Supplementary Data 2 1. Correct construct for EFE expression on the genome of Synechococcus elongatus strain GD-EFE7942 & EFE7942 1 – 890, S. elongatus 7942 NS1 homologous region A 1770 – 2561, Spectinomycin resistance gene ORF, aadA1 2976 – 3036, PpsbA promoter 3037 – 3071, RBSv4 3072 – 4190, EFE-FLAG ORF 4267 – 4314, T7 terminator 4412 – 5425, S. elongatus 7942 NS1 homologous region B CGGCAGCCGGCGGAGCGCTGCTTTCTTGGCAAGCGGTCGCCAGCCCCAACGCCAGGGCTGCCAGCCCGAAACAGCGGGGCAAGGCAGCTTGGAAGGGCGATCGCAGCACGGGCATGGCAATGTCTCTCTGAAGGAATGCAGACCTTATTCGTACAGCCAGGGTTGAATCGTGGGGGTCCAATCACTTAGCTCTGCTGGGCTAAACCAGAGAGCAATTTCCTGTTGTGCTGTTTCGATTGCATCCGAGCCATGGATGATGTTGCGGCCAATATTGACACCAAAATCACCACGGATGGTGCCCGGTTCTGCCGTCAGCGGATTGGTAGCGCCGATCAACTTGCGAGCAGCCGCCACAACGCCTTCGCCTTCCAAGACGATCGCCACGATCGGCCCAGAGGTGATGAACTCGACGAGGCCATTGAAGAAGGGGCGCTCGCGGTGGACAGCATAGTGCTGTTCGGCCAGCTCGCGACTGGGCTTCAGCTGCTTTAGGCCCACCAGTTTGAAGCCTTTTTGCTCAAAGCGGCCGATGATCGTACCGACCAAACCCCGCTGAACGCCATCGGGCTTGATGGCAATAAATGTGCGTTCCACAGACATCTAGATAGTCCTCAAGACGAGGCAAGCATTGAGCTTGCCTTCCTATGGTTCGGGATCACTGGGATTCTTGACAAGCGATCGCGGTCACATCGCTATCTCTTAGGACTTCGCAGCGGGCGAGTCGGATTGACCCGGTAGGGATTTCGCCAGATCAATGCCCGTGGTTTGTTTCAGCTTCTCCAGCAAGCTAGCGATTTGGGTAGCGCTGCCTTCCCCTTCGCCAATCACAGTGATCGACTCCACGTCGATATCTGGCACGGTGCCTGAAAGCGTGACGAGCAGGGACTCGAAGCTTGCATGCCTGCAGGTCGACTCTAGAGCTTTATGCTTGTAAACCGTTTTGTGAAAAAATTTTTAAAATAAAAAAGGGGACCTCTAGGGTCCCCAATTAATTAGTAATATAATCTATTAAAGGTCATTCAAAAGGTCATCCACCGGATCAATTCCCCTGCTCGCGCAGGCTGGGTGCCAAGCTCTCGGGTAACATCAAGGCCCGATCCTTGGAGCCCTTGCCCTCCCGCACGATGATCGTGCCGTGATCGAAATCCAGATCCTTGACCCGCAGTTGCAAACCCTCACTGATCCGCATGCCCGTTCCATACAGAAGCTGGGCGAACAAACGATGCTCGCCTTCCAGAAAACCGAGGATGCGAACCACTTCATCCGGGGTCAGCACCACCGGCAAGCGCCGCGACGGCCGAGGTCTTCCGATCTCCTGAAGCCAGGGCAGATCCGTGCACAGCACCTTGCCGTAGAAGAACAGCAAGGCCGCCAATGCCTGACGATGCGTGGAGACCGAAACCTTGCGCTCGTTCGCCAGCCAGGACAGAAATGCCTCGACTTCGCTGCTGCCCAAGGTTGCCGGGTGACGCACACCGTGGAAACGGATGAAGGCACGAACCCAGTGGACATAAGCCTGTTCGGTTCGTAAGCTGTAATGCAAGTAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGGGGTACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAAACATCATGAGGGAAGCGGTGATCGCCGAAGTATCGACTCAACTATCAGAGGTAGTTGGCGTCATCGAGCGCCATCTCGAACCGACGTTGCTGGCCGTACATTTGTACGGCTCCGCAGTGGATGGCGGCCTGAAGCCACACAGTGATATTGATTTGCTGGTTACGGTGACCGTAAGGCTTGATGAAACAACGCGGCGAGCTTTGATCAACGACCTTTTGGAAACTTCGGCTTCCCCTGGAGAGAGCGAGATTCTCCGCGCTGTAGAAGTCACCATTGTTGTGCACGACGACATCATTCCGTGGCGTTATCCAGCTAAGCGCGAACTGCAATTTGGAGAATGGCAGCGCAATGACATTCTTGCAGGTATCTTCGAGCCAGCCACGATCGACATTGATCTGGCTATCTTGCTGACAAAAGCAAGAGAACATAGCGTTGCCTTGGTAGGTCCAGCGGCGGAGGAACTCTTTGATCCGGTTCCTGAACAGGATCTATTTGAGGCGCTAAATGAAACCTTAACGCTATGGAACTCGCCGCCCGACTGGGCTGGCGATGAGCGAAATGTAGTGCTTACGTTGTCCCGCATTTGGTACAGCGCAGTAACCGGCAAAATCGCGCCGAAGGATGTCGCTGCCGACTGGGCAATGGAGCGCCTGCCGGCCCAGTATCAGCCCGTCATACTTGAAGCTAGACAGGCTTATCTTGGACAAGAAGAAGATCGCTTGGCCTCGCGCGCAGATCAGTTGGAAGAATTTGTCCACTACGTGAAAGGCGAGATCACCAAGGTAGTCGGCAAATAATGTCTAACAATTCGTTCAAGCCGACGCCGCTTCGCGGCGCG GCTTAACTCAAGCGTTAGATGCACTAAGCACATAATTGCTCACAGCCAAACTATCAGGTCAAGTCTGCTTTTATTATTTTTAAGCGTGCATAATAAGCCCTACACAAATTGGGAGATATATCATGAAAGGCTGGCTTTTTCTTGTTATCGCAATAGTTGGCGAAGTAATCGCAACATCCGCATTAAAATCTAGCGAGGGCTTTACTAAGCTGATCCGGTGGATGACCTTTTGAATGACCTTTAATAGATTATATTACTAATTAATTGGGGACCCTAGAGGTCCCCTTTTTTATTTTAAAAATTTTTTCACAAAACGGTTTACAAGCATAAAGCTCTAGAGGATCGGCGGCCGCGGATCCCCTGAGGGGAATTCGATCTCAATGAATATTGGTTGACACGGGCGTATAAGACATGTTATACTGTTGAATAACAAGTTTACCGTTCCCAAAAATAAAGAAGGAGGAACAGCATGACCAATTTGCAAACTTTTGAATTACCCACCGAAGTGACTGGCTGTGCCGCTGATATTTCCTTAGGTCGCGCCCTGATTCAAGCCTGGCAGAAAGACGGCATTTTTCAAATTAAAACCGATAGTGAACAAGACCGTAAAACTCAAGAAGCTATGGCCGCTAGCAAACAGTTTTGTAAAGAACCCTTGACCTTTAAATCCAGTTGCGTGAGCGATTTAACTTATTCTGGCTACGTGGCCTCCGGTGAAGAAGTTACCGCTGGGAAACCCGACTTTCCCGAAATTTTTACCGTGTGTAAAGATTTATCCGTGGGCGACCAACGGGTTAAAGCTGGATGGCCCTGTCATGGCCCCGTTCCCTGGCCCAACAACACCTACCAGAAAAGTATGAAAACTTTTATGGAAGAATTGGGGTTAGCCGGAGAACGCTTGTTAAAACTGACCGCTTTGGGGTTTGAACTGCCCATTAATACCTTTACTGATTTGACCCGTGACGGATGGCATCACATGCGCGTGTTACGTTTTCCCCCCCAAACCTCCACTCTGAGTCGGGGCATTGGTGCCCATACCGATTATGGTCTGTTGGTGATTGCCGCTCAGGATGACGTTGGCGGTCTGTACATTCGTCCCCCCGTGGAAGGGGAAAAACGGAATCGCAACTGGTTGCCCGGCGAAAGCTCTGCCGGCATGTTTGAACATGACGAACCCTGGACCTTTGTTACCCCCACTCCCGGGGTGTGGACCGTTTTTCCCGGAGATATTCTGCAATTTATGACCGGGGGACAGTTACTGTCCACTCCCCATAAAGTGAAATTGAATACCCGTGAACGGTTTGCCTGTGCTTATTTTCACGAACCCAACTTTGAAGCCTCTGCTTACCCCTTGTTTGAACCCTCCGCCAATGAACGGATTCATTATGGCGAACACTTTACCAACATGTTTATGCGGTGCTACCCCGATCGCATTACCACTCAACGTATTAACAAAGAAAACCGGTTAGCCCACCTGGAAGATTTGAAAAAATACAGTGACACTCGCGCTACTGGTAGCGATTACAAAGATCATGATGGCGATTACAAAGATCACGACATCGATTACAAAGATGATGATGATAAATAAGAATTCGAAGCTTGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCTCGAGGGTACCGAGCTCGAATTTCGAGCTTCTGGAGCAGGAAGATGTCGCGGGCATTAGCACCAGCGGTCTGCCAAGCCTCCGCCAGCCGTTGGGTCCCTTCCGCTTGAGCTTTTCCATCTTCGACGATACGGGCGGCGGCCCCCCGCGCTTCCGCGATCGCCCGTTTACAAGCTGCCTCAGCTGGGGCGATCACATCGGCTTGAAGTTGCTGCTGCACCTGTTTGATCCGCTCCTGCTGCACAGGGAGTTCTGCTTGGCTACGAGCGACTTCGGTAGCAATGTCCGCTTCAGCTTCGGCCACCACCGCTTCGCGCCGCGTCAACGCATCCTGAATCCGGCGCTCGGCCTCGGCTTGGGCGATCGCTACATCGCGATCGATCCGACGCAGGGCCGTGATCTTGTCATTTTCGGCCGTTTGGATCGCAGAGGCAGCCTGGGCATCGGCTTCAGCAATTCGGGCATCTCGCTGCAGATCAGCCCGCTGCTTGCGTCCACTAGCCGAGAGATAACCGACCTCATCGGAAATGTTCTGGACTTGCAGCGTATCGAGGACTAGACCCAGCTGCTCAAGGTCATCCTCCGCCTCTTCCAGCAGACTTTTGGCAAAGGCAATTTTGTCCTCGTTGATCTGCTCCGGCGTGAGGCTGGCTAAAACACCACGCAAGTTGCCTTCGAGGGTCTCCTTGGCAATTTGCTCGATTTCCTTACGGTTTTTGCCAAGCAGCCGCTCGATCGCGTTGTGGATGGTCGGTTCTTCCCCAGCAATCTTGATATTGGCAACGCCTTCAACAGTCAGGGGAATGCCGCCCTTGGAGAAGGCATTGGAAACGCGCAACTCAATGATCATGTTGGTCAGATCCATGCGGAGCGCTTTTTCCAGCAGAGGTACCCGCAGGCTGCTGCCGCCCTTGACCAAGCGATAGCCAACTCGGCGGCCATCACTACTGCGGCGACTACTGCCAGCAAAGATCAAAATTTCACTGGGTTGGCAGATGTAGTAGAGATTGCGCAGGACTAAGCTGCCAGCCCCGGCG 2. Mutation Type 1: Fragment 2603-4709 (2107 bp) is missing from EFE7942 colonies #1,2, 4, 8, 9. 1 – 890, S. elongatus 7942 NS1 homologous region A 1770 – 2561, Spectinomycin resistance gene ORF, aadA1 2603 – 3318, S. elongatus 7942 NS1 homologous region B, truncated due to spontaneous mutation CGGCAGCCGGCGGAGCGCTGCTTTCTTGGCAAGCGGTCGCCAGCCCCAACGCCAGGGCTGCCAGCCCGAAACAGCGGGGCAAGGCAGCTTGGAAGGGCGATCGCAGCACGGGCATGGCAATGTCTCTCTGAAGGAATGCAGACCTTATTCGTACAGCCAGGGTTGAATCGTGGGGGTCCAATCACTTAGCTCTGCTGGGCTAAACCAGAGAGCAATTTCCTGTTGTGCTGTTTCGATTGCATCCGAGCCATGGATGATGTTGCGGCCAATATTGACACCAAAATCACCACGGATGGTGCCCGGTTCTGCCGTCAGCGGATTGGTAGCGCCGATCAACTTGCGAGCAGCCGCCACAACGCCTTCGCCTTCCAAGACGATCGCCACGATCGGCCCAGAGGTGATGAACTCGACGAGGCCATTGAAGAAGGGGCGCTCGCGGTGGACAGCATAGTGCTGTTCGGCCAGCTCGCGACTGGGCTTCAGCTGCTTTAGGCCCACCAGTTTGAAGCCTTTTTGCTCAAAGCGGCCGATGATCGTACCGACCAAACCCCGCTGAACGCCATCGGGCTTGATGGCAATAAATGTGCGTTCCACAGACATCTAGATAGTCCTCAAGACGAGGCAAGCATTGAGCTTGCCTTCCTATGGTTCGGGATCACTGGGATTCTTGACAAGCGATCGCGGTCACATCGCTATCTCTTAGGACTTCGCAGCGGGCGAGTCGGATTGACCCGGTAGGGATTTCGCCAGATCAATGCCCGTGGTTTGTTTCAGCTTCTCCAGCAAGCTAGCGATTTGGGTAGCGCTGCCTTCCCCTTCGCCAATCACAGTGATCGACTCCACGTCGATATCTGGCACGGTGCCTGAAAGCGTGACGAGCAGGGACTCGAAGCTTGCATGCCTGCAGGTCGACTCTAGAGCTTTATGCTTGTAAACCGTTTTGTGAAAAAATTTTTAAAATAAAAAAGGGGACCTCTAGGGTCCCCAATTAATTAGTAATATAATCTATTAAAGGTCATTCAAAAGGTCATCCACCGGATCAATTCCCCTGCTCGCGCAGGCTGGGTGCCAAGCTCTCGGGTAACATCAAGGCCCGATCCTTGGAGCCCTTGCCCTCCCGCACGATGATCGTGCCGTGATCGAAATCCAGATCCTTGACCCGCAGTTGCAAACCCTCACTGATCCGCATGCCCGTTCCATACAGAAGCTGGGCGAACAAACGATGCTCGCCTTCCAGAAAACCGAGGATGCGAACCACTTCATCCGGGGTCAGCACCACCGGCAAGCGCCGCGACGGCCGAGGTCTTCCGATCTCCTGAAGCCAGGGCAGATCCGTGCACAGCACCTTGCCGTAGAAGAACAGCAAGGCCGCCAATGCCTGACGATGCGTGGAGACCGAAACCTTGCGCTCGTTCGCCAGCCAGGACAGAAATGCCTCGACTTCGCTGCTGCCCAAGGTTGCCGGGTGACGCACACCGTGGAAACGGATGAAGGCACGAACCCAGTGGACATAAGCCTGTTCGGTTCGTAAGCTGTAATGCAAGTAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGGGGTACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAAACATCATGAGGGAAGCGGTGATCGCCGAAGTATCGACTCAACTATCAGAGGTAGTTGGCGTCATCGAGCGCCATCTCGAACCGACGTTGCTGGCCGTACATTTGTACGGCTCCGCAGTGGATGGCGGCCTGAAGCCACACAGTGATATTGATTTGCTGGTTACGGTGACCGTAAGGCTTGATGAAACAACGCGGCGAGCTTTGATCAACGACCTTTTGGAAACTTCGGCTTCCCCTGGAGAGAGCGAGATTCTCCGCGCTGTAGAAGTCACCATTGTTGTGCACGACGACATCATTCCGTGGCGTTATCCAGCTAAGCGCGAACTGCAATTTGGAGAATGGCAGCGCAATGACATTCTTGCAGGTATCTTCGAGCCAGCCACGATCGACATTGATCTGGCTATCTTGCTGACAAAAGCAAGAGAACATAGCGTTGCCTTGGTAGGTCCAGCGGCGGAGGAACTCTTTGATCCGGTTCCTGAACAGGATCTATTTGAGGCGCTAAATGAAACCTTAACGCTATGGAACTCGCCGCCCGACTGGGCTGGCGATGAGCGAAATGTAGTGCTTACGTTGTCCCGCATTTGGTACAGCGCAGTAACCGGCAAAATCGCGCCGAAGGATGTCGCTGCCGACTGGGCAATGGAGCGCCTGCCGGCCCAGTATCAGCCCGTCATACTTGAAGCTAGACAGGCTTATCTTGGACAAGAAGAAGATCGCTTGGCCTCGCGCGCAGATCAGTTGGAAGAATTTGTCCACTACGTGAAAGGCGAGATCACCAAGGTAGTCGGCAAATAATGTCTAACAATTCGTTCAAGCCGACGCCGCTTCGCGGCGCG