(Sample_Name = F394Lck-2_T7_7) ACCCCAACGAGGCTGCGTTTAACGGGCCCTCTAGACTCGAGACCATG GGCTGTGGCTGCAGCTCACACCCGGAA GATGAC TGGATGGAAAACATCGATGTGTGTGAGAACTGCCATTATCCCATAGTC CCACTGGATGGCAAGGGCACGCTGCTCA TCCG AAATGGCTCTGAGGTGCGGGACCCACTGGTTACCTACGAAGGCTCCA ATCCGCCGGCTTCCCCACTGCAAGACAAC CTGG TTATCGCTCTGCACAGCTATGAGCCCTCTCACGACGGAGATCTGGGC TTTGAGAAGGGGGAACAGCTCCGCATCCT GGAG CAGAGCGGCGAGTGGTGGAAGGCGCAGTCCCTGACCACGGGCCAGG

AAGGCTTCATCCCCTTCAATTTTGTGGCCA AAGC GAACAGCCTGGAGCCCGAACCCTGGTTCTTCAAGAACCTGAGCCGCA AGGACGCGGAGCGGCAGCTCCTGGCGCCC GGGA ACACTCACGGCTCCTTCCTCATCCGGGAGAGCGAGAGCACCGCGGGA TCGTTTTCACTGTCGGTCCGGGACTTCGA CCAG AACCAGGGAGAGGTGGTGAAACATTACAAGATCCGTAATCTGGACAA CGGTGGCTTCTACATCTCCCCTCGAATCA CTTT TCCCGGCCTGCATGAACTGGTCCGCCATTACACCAATGCTTCAGATG GGCTGTGCACACGGTTGAGCCGCCCCTGC CAGA CCCAGAAGCCCCAGAAGCCGTGGTGGGAGGACGAGTGGGAGGTTCC CAGGGAGACGCTGAAGCTGGTGGAGCGGCT GGGG GCTGGACAGTTCGGGGAGGTGTGGATGGGGTACTACAACGGGCACA CGAAGGTGGCGGTGAAGAGCCTGAAGCAGG CAGC ATGTCCCCGAACGCCTTCCTGGCCGAGGCCAACCTCATGAAGCAGCT GCAACACCAGCGGCTGGTTCGGCTCTACG CTGT GGTCACCCAGGAGCCCATCTACATCATCACTGAATACATGGAGAATG GGAGTCTAGTGGATTTTCTCAGACCCTTC AGCA TCAGTGACATCAACAACTCCTGGACATGCAGCCAAATGCAGAGCATG

CATCATGAGAGCGATTATATCATCGTGAC TCGG CTGCAACATCTGGTGTCTGACACCTGACTGCAAGATGCGACTTGCTAG CCGCTCATGAAGACAACGAGTACCAGCA GGAG GCAGTTCATAAGGTGAAGCGAGCATACTCGGAATACATAGTCAATTG CTGGAATCTCGTGACGTATGTACAGGCAT CTTA CAGGGCATCACACCGGGAGAATATAATCATCGGCGGGAAGGG

(Sample_Name = F394Lck-2_BGH_8) TGGCACTGCTACTTAAGCTGGAGACGTTTAACTCAATGATGATGATG ATGATGGTCGACGGCGCTATTCAGATC CTCTTC TGAGATGAGTTTTTGTTCTAGAAAGCTTGGCTGAGGCTGGTACTGGCC CTCTGTGGCCGTGAAGAAGTCCTCCAGC ACAC TGCGCAGGTAGTCAAAGGTGGGCCGGTCCTCTGGGCGCTCCTTCCAG CACAGCCTCATGAGTTGGTACAGCTCCTC TGGA CAGTTGTCAGGGCGCACCATGCGGTAGCCTCGCTCCAGGTTCTGAAT CACCTCCGGGTTGGTCATCCCTGGGTAAG GGAT GCGGCCGTGGGTGACAATTTCCGTCAGCAGGATCCCAAAAGACCACA CATCTGACTTGATGGTGAATGTCCCGTAG TTAA TGGCTTCTGGCGCTGTCCACTTAATGGGAAACTTGGCCCCCTCCCTG GCTGTGTACTCGTTGTCCTCAATGAGGCG TGCT AGGCCAAAGTCTGCAATCTTGCAGCTCAGGGTGTCAGACACCAGAAT GTTGGCAGCCCGAAGGTCACGATGAATAT AATT CCGCTCTTCAATGAATGCCATGCCTTCTGCAATTTGGGCTGCCATGTC CAGGAGTTTGTTGATGGTCAACTTGATG CCTG AAGGGGTCTTGAGAAAATCCACTAGACTCCCATTCTCCATGTATTCAG TGATGATGTAGATGGGCTCCTGGGTGAC

