAAACACCCTTGTTTCTACT 890
NA vRNA after the exchange of packaging signals 4 NA-HAps AGCAAAAGCAGGGGAAAATAAAAACAACCAAATTGAAGGCAATGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATCAAAACCATACTGGAATATGCAACCAAGGCAGCATTACCTATAAAGTTGTTGCTGGGCAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTGGTGCTTTTGGTCTCCCTGGGGGCAATCAGTTTCTGGATGTGTTCTAATGGGTCTTTGCAGTGCAGAATATGCATCTGAGATTAGGATTTCAGAAATATAAGGAAAAACACCCTTGTTTCTACT 1528
HA vRNA after the exchange of packaging signals 5 HA-HAps AGCGAAAGCAGGAGTTTAATTGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATCAAAACCATACTGGAATATGCAACCAAGGCAGCATTACCTATAAAGTTGTTGCTGGGATGAAGGCAAAACTACTGGTCCTGTTATATGCATTTGTAGCTACAGATGCAGACACAATATGTATAGGCTACCATGCGAACAACTCAACCGACACTGTTGACACAATACTCGAGAAGAATGTGGCAGTGACACATTCTGTTAACCTGCTCGAAGACAGCCACAACGGGAAACTATGTAAATTAAAAGGAATAGCCCCACTACAATTGGGGAAATGTAACATCACCGGATGGCTCTTGGGAAATCCAGAATGCGACTCACTGCTTCCAGCGAGATCATGGTCCTACATTGTAGAAACACCAAACTCTGAGAATGGAGCATGTTATCCAGGAGATCTCATCGACTATGAGGAACTGAGGGAGCAATTGAGCTCAGTATCATCATTAGAAAGATTCGAAATATTTCCCAAGGAAAGTTCATGGCCCAACCACACATTCAACGGAGTAACAGTATCATGCTCCCATAGGGGAAAAAGCAGTTTTTACAGAAATTTGCTATGGCTGACGAAGAAGGGGGATTCATACCCAAAGCTGACCAATTCCTATGTGAACAATAAAGGGAAAGAAGTCCTTGTACTATGGGGTGTTCATCACCCGTCTAGCAGTGATGAGCAACAGAGTCTCTATAGTAATGGAAATGCTTATGTCTCTGTAGCGTCTTCAAATTATAACAGGAGATTCACCCCGGAAATAGCTGCAAGGCCCAAAGTAAGAGATCAACATGGGAGGATGAACTATTACTGGACCTTGCTAGAACCCGGAGACACAATAATATTTGAGGCAACTGGTAATCTAATAGCACCATGGTATGCTTTCGCACTGAGTAGAGGGTTTGAGTCCGGCATCATCACCTCAAACGCGTCAATGCATGAGTGTAACACGAAGTGTCAAACACCCCAGGGAGCTATAAACAGCAATCTCCCTTTCCAGAATATACACCCAGTCACAATAGGAGAGTGCCCAAAATATGTCAGGAGTACCAAATTGAGGATGGTTACAGGACTAAGAAACATCCCATCCATTCAATACAGAGGTCTATTTGGAGCCATTGCTGGTTTTATTGAGGGGGGATGGACTGGAATGATAGATGGATGGTATGGTTATCATCATCAGAATGAACAGGGATCAGGCTATGCAGCGGATCAAAAAAGCACACAAAATGCCATTAACGGGATTACAAACAAGGTGAACTCTGTTATCGAGAAAATGAACACTCAATTCACAGCTGTGGGTAAAGAATTCAACAACTTAGAAAAAAGGATGGAAAATTTAAATAAAAAAGTTGATGATGGGTTTCTGGACATTTGGACATATAATGCAGAATTGTTAGTTCTACTGGAAAATGAAAGGACTTTGGATTTCCATGACTTAAATGTGAAGAATCTGTACGAGAAAGTAAAAAGCCAATTAAAGAATAATGCCAAAGAAATCGGAAATGGGTGTTTTGAGTTCTACCACAAGTGTGACAATGAATGCATGGAAAGTGTAAGAAATGGGACTTATGATTATCCAAAATATTCAGAAGAATCAAAGTTGAACAGGGAAAAGATAGATGGAGTGAAATTGGAATCAATGGGGGTGTATCAGATTCTGGCGATCTACTCAACTGTCGCCAGTTCACTGGTGCTTTTGGTCTCCCTGGGGGCAATCAGTTTCTGGATGTGTTCTAATGGGTCTTTGCAGTGCAGAATATGCATCTGAGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 2085
NA vRNA after exchanging packaging signals and making synonymous mutations 6 NA-HAps(mut) AGCAAAAGCAGGGGAAAATAAAAACAACCAAATTGAAGGCAATGAACCCCAACCAGAAAATCATCACCATTGGGAGCATCTGCATGGTGGTGGGCATTATTAGCCTGATTCTGCAGATCGGCAATATTATCAGCATCTGGATCTCACACAGCATCCAGACCGGAAACCAGAACCACACCGGCATCTGCAACCAGGGCAGCATCACCTACAAGGTGGTGGCCGGCCAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCATGCTTTTGGGTGGAGCTGATCAGGGGGCTGCCCGAGGAGGATGCCATCTGGACCAGTGGAAGCATTATCAGCTTTTGCGGCGTGAACAGCGACACCGTGGACTGGTCCTGGCCAGATGGCGCCGAGCTGCCTTTCACCATCGACAAGTGATGGTGCTTTTGGTCTCCCTGGGGGCAATCAGTTTCTGGATGTGTTCTAATGGGTCTTTGCAGTGCAGAATATGCATCTGAGATTAGGATTTCAGAAATATAAGGAAAAACACCCTTGTTTCTACT 1528
