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N32 Lck sequence analysis

(Sample_Name = N32LckBGH_18) GGTTACTAGCTGGAGACGTTTAACTCAATGATGATGATGATGATGGTC GACGGCGCTATTCAGATCCTCTTCTG AGATGA GTTTTTGTTCTAGAAAGCTTGGTACCGAGCTCGGATCCACTAGTCCAG TGTGGTGGAATTCCGTCTCCCTGGGAAC CTCC CACTCGTCCTCCCACCACGGCTTCTGGGGCTTCTGGGTCTGGCAGGG GCGGCTCAACCGTGTGCACAGCCCATCTG AAGC ATTGGTGTAATGGCGGACCAGTTCATGCAGGCCGGGAAAAGTGATTC GAGGGGAGATGTAGAAGCCACCGTTGTCC AGAT TACGGATCTTGTAATGTTTCACCACCTCTCCCTGGTTCTGGTCGAAGT CCCGGACCGACAGTGAAAACGATCCCGC GGTG CTCTCGCTCTCCCGGATGAGGAAGGAGCCGTGAGTGTTCCCGGGCG CCAGGAGCTGCCGCTCCGCGTCCTTGCGGC TCAG GTTCTTGAAGAACCAGGGTTCGGGCTCCAGGCTGTTCGCTTTGGCCA CAAAATTGAAGGGGATGAAGCCTTCCTGG CCCG TGGTCAGGGACTGCGCCTTCCACCACTCGCCGCTCTGCTCCAGGATG CGGAGCTGTTCCCCCTTCTCAAAGCCCAG ATCT CCGTCGTGAGAGGGCTCATAGCTGTGCAGAGCGATAACCAGGTTGTC TTGCAGTGGGGAAGCCAGCGGATTGGAGC CTTC GTAGGTAACCAGTGGGTCCCGCACCTCAGAGCCATTTCGGATGAGCA GCGTGCCCTTGCCATCCAGTGGGACTATG GGAT AATGGCAGTTCTCACACACATCGATGTTTTCCATCCAGTCATCTTCCG GGTGTGAGCTGCAGCCACAGCCCATGGT GGCC TCGAGTCTAGAGGGCCCGTTTAAACGCTAGCCAGCTTGGGTCTCCCT ATAGTGAGTCGTATAATTTCGATAGCCAG TAAG CAGTGGGTCTCTAGTAGCAGAGAGCTCTGCTATATAGACCTCCCACC GTACACGCTACGCCATTTGCGTCATGGGG

CGGA GTGTACGACTTTTGAAAGTCCCGTGATTTGTGCAAACAACTCCCATGA CGTCATGGGGTGGAGACTGAAATCCGTG AGTC AACCGCTTTCCCGCCATTGATGTACTGCAAACGCCTACACATGTTGCG ATGACATAAACGTAAGTTATCTGCCAAT AGGA AGTCTAGGCATGTACTGGCATATGGCCGTGCTACGCTAGACCTAGGA CCTGTAGAACATATTGCACGGGGTCTAAT AACA CTACT Reverse complement (5 to 3) agtagtgttattagaccccgtgcaatatgttctacaggtcctaggtctag cgtagcacggccatatgccagtacatgcctagacttcctattggcagata acttacgtttatgtcatcgcaacatgtgtaggcgtttgcagtacatcaat ggcgggaaagcggttgactcacggatttcagtctccaccccatgacgtca tgggagttgtttgcacaaatcacgggactttcaaaagtcgtacactccgc cccatgacgcaaatggcgtagcgtgtacggtgggaggtctatatagcaga gctctctgctactagagacccactgcttactggctatcgaaattatacga ctcactatagggagacccaagctggctagcgtttaaacgggccctctaga ctcgaggccaccatgggctgtggctgcagctcacacccggaagatgactg gatggaaaacatcgatgtgtgtgagaactgccattatcccatagtcccac tggatggcaagggcacgctgctcatccgaaatggctctgaggtgcgggac ccactggttacctacgaaggctccaatccgctggcttccccactgcaaga caacctggttatcgctctgcacagctatgagccctctcacgacggagatc tgggctttgagaagggggaacagctccgcatcctggagcagagcggcgag tggtggaaggcgcagtccctgaccacgggccaggaaggcttcatcccctt caattttgtggccaaagcgaacagcctggagcccgaaccctggttcttca agaacctgagccgcaaggacgcggagcggcagctcctggcgcccgggaac actcacggctccttcctcatccgggagagcgagagcaccgcgggatcgtt ttcactgtcggtccgggacttcgaccagaaccagggagaggtggtgaaac attacaagatccgtaatctggacaacggtggcttctacatctcccctcga atcacttttcccggcctgcatgaactggtccgccattacaccaatgcttc agatgggctgtgcacacggttgagccgcccctgccagacccagaagcccc agaagccgtggtgggaggacgagtgggaggttcccagggagacggaattc caccacactggactagtggatccgagctcggtaccaagctttctagaaca aaaactcatctcagaagaggatctgaatagcgccgtcgaccatcatcatc atcatcattgagttaaacgtctccagctagtaacc