CACA GCGTAGAGCCGAACCAGCCGCTGGTGTTGCAGCTGCTTCATGAGGTT GGCCTCGGCCAGGAAGGCGTTCGGGGACA TGCT GCCCTGCTTCAGGCTCTTCACCGCCACCTTCGTGTGCCCGTTGTAGTA CCCCATCCACACCTCCCCGAACTGTCCA GCCC CCAGCCGCTCCACCAGCTTCAGCGTCTCCCTGGGAACCTCCCACTCG TCCTCCCACCACGGCTTCTGGGGCTTCTG GGTC TGGCAGGGGCGGCTCAACCGTGTGCACAGCCCATCTGAAGCATTGGT GTATGGCGGACCAGTTCATGCAGCCGGGA AAGT GATCGAGGGGAGATGTAGAAGCCACGTGTCAGATACGATCTGTATGT TCACCACCTCTCCTGTCTGTCGAGTCCGA CGAC GTGAAACGATCCGCGGTGCTCCCGCTTTCCGATGAGAAGACTGAGTG TTCCGGGCCAGGACTGCGCTCCGTCTGCG CTCA GTCTGAGAAAAGGTCCGTCAGCGTTCTTGCCCAATGAGGGTAAGCCT GCCGTCAGAGTGCCTCAACTCGGCTGTCC GAGA TGAGCAGTTCTCCTTTAAGACAGAATATCTCTT

> (5' --> 3' complementary sequence) - 1313 nucleotides aagagatattctgtcttaaaggagaactgctcatctcggacagccgagtt gaggcactctgacggcaggcttaccctcattgggcaagaacgctgacgga ccttttctcagactgagcgcagacggagcgcagtcctggcccggaacact cagtcttctcatcggaaagcgggagcaccgcggatcgtttcacgtcgtcg gactcgacagacaggagaggtggtgaacatacagatcgtatctgacacgt ggcttctacatctcccctcgatcactttcccggctgcatgaactggtccg ccatacaccaatgcttcagatgggctgtgcacacggttgagccgcccctg ccagacccagaagccccagaagccgtggtgggaggacgagtgggaggttc ccagggagacgctgaagctggtggagcggctgggggctggacagttcggg gaggtgtggatggggtactacaacgggcacacgaaggtggcggtgaagag cctgaagcagggcagcatgtccccgaacgccttcctggccgaggccaacc tcatgaagcagctgcaacaccagcggctggttcggctctacgctgtggtc acccaggagcccatctacatcatcactgaatacatggagaatgggagtct agtggattttctcaagaccccttcaggcatcaagttgaccatcaacaaac tcctggacatggcagcccaaattgcagaaggcatggcattcattgaagag cggaattatattcatcgtgaccttcgggctgccaacattctggtgtctga