HA vRNA after exchanging packaging signals and making synonymous mutations 7 HA-HAps(mut) AGCGAAAGCAGGAGTTTAATTGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATCAAAACCATACTGGAATATGCAACCAAGGCAGCATTACCTATAAAGTTGTTGCTGGGATGAAGGCCAAACTACTGGTCCTGTTATATGCATTTGTAGCTACAGATGCAGACACAATATGTATAGGCTACCATGCGAACAACTCAACCGACACTGTTGACACAATACTCGAGAAGAATGTGGCAGTGACACATTCTGTTAACCTGCTCGAAGACAGCCACAACGGGAAACTATGTAAATTAAAAGGAATAGCCCCACTACAATTGGGGAAATGTAACATCACCGGATGGCTCTTGGGAAATCCAGAATGCGACTCACTGCTTCCAGCGAGATCATGGTCCTACATTGTAGAAACACCAAACTCTGAGAATGGAGCATGTTATCCAGGAGATCTCATCGACTATGAGGAACTGAGGGAGCAATTGAGCTCAGTATCATCATTAGAAAGATTCGAAATATTTCCCAAGGAAAGTTCATGGCCCAACCACACATTCAACGGAGTAACAGTATCATGCTCCCATAGGGGAAAAAGCAGTTTTTACAGAAATTTGCTATGGCTGACGAAGAAGGGGGATTCATACCCAAAGCTGACCAATTCCTATGTGAACAATAAAGGGAAAGAAGTCCTTGTACTATGGGGTGTTCATCACCCGTCTAGCAGTGATGAGCAACAGAGTCTCTATAGTAATGGAAATGCTTATGTCTCTGTAGCGTCTTCAAATTATAACAGGAGATTCACCCCGGAAATAGCTGCAAGGCCCAAAGTAAGAGATCAACATGGGAGGATGAACTATTACTGGACCTTGCTAGAACCCGGAGACACAATAATATTTGAGGCAACTGGTAATCTAATAGCACCATGGTATGCTTTCGCACTGAGTAGAGGGTTTGAGTCCGGCATCATCACCTCAAACGCGTCAATGCATGAGTGTAACACGAAGTGTCAAACACCCCAGGGAGCTATAAACAGCAATCTCCCTTTCCAGAATATACACCCAGTCACAATAGGAGAGTGCCCAAAATATGTCAGGAGTACCAAATTGAGGATGGTTACAGGACTAAGAAACATCCCATCCATTCAATACAGAGGTCTATTTGGAGCCATTGCTGGTTTTATTGAGGGGGGATGGACTGGAATGATAGATGGATGGTATGGTTATCATCATCAGAATGAACAGGGATCAGGCTATGCAGCGGATCAAAAAAGCACACAAAATGCCATTAACGGGATTACAAACAAGGTGAACTCTGTTATCGAGAAAATGAACACTCAATTCACAGCTGTGGGTAAAGAATTCAACAACTTAGAAAAAAGGATGGAAAATTTAAATAAAAAAGTTGATGATGGGTTTCTGGACATTTGGACATATAATGCAGAATTGTTAGTTCTACTGGAAAATGAAAGGACTTTGGATTTCCATGACTTAAATGTGAAGAATCTGTACGAGAAAGTAAAAAGCCAATTAAAGAATAATGCCAAAGAAATCGGAAATGGGTGTTTTGAGTTCTACCACAAGTGTGACAATGAATGCATGGAAAGTGTAAGAAATGGGACTTATGATTATCCAAAATATTCAGAAGAATCAAAGTTGAACAGGGAAAAGATAGATGGAGTGAAATTGGAATCAATGGGGGTGTATCAGATTCTGGCGATCTACTCAACTGTCGCCAGTTCACTGGTGCTGCTGGTGAGCCTGGGCGCCATCTCTTTCTGGATGTGCAGCAACGGCTCCCTGCAGTGCAGAATCTGCATCTGAGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 2085
NS vRNA after deletion of NS1 8 NS delNS1 AGCAAAAGCAGGGTGACAAAGACATAATGGATCCAAACACTGTGTCAAGCTTTCAGGACATACTGATGAGGATGTCAAAAATGCAGTTGGGGTCCTCATCGGAGGACTTGAATGGAATAATAACACAGTTCGAGTCTCTGAAACTCTACAGAGATTCGCTTGGAGAAGCAGTAATGAGAATGGGAGACCTCCACTCACTCCAAAACAGAAACGGAAAATGGCGGGAACAATTAGGTCAGAAGTTTGAAGAAATAAGATGGTTGATTGAAGAAGTGAGACACAGACTGAAGATAACAGAGAATAGTTTTGAGCAAATAACATTTATGCAAGCCTTACAACTATTGCTTGAAGTGGAGCAAGAGATAAGAACTTTCTCGTTTCAGCTTATTTAATAATAAAAAACACCCTTGTTTCTACT 418
NA vRNA after insertion of 15 amino acids 9 NA-Ins15 AGCGAAAGCAGGAGTTTAAATGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATCAAAACCATACTGGAATATGCAACCAAGGCAGCATTACCTATAAAGTTGTTGCTGGGAATCAGACATATGTTAACATCAGCAACACCAACTTTGCTGCTGGACAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 1454