Frame1, 98% identical to original N32

(Sample_Name = N32LckT7_17) ACAGAGTGCTGCGTTTACGGGCCCTCTAGA CTCGAGGCCACCATGGGCTGTGGCTGCAGCTCACACCCGGAAGA TGACTG GATGGAAAACATCGATGTGTGTGAGAACTGCCATTATCCCATAGTCCC ACTGGATGGCAAGGGCACGCTGCTCATC CGAA ATGGCTCTGAGGTGCGGGACCCACTGGTTACCTACGAAGGCTCCAAT CCGCTGGCTTCCCCACTGCAAGACAACCT GGTT ATCGCTCTGCACAGCTATGAGCCCTCTCACGACGGAGATCTGGGCTT TGAGAAGGGGGAACAGCTCCGCATCCTGG AGCA GAGCGGCGAGTGGTGGAAGGCGCAGTCCCTGACCACGGGCCAGGAA GGCTTCATCCCCTTCAATTTTGTGGCCAAA GCGA ACAGCCTGGAGCCCGAACCCTGGTTCTTCAAGAACCTGAGCCGCAAG GACGCGGAGCGGCAGCTCCTGGCGCCCGG GAAC ACTCACGGCTCCTTCCTCATCCGGGAGAGCGAGAGCACCGCGGGATC GTTTTCACTGTCGGTCCGGGACTTCGACC AGAA CCAGGGAGAGGTGGTGAAACATTACAAGATCCGTAATCTGGACAACG GTGGCTTCTACATCTCCCCTCGAATCACT TTTC CCGGCCTGCATGAACTGGTCCGCCATTACACCAATGCTTCAGATGGG CTGTGCACACGGTTGAGCCGCCCCTGCCA GACC CAGAAGCCCCAGAAGCCGTGGTGGGAGGACGAGTGGGAGGTTCCCA GGGAGACGGAATTC CACCACACTGGACTAG TGGA TCCGAGCTCGGTACCAAGCTTTCTAGAACAAAAACTCATCTCAGAAGA GGATCTGAATAGCGCCGTCGACCATCAT CATC

ATCATCATTGAGTTTAAACGGTCTCCAGCTTAAGTTTAAACCGCTGATC AGCCTCGACTGTGCCTTCTAGTGCCAG CCAT CTGTGTTTGCCCTCCCCGTGCTCTGACCCTGAGTGCCACTCCACTGTC TTTCTATAAATGAGAATGCATCGCATGT CTGA GTAGTGTCATCTATCTGGGGGGTGGGGGTGGGGCAGAAGCAAGGGG AAGATGGAAAACAATAGCAGCATGCTGGGA TGCG TGGCTCTTATGCTTCGAGGCGAAGAAACAGCTGGGGCCTAGGATCCC ACCGCCTGTACGCCAATACGCCGGGTGTG GTTA CGCGAGGTGAAGCTCACGTGCACGCCTAGGCCCTCGATTCCTCTTTC CAGATTCCGTTCCTCCGCAGCTATTATAG GGGG GGCT Frame1 ok 98% identical to original N32 Compare T7 and BGH reverse 100% identical in nucleotide sequence and ptn sequence. Protein seq:
>N32Lck-BGH_rev_comp Translated - Frame 1 LEAT MGCGCSSHPEDDWMENIDVCENCHYPIVPLDGKGTLLIRNGSEVRDPLVTYEGSNPLA SPLQDNLV IALHSYEPSHDGDLGFEKGEQLRILEQSGEWWKAQSLTTGQEGFIPFNFVAKANSLEPE PWFFKNLSRKD AERQLLAPGNTHGSFLIRESESTAGSFSLSVRDFDQNQGEVVKHYKIRNLDNGGFYISP RITFPGLHELV RHYTNASDGLCTRLSRPCQTQKPQKPWWEDEWEVPRET EF Compared to original N32Lck: mutation at 57, from P to L.

