>HTL_56~3 sequence exported from edited chromas file AGGGGGAGCGATCTATATCGCAATTGTGTAGCTATATTCTCAGCGGCGCCGACGAAAGCA AAAAGGAAATTACTGCGGGCAGATAAACGCCGAATTTACAACTCGAAACTTTGCTTTGTC AAAGATATGTACGTAAGCATATATATCCGGATTGTATAATATTCATAAAATAGTAAAGTA TATTTTCTTTGCGCGGTGTTATAATTTGCTCTTTTGCATTTTTCCGTTTTTTTCTTACCT TCTTATCCCCGACAATGGTAATAGTGGTATATTATATAAAGAGGGAAGGCCTCGAGGATA ATAGAAGGCAATGCACCATCAATTGCTTTACCCCTGGTCCGCGACAAAAAAGACACACTG CGAGCTTTCGAACCTCAGATGCTAATATTACGTGTTATATATACCAAACTTTATAAAATG ACATAGATATTTTATGCTGTGATAGCTTTCCTGTTATGGAGAAGCTCTTCTTATTCCCCC TGTCAACTTTCATACTCTTGTAGAATTtCCTTTATGATAGGTTTATCGCTTACGAATTTA GACTTTGATGTGATGGGTTTGGCACCTGtTCTTTTTTCACACCTTTTGCGTGCCTCATCA >HTL_56~4 rev compl sequence exported from edited chromas file CGAGGGGGTGGGGGGGAAGATTCTTTGCGCGGGAAAAAGAGTTGCTCTTttGCAtTTTTC CGTTTTTTTCTTACCTTCTTATCCCCGACAATGGTAATAGTGGTATATTATATAAAGAGG GAAGGCCTCGAGGATAATAGAAGGCAATGCACCATCAATTGCTTTACCCCTGGTCCGCGA CAAAAAAGACACACTGCGAGCTTTCGAACCTCAGATGCTAATATTACGTGTTATATATAC CAAACTTTATAAAATGACATAGATATTTTATGCTGTGATAGCTTTCCTGTTATGGAGAAG CTCTTCTTATTCCCCCTGTCAACTTTCATACTCTTGTAGAATTTCCTTTATGATAGGTTT ATCGCTTACGAATTTAGACTTTGATGTGATGGGTTTGGCACCTGTTCTTTTCCACAACCT TTGCGTGCCCTCAATCAAAAGGGGT >HTL_56~3 edited sub-sequence for alignment TTGCTCTTTTGCATTTTTCCGTTTTTTTCTTACCT TCTTATCCCCGACAATGGTAATAGTGGTATATTATATAAAGAGGGAAGGCCTCGAGGATA ATAGAAGGCAATGCACCATCAATTGCTTTACCCCTGGTCCGCGACAAAAAAGACACACTG CGAGCTTTCGAACCTCAGATGCTAATATTACGTGTTATATATACCAAACTTTATAAAATG ACATAGATATTTTATGCTGTGATAGCTTTCCTGTTATGGAGAAGCTCTTCTTATTCCCCC TGTCAACTTTCATACTCTTGTAGAATTtCCTTTATGATAGGTTTATCGCTTACGAATTTA GACTTTGATGTGATGGGTTTGGCACCTGtTCTTTT >HTL_56~4 rev compl edited sub-sequence for alignment TTGCTCTTttGCAtTTTTC CGTTTTTTTCTTACCTTCTTATCCCCGACAATGGTAATAGTGGTATATTATATAAAGAGG GAAGGCCTCGAGGATAATAGAAGGCAATGCACCATCAATTGCTTTACCCCTGGTCCGCGA CAAAAAAGACACACTGCGAGCTTTCGAACCTCAGATGCTAATATTACGTGTTATATATAC CAAACTTTATAAAATGACATAGATATTTTATGCTGTGATAGCTTTCCTGTTATGGAGAAG CTCTTCTTATTCCCCCTGTCAACTTTCATACTCTTGTAGAATTTCCTTTATGATAGGTTT ATCGCTTACGAATTTAGACTTTGATGTGATGGGTTTGGCACCTGTTCTTTT >Expected in-frame deletion for YHR163W GTAGAATTGCCAACTACAGGATACGGAATGGCGACTCTATATCGCAATTGTGTAGCTATATTCTCAGCGGCGCCGACGAAAGCAAAAAGGAAATTACTACGGGCAGATAAACGCCGAATTTACAACTCGAAACTTTGCTTTGTCAAAGATATGTACGTAAGCATATATATCCGGATTGTATAATATTCATAAAATAGTAAAGTATATTTTCTTTGCGCGGTGTTATAATTTGCTCTTTTGCATTTTTCCGTTTTTTTCTTACCTTTTTATCCCCGACAATGGTAATAGTGGTATATTATATAAAGAGGAAAGGCCTCGAGGATAATAGAAGGCAATGCACCATCAATTGCTTTACCCCTGGTCCGCGACAAAAAGACACACATGCGAGCTTTCGAACCTCAGATGCTAATATTACGTGTTATATATACCAAACTTTATAAAATGACATAGATATTTTATGCTGTGATAGCTTTCCTGTTATGGAGAAGCTCTTCTTATTCCCCCTGTCAACTTTCATACTCTTGTAGAATTTCCTTTATGATAGGTTTATCGCTTACGAATTTAGACTTTGATGTGATGGGTTTGGCACCTGTTCTTTTTCCACAACCTTTGCGTGCCTCATCAATAGCGTTTGATCTGTCGGGAAATTTGTATTTGTA Aligned sequences (ClustalW at http://www.ebi.ac.uk/Tools/msa/clustalw2/) HTL_56~4 CGAGGGGGTGGGGGGGAAGATTCTTTGCGCGGGAAAAAGAGTTGCTCTTTTGCATTTTTC 60 HTL_56~3 -----------------------------------------TTGCTCTTTTGCATTTTTC 19 ******************* HTL_56~4 CGTTTTTTTCTTACCTTCTTATCCCCGACAATGGTAATAGTGGTATATTATATAAAGAGG 120 HTL_56~3 CGTTTTTTTCTTACCTTCTTATCCCCGACAATGGTAATAGTGGTATATTATATAAAGAGG 79 ************************************************************ HTL_56~4 