NA vRNA after insertion of 30 amino acids 10 NA-Ins30 AGCGAAAGCAGGAGTTTAAATGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATCAAAACCATACTGGAATATGCAACCAAGGCAGCATTACCTATAAAGTTGTTGCTGGGAATCAGACATATGTTAACATCAGCAACACCAACTTTGCTGCTGGAAACACAACAGAGATAGTGTATCTGACCAACACCACCATAGAGAAGCAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 1499
NA vRNA after deletion of 24 amino acids 11 NA-SD24 AGCGAAAGCAGGAGTTTAAATGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCACAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 1337
NA vRNA after deletion of 9 amino acids 12 NA-SD9 AGCGAAAGCAGGAGTTTAAATGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATGGCAGCATTACCTATAAAGTTGTTGCTGGGCAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 1382
NA vRNA after insertion of 14 amino acids 13 NA-SI14 AGCGAAAGCAGGAGTTTAAATGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATGACTCCCCCGCGAGCAACCAAGTAATGCCGTGTGAACCAATACAAAACCATACTGGAATATGCAACCAAGGCAGCATTACCTATAAAGTTGTTGCTGGGCAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 1451
NA vRNA after insertion of 28 amino acids 14 NA-SI28 AGCGAAAGCAGGAGTTTAAATGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATGACTCCCCCGCGAGCAACCAAGTAATGCCGTGTGAACCAATATCACAACCCGAAGCAACCAATGCAAGCCAAACAATAATAAATCAAAACCATACTGGAATATGCAACCAAGGCAGCATTACCTATAAAGTTGTTGCTGGGCAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 1493
NA vRNA after insertion of new 28 amino acids 15 NA-SI28-new AGCGAAAGCAGGAGTTTAAATGAATCCAAACCAGAAAATAATAACCATTGGGTCAATCTGTATGGTAGTCGGAATAATTAGCCTAATATTGCAAATAGGAAATATAATCTCAATATGGATTAGCCATTCAATTCAAACCGGAAATGACTCCCCCGCGAGCAACCAAGTAATGCCGTGTGAACCAATAAATGCAAGCCAAACAATAATAAATAACTATTATAATGACACACAAAACCATACTGGAATATGCAACCAAGGCAGCATTACCTATAAAGTTGTTGCTGGGCAGGACTCAACTTCAGTGATATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATACACAGCAAAGACAATGGCATAAGAATTGGTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCTTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTTCTGACTCAAGGCGCCTTACTGAATGACAAGCATTCAAGGGGGACCTTTAAGGACAGAAGCCCTTATAGGGCCTTAATGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGGTTTGAATCGGTTGCTTGGTCAGCAAGTGCATGTCATGATGGAATGGGCTGGCTAACAATCGGAATTTCTGGTCCAGATGATGGAGCAGTGGCTGTATTAAAATACAACCGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAATATATTGAGAACACAAGAGTCTGAATGTACCTGTGTAAATGGTTCATGTTTTACCATAATGACCGATGGCCCAAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATCGAGAAGGGGAAGGTTACTAAATCGATAGAGTTGAATGCACCTAATTCTCACTACGAGGAATGTTCCTGTTACCCTGATACCGGCAAAGTGATGTGTGTGTGCAGAGACAATTGGCACGGTTCGAACCGACCATGGGTGTCCTTCGACCAAAACCTAGATTATAAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCAAAGATGGAACAGGCAGCTGTGGCCCAGTGTCTGCTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAAGTATGGCAATGGTGTTTGGATAGGAAGGACTAAAAGTGACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACTGATAGTAGGTTCTCTATGAGACAAGATGTTGTGGCAATAACTAATCGGTCAGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGCTAACAGGGCTAGACTGTATGAGGCCTTGCTTCTGGGTTGAATTAATCAGGGGGCTACCTGAGGAGGACGCAATCTGGACTAGTGGGAGCATCATTTCTTTTTGTGGTGTGAATAGTGATACTGTAGATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAGTTTGTTCAAAAAACTCCTTGTTTCTACT 1493