(Sample_Name = N32LckBGH_18) GGTTACTAGCTGGAGACGTTTAACTCAATGATGATGATGATGATGGT CGACGGCGCTATTCAGATCCTCTTCTG AGATGA GTTTTTGTTCTAGAAAGCTTGGTACCGAGCTCGGATCCACTAGTCCAG TGTGGTGGAATTCCGTCTCCCTGGGAAC CTCC CACTCGTCCTCCCACCACGGCTTCTGGGGCTTCTGGGTCTGGCAGGG GCGGCTCAACCGTGTGCACAGCCCATCTG AAGC ATTGGTGTAATGGCGGACCAGTTCATGCAGGCCGGGAAAAGTGATTC GAGGGGAGATGTAGAAGCCACCGTTGTCC AGAT TACGGATCTTGTAATGTTTCACCACCTCTCCCTGGTTCTGGTCGAAGT CCCGGACCGACAGTGAAAACGATCCCGC GGTG CTCTCGCTCTCCCGGATGAGGAAGGAGCCGTGAGTGTTCCCGGGCG CCAGGAGCTGCCGCTCCGCGTCCTTGCGGC TCAG GTTCTTGAAGAACCAGGGTTCGGGCTCCAGGCTGTTCGCTTTGGCCA CAAAATTGAAGGGGATGAAGCCTTCCTGG CCCG TGGTCAGGGACTGCGCCTTCCACCACTCGCCGCTCTGCTCCAGGATG CGGAGCTGTTCCCCCTTCTCAAAGCCCAG ATCT CCGTCGTGAGAGGGCTCATAGCTGTGCAGAGCGATAACCAGGTTGTC TTGCAGTGGGGAAGCCAGCGGATTGGAGC CTTC GTAGGTAACCAGTGGGTCCCGCACCTCAGAGCCATTTCGGATGAGCA GCGTGCCCTTGCCATCCAGTGGGACTATG GGAT AATGGCAGTTCTCACACACATCGATGTTTTCCATCCAGTCATCTTCCG GGTGTGAGCTGCAGCCACAGCCCATGGT GGCC TCGAGTCTAGAGGGCCCGTTTAAACGCTAGCCAGCTTGGGTCTCCCT ATAGTGAGTCGTATAATTTCGATAGCCAG TAAG CAGTGGGTCTCTAGTAGCAGAGAGCTCTGCTATATAGACCTCCCACC GTACACGCTACGCCATTTGCGTCATGGGG

CGGA GTGTACGACTTTTGAAAGTCCCGTGATTTGTGCAAACAACTCCCATGA CGTCATGGGGTGGAGACTGAAATCCGTG AGTC AACCGCTTTCCCGCCATTGATGTACTGCAAACGCCTACACATGTTGCG ATGACATAAACGTAAGTTATCTGCCAAT AGGA AGTCTAGGCATGTACTGGCATATGGCCGTGCTACGCTAGACCTAGGA CCTGTAGAACATATTGCACGGGGTCTAAT AACA CTACT > (5' --> 3' complementary sequence) - 1285 nucleotides agtagtgttattagaccccgtgcaatatgttctacaggtcctaggtctag cgtagcacggccatatgccagtacatgcctagacttcctattggcagata acttacgtttatgtcatcgcaacatgtgtaggcgtttgcagtacatcaat ggcgggaaagcggttgactcacggatttcagtctccaccccatgacgtca tgggagttgtttgcacaaatcacgggactttcaaaagtcgtacactccgc cccatgacgcaaatggcgtagcgtgtacggtgggaggtctatatagcaga gctctctgctactagagacccactgcttactggctatcgaaattatacga ctcactatagggagacccaagctggctagcgtttaaacgggccctctaga ctcgaggccaccatgggctgtggctgcagctcacacccggaagatgactg gatggaaaacatcgatgtgtgtgagaactgccattatcccatagtcccac tggatggcaagggcacgctgctcatccgaaatggctctgaggtgcgggac ccactggttacctacgaaggctccaatccgctggcttccccactgcaaga caacctggttatcgctctgcacagctatgagccctctcacgacggagatc tgggctttgagaagggggaacagctccgcatcctggagcagagcggcgag tggtggaaggcgcagtccctgaccacgggccaggaaggcttcatcccctt caattttgtggccaaagcgaacagcctggagcccgaaccctggttcttca agaacctgagccgcaaggacgcggagcggcagctcctggcgcccgggaac actcacggctccttcctcatccgggagagcgagagcaccgcgggatcgtt ttcactgtcggtccgggacttcgaccagaaccagggagaggtggtgaaac attacaagatccgtaatctggacaacggtggcttctacatctcccctcga atcacttttcccggcctgcatgaactggtccgccattacaccaatgcttc agatgggctgtgcacacggttgagccgcccctgccagacccagaagcccc agaagccgtggtgggaggacgagtgggaggttcccagggagacggaattc caccacactggactagtggatccgagctcggtaccaagctttctagaaca aaaactcatctcagaagaggatctgaatagcgccgtcgaccatcatcatc atcatcattgagttaaacgtctccagctagtaacc