GAAGGCCTCGAGGATAATAGAAGGCAATGCACCATCAATTGCTTTACCCCTGGTCCGCGA 180 HTL_56~3 GAAGGCCTCGAGGATAATAGAAGGCAATGCACCATCAATTGCTTTACCCCTGGTCCGCGA 139 ************************************************************ HTL_56~4 CAAAAAAGACACACTGCGAGCTTTCGAACCTCAGATGCTAATATTACGTGTTATATATAC 240 HTL_56~3 CAAAAAAGACACACTGCGAGCTTTCGAACCTCAGATGCTAATATTACGTGTTATATATAC 199 ************************************************************ HTL_56~4 CAAACTTTATAAAATGACATAGATATTTTATGCTGTGATAGCTTTCCTGTTATGGAGAAG 300 HTL_56~3 CAAACTTTATAAAATGACATAGATATTTTATGCTGTGATAGCTTTCCTGTTATGGAGAAG 259 ************************************************************ HTL_56~4 CTCTTCTTATTCCCCCTGTCAACTTTCATACTCTTGTAGAATTTCCTTTATGATAGGTTT 360 HTL_56~3 CTCTTCTTATTCCCCCTGTCAACTTTCATACTCTTGTAGAATTTCCTTTATGATAGGTTT 319 ************************************************************ HTL_56~4 ATCGCTTACGAATTTAGACTTTGATGTGATGGGTTTGGCACCTGTTCTTTTCCACAACCT 420 HTL_56~3 ATCGCTTACGAATTTAGACTTTGATGTGATGGGTTTGGCACCTGTTCTTTT--------- 370 *************************************************** HTL_56~4 TTGCGTGCCCTCAATCAAAAGGGGT 445 HTL_56~3 ------------------------- Alignment of PCR product from DYHR163W strain with the expected product DYHR163W ------------------------------------------------------------ Expected GTAGAATTGCCAACTACAGGATACGGAATGGCGACTCTATATCGCAATTGTGTAGCTATA 60 DYHR163W ------------------------------------------------------------ Expected TTCTCAGCGGCGCCGACGAAAGCAAAAAGGAAATTACTACGGGCAGATAAACGCCGAATT 120 DYHR163W ------------------------------------------------------------ Expected TACAACTCGAAACTTTGCTTTGTCAAAGATATGTACGTAAGCATATATATCCGGATTGTA 180 DYHR163W -------------------------------------------------TTGCTCTTTTG 11 Expected TAATATTCATAAAATAGTAAAGTATATTTTCTTTGCGCGGTGTTATAATTTGCTCTTTTG 240 *********** DYHR163W CATTTTTCCGTTTTTTTCTTACCTTCTTATCCCCGACAATGGTAATAGTGGTATATTATA 71 Expected CATTTTTCCGTTTTTTTCTTACCTTTTTATCCCCGACAATGGTAATAGTGGTATATTATA 300 ************************* ********************************** DYHR163W TAAAGAGGGAAGGCCTCGAGGATAATAGAAGGCAATGCACCATCAATTGCTTTACCCCTG 131 Expected TAAAGAGGAAAGGCCTCGAGGATAATAGAAGGCAATGCACCATCAATTGCTTTACCCCTG 360 ******** *************************************************** DYHR163W GTCCGCGACAAAAAAGACACAC-TGCGAGCTTTCGAACCTCAGATGCTAATATTACGTGT 190 Expected GTCCGCGACAAAAA-GACACACATGCGAGCTTTCGAACCTCAGATGCTAATATTACGTGT 419 ************** ******* ************************************* DYHR163W TATATATACCAAACTTTATAAAATGACATAGATATTTTATGCTGTGATAGCTTTCCTGTT 250 Expected TATATATACCAAACTTTATAAAATGACATAGATATTTTATGCTGTGATAGCTTTCCTGTT 479 ************************************************************ DYHR163W ATGGAGAAGCTCTTCTTATTCCCCCTGTCAACTTTCATACTCTTGTAGAATTTCCTTTAT 310 Expected ATGGAGAAGCTCTTCTTATTCCCCCTGTCAACTTTCATACTCTTGTAGAATTTCCTTTAT 539 ************************************************************ DYHR163W GATAGGTTTATCGCTTACGAATTTAGACTTTGATGTGATGGGTTTGGCACCTGTTCTTTT 370 Expected GATAGGTTTATCGCTTACGAATTTAGACTTTGATGTGATGGGTTTGGCACCTGTTCTTTT 599 ************************************************************ DYHR163W ------------------------------------------------------------ Expected TCCACAACCTTTGCGTGCCTCATCAATAGCGTTTGATCTGTCGGGAAATTTGTATTTGTA 659 - No additional A in the chromas traces 2 PCR errors?