caccctgagctgcaagattgcagactttggcctagcacgcctcattgagg acaacgagtacacagccagggagggggccaagtttcccattaagtggaca gcgccagaagccattaactacgggacattcaccatcaagtcagatgtgtg gtcttttgggatcctgctgacggaaattgtcacccacggccgcatccctt acccagggatgaccaacccggaggtgattcagaacctggagcgaggctac cgcatggtgcgccctgacaactgtccagaggagctgtaccaactcatgag gctgtgctggaaggagcgcccagaggaccggcccacctttgactacctgc gcagtgtgctggaggacttcttcacggccacagagggccagtaccagcct cagccaagctttctagaacaaaaactcatctcagaagaggatctgaatag cgccgtcgaccatcatcatcatcatcattgagttaaacgtctccagctta agtagcagtgcca Conclusion: hLck, T7 shows G278A and S279A, BGH shows L216M, D283N, no mutation at 394. (Sample_Name = F394Lck-4_T7_9) AAAAAACCGTGGCTGCGTTTACGGGCCCTCTAGACTCGAGACCATGG GCTGTGGCTGCAGCTCACACCCGGAAG ATGACT GGATGGAAAACATCGATGTGTGTGAGAACTGCCATTATCCCATAGTCC CACTGGATGGCAAGGGCACGCTGCTCAT CCGA AATGGCTCTGAGGTGCGGGACCCACTGGTTACCTACGAAGGCTCCAA TCCGCCGGCTTCCCCACTGCAAGACAACC TGGT TATCGCTCTGCACAGCTATGAGCCCTCTCACGACGGAGATCTGGGCT TTGAGAAGGGGGAACAGCTCCGCATCCTG GAGC AGAGCGGCGAGTGGTGGAAGGCGCAGTCCCTGACCACGGGCCAGGA AGGCTTCATCCCCTTCAATTTTGTGGCCAA AGCG AACAGCCTGGAGCCCGAACCCTGGTTCTTCAAGAACCTGAGCCGCAA GGACGCGGAGCGGCAGCTCCTGGCGCCCG GGAA CACTCACGGCTCCTTCCTCATCCGGGAGAGCGAGAGCACCGCGGGAT CGTTTTCACTGTCGGTCCGGGACTTCGAC CAGA ACCAGGGAGAGGTGGTGAAACATTACAAGATCCGTAATCTGGACAAC GGTGGCTTCTACATCTCCCCTCGAATCAC TTTT

CCCGGCCTGCATGAACTGGTCCGCCATTACACCAATGCTTCAGATGG GCTGTGCACACGGTTGAGCCGCCCCTGCC AGAC CCAGAAGCCCCAGAAGCCGTGGTGGGAGGACGAGTGGGAGGTTCCC AGGGAGACGCTGAAGCTGGTGGAGCGGCTG GGGG CTGGACAGTTCGGGGAGGTGCGGATGGGGTACTACAACGGGCACAC GAAGGTGGCGGTGAAGAGCCTGAAGCAGGG CAGC ATGTCCCCGAACGCCTTCCTGGCCGAGGCCAACCTCATGAAGCAGCT GCAACACCAGCGGCTGGTTCGGCTCTACG CTGT GGTCACCCAGGAGCCCATCTACATCATCACTGATACATGAGAATGGG AGTCTAGTGATTTTCTCAGACCCTTCAGC ATCA GTGACATCACAACTCTGACATGCAGCCCAAATGCAGAGCATGCATCAT GAAGAGCGATATTTCATCGTGACTCGGC TGCA ACATCTGTGTCTGACCCTGACTGCAGATGCGACTTGCTACACGCCTCA TGAGACACGAGATACAGCAGGAGGGTAG TTCA ATATGAAGGCGAGCATACTCGATCACTAGTCAAGTGCTTGAATCTGTC TCGATTCCAGGCGATCTTTACAGGATAT CACC CGGAGGTATATCTACTACGTA

(Sample_Name = F394Lck-4_BGH_10) CGGGCATATTACTTAGCTGGAGACGTTTAACTCAATGATGATGATGA TGATGGTCGACGGCGCTATTCAGATCC TCTTCT GAGATGAGTTTTTGTTCTAGAAAGCTTGGCTGAGGCTGGTACTGGCC CTCTGTGGCCGTGAAGAAGTCCTCCAGCA CACT GCGCAGGTAGTCAAAGGTGGGCCGGTCCTCTGGGCGCTCCTTCCAG CACAGCCTCATGAGTTGGTACAGCTCCTCT GGAC AGTTGTCAGGGCGCACCATGCGGTAGCCTCGCTCCAGGTTCTGAATC