(Sample_Name = N32LckT7_17) ACAGAGTGCTGCGTTTACGGGCCCTCTAGACTCGAGGCCACCATGG GCTGTGGCTGCAGCTCACACCCGGAAGA TGACTG GATGGAAAACATCGATGTGTGTGAGAACTGCCATTATCCCATAGTCCC ACTGGATGGCAAGGGCACGCTGCTCATC CGAA ATGGCTCTGAGGTGCGGGACCCACTGGTTACCTACGAAGGCTCCAAT CCGCTGGCTTCCCCACTGCAAGACAACCT GGTT ATCGCTCTGCACAGCTATGAGCCCTCTCACGACGGAGATCTGGGCTT TGAGAAGGGGGAACAGCTCCGCATCCTGG AGCA GAGCGGCGAGTGGTGGAAGGCGCAGTCCCTGACCACGGGCCAGGAA GGCTTCATCCCCTTCAATTTTGTGGCCAAA GCGA ACAGCCTGGAGCCCGAACCCTGGTTCTTCAAGAACCTGAGCCGCAAG GACGCGGAGCGGCAGCTCCTGGCGCCCGG GAAC ACTCACGGCTCCTTCCTCATCCGGGAGAGCGAGAGCACCGCGGGATC GTTTTCACTGTCGGTCCGGGACTTCGACC AGAA CCAGGGAGAGGTGGTGAAACATTACAAGATCCGTAATCTGGACAACG GTGGCTTCTACATCTCCCCTCGAATCACT TTTC CCGGCCTGCATGAACTGGTCCGCCATTACACCAATGCTTCAGATGGG CTGTGCACACGGTTGAGCCGCCCCTGCCA GACC CAGAAGCCCCAGAAGCCGTGGTGGGAGGACGAGTGGGAGGTTCCCA GGGAGACGGAATTCCACCACACTGGACTAG TGGA TCCGAGCTCGGTACCAAGCTTTCTAGAACAAAAACTCATCTCAGAAGA GGATCTGAATAGCGCCGTCGACCATCAT CATC ATCATCATTGAGTTTAAACGGTCTCCAGCTTAAGTTTAAACCGCTGATC AGCCTCGACTGTGCCTTCTAGTGCCAG CCAT CTGTGTTTGCCCTCCCCGTGCTCTGACCCTGAGTGCCACTCCACTGTC TTTCTATAAATGAGAATGCATCGCATGT CTGA GTAGTGTCATCTATCTGGGGGGTGGGGGTGGGGCAGAAGCAAGGGG

AAGATGGAAAACAATAGCAGCATGCTGGGA TGCG TGGCTCTTATGCTTCGAGGCGAAGAAACAGCTGGGGCCTAGGATCCC ACCGCCTGTACGCCAATACGCCGGGTGTG GTTA CGCGAGGTGAAGCTCACGTGCACGCCTAGGCCCTCGATTCCTCTTTC CAGATTCCGTTCCTCCGCAGCTATTATAG GGGG GGCT Conclusion: hLck, P57L mutation