Kinase domain Lck sequence analysis

(Sample_Name = KinaseLckBGH_20) GACTGTTACTTAAGCTGGAGACGTTTAACTCAATGATGATGATGATG ATGGTCGACGGCGCTATTCAGATCCTC TTCTGA GATGAGTTTTTGTTCTAGAAAGCTTGGTACCGAGCTCGGATCCACTAG TCCAGTGTGGTGGAATTCAGGCTGAGGC TGGT ACTGGCCCTCTGTGGCCGTGAAGAAGTCCTCCAGCACACTGCGCAGG TAGTCAAAGGTGGGCCGGTCCTCTGGGCG CTCC TTCCAGCACAGCCTCATGAGTTGGTACAGCTCCTCTGGACAGTTGTCA GGGCGCACCATGCGGTAGCCTCGCTCCA GGTT CTGAATCACCTCCGGGTTGGTCATCCCTGGGTAAGGGATGCGGCCGT GGGTGACAATTTCCGTCAGCAGGATCCCA AAAG ACCACACATCTGACTTGATGGTGAATGTCCCGTAGTTAATGGCTTCTG GCGCTGTCCACTTAATGGGAAACTTGGC CCCC TCCCTGGCTGTGTACTCGTTGTCCTCAATGAGGCGTGCTAGGCCAAA GTCTGCAATCTTGCAGCTCAGGGTGTCAG ACAC CAGAATGTTGGCAGCCCGAAGGTCACGATGAATATAATTCCGCTCTTC AATGAATGCCATGCCTTCTGCAATTTGG GCTG CCATGTCCAGGAGTTTGTTGATGGTCAACTTGATGCCTGAAGGGGTCT TGAGAAAATCCACTAGACTCCCATTCTC CATG TATTCAGTGATGATGTAGATGGGCTCCTGGGTGACCACAGCGTAGAG CCGAACCAGCCGCTGGTGTTGCAGCTGCT TCAT GAGGTTGGCCTCGGCCAGGAAGGCGTTCGGGGACATGCTGCCCTGC TTCAGGCTCTTCACCGCCACCTTCGTGTGC CCGT TGTAGTACCCCATCCACACCTCCCCGAACTGTCCAGCCCCAGCCGCTC CACCAGCTTCAGTCCCATGTGGCCTCGA GTCT AGAGGGCCCGTTTAAACGCTAGCAGCTGGGGTCTCCTATAGTGAGTC

GTATATTTCGATAGCAGTAGCAGTGGGTC TCTA GTAGCAGAGAGCTCTGCTTATAAGACCTCCACCGTACACGCCTACCG CCATTGCGTCATGGGGCGGATGTACGACT TGGA GTCCGTGATTTGGTGCAAACAACTCCATGGGCGTCATGGGGTGGGAC TGATTCCTGAGTCACCGTTTCACGCCATG ATGT ACTGCACGTACATGTTACGCGCGGTACTACTGATTGCATGGAGTCTTA GCGTATGGGATATGCCAGGGCCTACTCT GTGG TGCATAG Reverse complement of BGH (5 to 3) ctatgcaccacagagtaggccctggcatatcccatacgctaagactccat gcaatcagtagtaccgcgcgtaacatgtacgtgcagtacatcatggcgtg aaacggtgactcaggaatcagtcccaccccatgacgcccatggagttgtt tgcaccaaatcacggactccaagtcgtacatccgccccatgacgcaatgg cggtaggcgtgtacggtggaggtcttataagcagagctctctgctactag agacccactgctactgctatcgaaatatacgactcactataggagacccc agctgctagcgtttaaacgggccctctaga ctcgaggccacatgggactg aagctggtggagcggctggggctggacagttcggggaggtgtggatgggg tactacaacgggcacacgaaggtggcggtgaagagcctgaagcagggcag catgtccccgaacgccttcctggccgaggccaacctcatgaagcagctgc aacaccagcggctggttcggctctacgctgtggtcacccaggagcccatc tacatcatcactgaatacatggagaatgggagtctagtggattttctcaa gaccccttcaggcatcaagttgaccatcaacaaactcctggacatggcag cccaaattgcagaaggcatggcattcattgaagagcggaattatattcat cgtgaccttcgggctgccaacattctggtgtctgacaccctgagctgcaa gattgcagactttggcctagcacgcctcattgaggacaacgagtacacag ccagggagggggccaagtttcccattaagtggacagcgccagaagccatt aactacgggacattcaccatcaagtcagatgtgtggtcttttgggatcct gctgacggaaattgtcacccacggccgcatcccttacccagggatgacca acccggaggtgattcagaacctggagcgaggctaccgcatggtgcgccct gacaactgtccagaggagctgtaccaactcatgaggctgtgctggaagga gcgcccagaggaccggcccacctttgactacctgcgcagtgtgctggagg acttcttcacggccacagagggccagtaccagcctcagcctgaattc cac cacactggactagtggatccgagctcggtaccaagctttctagaacaaaa

actcatctcagaagaggatctgaatagcgccgtcgaccatcatcatcatc atcattgagttaaacgtctccagcttaagtaacagtc Kinase BGH: Frame 2