ACCTCCGGGTTGGTCATCCCTGGGTAAGG GATG CGGCCGTGGGTGACAATTTCCGTCAGCAGGATCCCAAAAGACCACAC ATCTGACTTGATGGTGAATGTCCCGTAGT TAAT GGCTTCTGGCGCTGTCCACTTAATGGGAAACTTGGCCCCCTCCCTGG CTGTGTACTCGTTGTCCTCAATGAGGCGT GCTA GGCCAAAGTCTGCAATCTTGCAGCTCAGGGTGTCAGACACCAGAATG TTGGCAGCCCGAAGGTCACGATGAAAATA ATTC CGCTCTTCAATGAATGCCATGCCTTCTGCAATTTGGGCTGCCATGTCC AGGAGTTTGTTGATGGTCAACTTGATGC CTGA AGGGGTCTTGAGAAAATCCACTAGACTCCCATTCTCCATGTATTCAGT GATGATGTAGATGGGCTCCTGGGTGACC ACAG CGTAGAGCCGAACCAGCCGCTGGTGTTGCAGCTGCTTCATGAGGTTG GCCTCGGCCAGGAAGGCGTTCGGGGACAT GCTG CCCTGCTTCAGGCTCTTCACCGCCACCTTCGTGTGCCCGTTGTAGTAC CCCATCCGCACCTCCCCGAACTGTCCAG CCCC CAGCCGCTCCACCAGCTTCAGCGTCTCCCTGGGAACCTCCCACTCGT CCTCCCACCACGGCTTCTGGGGCTTCTGG GTCT GGCAGGGCGGCTCAACCGTGTGCACAGCCCATCTGAAGCATTGGTGT AATGCGGACCAGTTCATGCAGCCGGGAAA GTGA TTCGAGGGGAGAATGTAGAAGCCACCGTTGTCAAGATACGATCTGTA ATGTTCACCACCTCCCCCTGTTCTGTCGA GTCC CGAACGACGTGAAAACGATCCCGCGGTGCTCTCGCTCTCCGAAAAAG AAAGAACTGAGTTTCCCGGGCCAGACTGC CGCT CCGTCTGCGCCTCAGTTCTTGAAAACAGGTTCGTCCAAGCGTTCCCTG CCAAAAATGAAGTAAGCTCTGCGGTCAG GAGA GTGCCTCACATCGCGCTGTTCAAGAAGAGAGAAGATTTCCTCTCTCTT CAACAGAAAT > (5' --> 3' complementary sequence) 1338

nucleotides atttctgttgaagagagaggaaatcttctctcttcttgaacagcgcgatg tgaggcactctcctgaccgcagagcttacttcatttttggcagggaacgc ttggacgaacctgttttcaagaactgaggcgcagacggagcggcagtctg gcccgggaaactcagttctttctttttcggagagcgagagcaccgcggga tcgttttcacgtcgttcgggactcgacagaacagggggaggtggtgaaca ttacagatcgtatcttgacaacggtggcttctacattctcccctcgaatc actttcccggctgcatgaactggtccgcattacaccaatgcttcagatgg gctgtgcacacggttgagccgccctgccagacccagaagccccagaagcc gtggtgggaggacgagtgggaggttcccagggagacgctgaagctggtgg agcggctgggggctggacagttcggggaggtgcggatggggtactacaac gggcacacgaaggtggcggtgaagagcctgaagcagggcagcatgtcccc gaacgccttcctggccgaggccaacctcatgaagcagctgcaacaccagc ggctggttcggctctacgctgtggtcacccaggagcccatctacatcatc actgaatacatggagaatgggagtctagtggattttctcaagaccccttc aggcatcaagttgaccatcaacaaactcctggacatggcagcccaaattg cagaaggcatggcattcattgaagagcggaattattttcatcgtgacctt cgggctgccaacattctggtgtctgacaccctgagctgcaagattgcaga ctttggcctagcacgcctcattgaggacaacgagtacacagccagggagg gggccaagtttcccattaagtggacagcgccagaagccattaactacggg acattcaccatcaagtcagatgtgtggtcttttgggatcctgctgacgga aattgtcacccacggccgcatcccttacccagggatgaccaacccggagg tgattcagaacctggagcgaggctaccgcatggtgcgccctgacaactgt ccagaggagctgtaccaactcatgaggctgtgctggaaggagcgcccaga ggaccggcccacctttgactacctgcgcagtgtgctggaggacttcttca cggccacagagggccagtaccagcctcagccaagctttctagaacaaaaa ctcatctcagaagaggatctgaatagcgccgtcgaccatcatcatcatca tcattgagttaaacgtctccagctaagtaatatgcccg Conclusion: hLck, T7 shows W260R, D283N and E317D. BGH shows D283N, W260R, I361F, mismatched from aa 133 to 228 (approx).