>Kinase_Lck_-BGH_seq_analysis Translated - Frame 2 SRPHGTEAGGAAGAGQFGEVWMGYYNGHTKVAVKSLKQGSMSPNAFLAEANLMKQL QHQRLVRLYAVVTQ EPIYIITEYMENGSLVDFLKTPSGIKLTINKLLDMAAQIAEGMAFIEERNYIHRDLRAANILV SDTLSCK IADFGLARLIEDNEYTAREGAKFPIKWTAPEAINYGTFTIKSDVWSFGILLTEIVTHGRIPYP GMTNPEV IQNLERGYRMVRPDNCPEELYQLMRLCWKERPEDRPTFDYLRSVLEDFFTATEGQYQP QPEF

Frameshift. Ntd sequence compared to original has 2 gaps, 99% identical. (Sample_Name = KinaseLckT7_19) CCAGGCTGCTGCGTTTACGGGCCCTCTAGA CTCGAGGCCACCATGGGACTGAAGCTGGTGGAGCGGCTGGGGGC TGGACA GTTCGGGGAGGTGTGGATGGGGTACTACAACGGGCACACGAAGGTG GCGGTGAAGAGCCTGAAGCAGGGCAGCATG TCCC CGAACGCCTTCCTGGCCGAGGCCAACCTCATGAAGCAGCTGCAACAC CAGCGGCTGGTTCGGCTCTACGCTGTGGT CACC CAGGAGCCCATCTACATCATCACTGAATACATGGAGAATGGGAGTCT AGTGGATTTTCTCAAGACCCCTTCAGGCA TCAA GTTGACCATCAACAAACTCCTGGACATGGCAGCCCAAATTGCAGAAG GCATGGCATTCATTGAAGAGCGGAATTAT ATTC ATCGTGACCTTCGGGCTGCCAACATTCTGGTGTCTGACACCCTGAGCT GCAAGATTGCAGACTTTGGCCTAGCACG CCTC ATTGAGGACAACGAGTACACAGCCAGGGAGGGGGCCAAGTTTCCCAT TAAGTGGACAGCGCCAGAAGCCATTAACT ACGG GACATTCACCATCAAGTCAGATGTGTGGTCTTTTGGGATCCTGCTGAC GGAAATTGTCACCCACGGCCGCATCCCT TACC

CAGGGATGACCAACCCGGAGGTGATTCAGAACCTGGAGCGAGGCTA CCGCATGGTGCGCCCTGACAACTGTCCAGA GGAG CTGTACCAACTCATGAGGCTGTGCTGGAAGGAGCGCCCAGAGGACC GGCCCACCTTTGACTACCTGCGCAGTGTGC TGGA GGACTTCTTCACGGCCACAGAGGGCCAGTACCAGCCTCAGCCTGAAT TC CACCACACTGGACTAGTGGATCCGAGC TCGG TACCAAGCTTTCTAGAACAAAAACTCATCTCAGAGAGGATCTGAATAG CGCCGTCGACCATCATCATCATCATCAT TGAG TTTAAACGGTCTCCAGCTTAAGTTTAAACCGCTGATCAGCTCGACTGT GCTTCTAGTTGCAGCATCTGTGTTGCCC CTCC CCGTGCTCTGACCTGAGTGCACTCCCACTGTCTTTCTATAAATGAGAA TGCATCGCATGTCTGATAGGTGTCATTT ATTC TGGGGGGTGGGGTGGGCAGACGCAGGGAGGATTGGGAGACATACA GCATGCTGGATGCGTTGCCCTATGCTCCGAG GCGA GACCCTGGCTAAGGAATCCAGCTGTAGCTAGCGCGGTAGGTACCGAG GTAGCTCATGCAGCCAGGCCGCCCTCGT T7 seq: frame1
>kinaseLck_-_T7_seq_results Translated - Frame 1 LEATMGLKLVERLGAGQFGEVWMGYYNGHTKVAVKSLKQGSMSPNAFLAEANLMKQL QHQRLVRLYAVVT QEPIYIITEYMENGSLVDFLKTPSGIKLTINKLLDMAAQIAEGMAFIEERNYIHRDLRAANIL VSDTLSC KIADFGLARLIEDNEYTAREGAKFPIKWTAPEAINYGTFTIKSDVWSFGILLTEIVTHGRIPY PGMTNPE VIQNLERGYRMVRPDNCPEELYQLMRLCWKERPEDRPTFDYLRSVLEDFFTATEGQYQ PQPEF Nucleotide sequence compared to the original is 99% identical- has two mutations in ntd seq. Protein sequence is 99% similar to original, D to N mutation at T7 and BGH nucleotide comparison 99% similar, 2 gaps. Ptn comparison of T7, BGH

100% similar from aa 13 onwards

(Sample_Name = KinaseLckBGH_20) GACTGTTACTTAAGCTGGAGACGTTTAACTCAATGATGATGATGATG ATGGTCGACGGCGCTATTCAGATCCTC TTCTGA GATGAGTTTTTGTTCTAGAAAGCTTGGTACCGAGCTCGGATCCACTAG TCCAGTGTGGTGGAATTCAGGCTGAGGC TGGT ACTGGCCCTCTGTGGCCGTGAAGAAGTCCTCCAGCACACTGCGCAGG TAGTCAAAGGTGGGCCGGTCCTCTGGGCG CTCC TTCCAGCACAGCCTCATGAGTTGGTACAGCTCCTCTGGACAGTTGTCA GGGCGCACCATGCGGTAGCCTCGCTCCA GGTT CTGAATCACCTCCGGGTTGGTCATCCCTGGGTAAGGGATGCGGCCGT GGGTGACAATTTCCGTCAGCAGGATCCCA AAAG ACCACACATCTGACTTGATGGTGAATGTCCCGTAGTTAATGGCTTCTG GCGCTGTCCACTTAATGGGAAACTTGGC CCCC TCCCTGGCTGTGTACTCGTTGTCCTCAATGAGGCGTGCTAGGCCAAA GTCTGCAATCTTGCAGCTCAGGGTGTCAG ACAC CAGAATGTTGGCAGCCCGAAGGTCACGATGAATATAATTCCGCTCTTC AATGAATGCCATGCCTTCTGCAATTTGG GCTG CCATGTCCAGGAGTTTGTTGATGGTCAACTTGATGCCTGAAGGGGTCT TGAGAAAATCCACTAGACTCCCATTCTC CATG TATTCAGTGATGATGTAGATGGGCTCCTGGGTGACCACAGCGTAGAG CCGAACCAGCCGCTGGTGTTGCAGCTGCT TCAT GAGGTTGGCCTCGGCCAGGAAGGCGTTCGGGGACATGCTGCCCTGC TTCAGGCTCTTCACCGCCACCTTCGTGTGC CCGT TGTAGTACCCCATCCACACCTCCCCGAACTGTCCAGCCCCAGCCGCTC CACCAGCTTCAGTCCCATGTGGCCTCGA GTCT

AGAGGGCCCGTTTAAACGCTAGCAGCTGGGGTCTCCTATAGTGAGTC GTATATTTCGATAGCAGTAGCAGTGGGTC TCTA GTAGCAGAGAGCTCTGCTTATAAGACCTCCACCGTACACGCCTACCG CCATTGCGTCATGGGGCGGATGTACGACT TGGA GTCCGTGATTTGGTGCAAACAACTCCATGGGCGTCATGGGGTGGGAC TGATTCCTGAGTCACCGTTTCACGCCATG ATGT ACTGCACGTACATGTTACGCGCGGTACTACTGATTGCATGGAGTCTTA GCGTATGGGATATGCCAGGGCCTACTCT GTGG TGCATAG > (5' --> 3' complementary sequence) - 1287 nucleotides ctatgcaccacagagtaggccctggcatatcccatacgctaagactccat gcaatcagtagtaccgcgcgtaacatgtacgtgcagtacatcatggcgtg aaacggtgactcaggaatcagtcccaccccatgacgcccatggagttgtt tgcaccaaatcacggactccaagtcgtacatccgccccatgacgcaatgg cggtaggcgtgtacggtggaggtcttataagcagagctctctgctactag agacccactgctactgctatcgaaatatacgactcactataggagacccc agctgctagcgtttaaacgggccctctagactcgaggccacatgggactg aagctggtggagcggctggggctggacagttcggggaggtgtggatgggg tactacaacgggcacacgaaggtggcggtgaagagcctgaagcagggcag catgtccccgaacgccttcctggccgaggccaacctcatgaagcagctgc aacaccagcggctggttcggctctacgctgtggtcacccaggagcccatc tacatcatcactgaatacatggagaatgggagtctagtggattttctcaa gaccccttcaggcatcaagttgaccatcaacaaactcctggacatggcag cccaaattgcagaaggcatggcattcattgaagagcggaattatattcat cgtgaccttcgggctgccaacattctggtgtctgacaccctgagctgcaa gattgcagactttggcctagcacgcctcattgaggacaacgagtacacag ccagggagggggccaagtttcccattaagtggacagcgccagaagccatt aactacgggacattcaccatcaagtcagatgtgtggtcttttgggatcct gctgacggaaattgtcacccacggccgcatcccttacccagggatgacca acccggaggtgattcagaacctggagcgaggctaccgcatggtgcgccct gacaactgtccagaggagctgtaccaactcatgaggctgtgctggaagga gcgcccagaggaccggcccacctttgactacctgcgcagtgtgctggagg acttcttcacggccacagagggccagtaccagcctcagcctgaattccac cacactggactagtggatccgagctcggtaccaagctttctagaacaaaa actcatctcagaagaggatctgaatagcgccgtcgaccatcatcatcatc atcattgagttaaacgtctccagcttaagtaacagtc

(Sample_Name = KinaseLckT7_19) CCAGGCTGCTGCGTTTACGGGCCCTCTAGACTCGAGGCCACCATGG GACTGAAGCTGGTGGAGCGGCTGGGGGC TGGACA GTTCGGGGAGGTGTGGATGGGGTACTACAACGGGCACACGAAGGTG GCGGTGAAGAGCCTGAAGCAGGGCAGCATG TCCC CGAACGCCTTCCTGGCCGAGGCCAACCTCATGAAGCAGCTGCAACAC CAGCGGCTGGTTCGGCTCTACGCTGTGGT CACC CAGGAGCCCATCTACATCATCACTGAATACATGGAGAATGGGAGTCT AGTGGATTTTCTCAAGACCCCTTCAGGCA TCAA GTTGACCATCAACAAACTCCTGGACATGGCAGCCCAAATTGCAGAAG GCATGGCATTCATTGAAGAGCGGAATTAT ATTC ATCGTGACCTTCGGGCTGCCAACATTCTGGTGTCTGACACCCTGAGCT GCAAGATTGCAGACTTTGGCCTAGCACG CCTC ATTGAGGACAACGAGTACACAGCCAGGGAGGGGGCCAAGTTTCCCAT TAAGTGGACAGCGCCAGAAGCCATTAACT ACGG GACATTCACCATCAAGTCAGATGTGTGGTCTTTTGGGATCCTGCTGAC GGAAATTGTCACCCACGGCCGCATCCCT TACC CAGGGATGACCAACCCGGAGGTGATTCAGAACCTGGAGCGAGGCTA CCGCATGGTGCGCCCTGACAACTGTCCAGA GGAG CTGTACCAACTCATGAGGCTGTGCTGGAAGGAGCGCCCAGAGGACC GGCCCACCTTTGACTACCTGCGCAGTGTGC TGGA GGACTTCTTCACGGCCACAGAGGGCCAGTACCAGCCTCAGCCTGAAT TCCACCACACTGGACTAGTGGATCCGAGC TCGG TACCAAGCTTTCTAGAACAAAAACTCATCTCAGAGAGGATCTGAATAG CGCCGTCGACCATCATCATCATCATCAT TGAG TTTAAACGGTCTCCAGCTTAAGTTTAAACCGCTGATCAGCTCGACTGT GCTTCTAGTTGCAGCATCTGTGTTGCCC

CTCC CCGTGCTCTGACCTGAGTGCACTCCCACTGTCTTTCTATAAATGAGAA TGCATCGCATGTCTGATAGGTGTCATTT ATTC TGGGGGGTGGGGTGGGCAGACGCAGGGAGGATTGGGAGACATACA GCATGCTGGATGCGTTGCCCTATGCTCCGAG GCGA GACCCTGGCTAAGGAATCCAGCTGTAGCTAGCGCGGTAGGTACCGAG GTAGCTCATGCAGCCAGGCCGCCCTCGT

LKLVERLGAGQFGEVWMGYYNGHTKVAVKSLKQGSMSP+AFLAEANLMKQLQHQRLVRLY Conclusion: hLck, D283N mutation