TCAACGCATCCTGAATCCGGCGCTCGGCCTCGGCTTGGGCGATCGCTACATCGCGATCGATCCGACGCAGGGCCGTGATCTTGTCATTTTCGGCCGTTTGGATCGCAGAGGCAGCCTGGGCATCGGCTTCAGCAATTCGGGCATCTCGCTGCAGATCAGCCCGCTGCTTGCGTCCACTAGCCGAGAGATAACCGACCTCATCGGAAATGTTCTGGACTTGCAGCGTATCGAGGACTAGACCCAGCTGCTCAAGGTCATCCTCCGCCTCTTCCAGCAGACTTTTGGCAAAGGCAATTTTGTCCTCGTTGATCTGCTCCGGCGTGAGGCTGGCTAAAACACCACGCAAGTTGCCTTCGAGGGTCTCCTTGGCAATTTGCTCGATTTCCTTACGGTTTTTGCCAAGCAGCCGCTCGATCGCGTTGTGGATGGTCGGTTCTTCCCCAGCAATCTTGATATTGGCAACGCCTTCAACAGTCAGGGGAATGCCGCCCTTGGAGAAGGCATTGGAAACGCGCAACTCAATGATCATGTTGGTCAGATCCATGCGGAGCGCTTTTTCCAGCAGAGGTACCCGCAGGCTGCTGCCGCCCTTGACCAAGCGATAGCCAACTCGGCGGCCATCACTACTGCGGCGACTACTGCCAGCAAAGATCAAAATTTCACTGGGTTGGCAGATGTAGTAGAGATTGCGCAGGACTAAGCTGCCAGCCCCGGCG 3. Mutation Type 2: Fragment 3940-3963 (24 bp) is missing from EFE7942 colonies #3, 5, 7, 10. 1 – 890, S. elongatus 7942 NS1 homologous region A 1770 – 2561, Spectinomycin resistance gene ORF, aadA1 2976 – 3036, PpsbA promoter 3037 – 3071, RBSv4 3072 – 4166, EFE-FLAG ORF, 24 bp within the original ORF deleted due to spontaneous mutation 4243 – 4290, T7 terminator 4388 – 5401, S. elongatus 7942 NS1 homologous region B CGGCAGCCGGCGGAGCGCTGCTTTCTTGGCAAGCGGTCGCCAGCCCCAACGCCAGGGCTGCCAGCCCGAAACAGCGGGGCAAGGCAGCTTGGAAGGGCGATCGCAGCACGGGCATGGCAATGTCTCTCTGAAGGAATGCAGACCTTATTCGTACAGCCAGGGTTGAATCGTGGGGGTCCAATCACTTAGCTCTGCTGGGCTAAACCAGAGAGCAATTTCCTGTTGTGCTGTTTCGATTGCATCCGAGCCATGGATGATGTTGCGGCCAATATTGACACCAAAATCACCACGGATGGTGCCCGGTTCTGCCGTCAGCGGATTGGTAGCGCCGATCAACTTGCGAGCAGCCGCCACAACGCCTTCGCCTTCCAAGACGATCGCCACGATCGGCCCAGAGGTGATGAACTCGACGAGGCCATTGAAGAAGGGGCGCTCGCGGTGGACAGCATAGTGCTGTTCGGCCAGCTCGCGACTGGGCTTCAGCTGCTTTAGGCCCACCAGTTTGAAGCCTTTTTGCTCAAAGCGGCCGATGATCGTACCGACCAAACCCCGCTGAACGCCATCGGGCTTGATGGCAATAAATGTGCGTTCCACAGACATCTAGATAGTCCTCAAGACGAGGCAAGCATTGAGCTTGCCTTCCTATGGTTCGGGATCACTGGGATTCTTGACAAGCGATCGCGGTCACATCGCTATCTCTTAGGACTTCGCAGCGGGCGAGTCGGATTGACCCGGTAGGGATTTCGCCAGATCAATGCCCGTGGTTTGTTTCAGCTTCTCCAGCAAGCTAGCGATTTGGGTAGCGCTGCCTTCCCCTTCGCCAATCACAGTGATCGACTCCACGTCGATATCTGGCACGGTGCCTGAAAGCGTGACGAGCAGGGACTCGAAGCTTGCATGCCTGCAGGTCGACTCTAGAGCTTTATGCTTGTAAACCGTTTTGTGAAAAAATTTTTAAAATAAAAAAGGGGACCTCTAGGGTCCCCAATTAATTAGTAATATAATCTATTAAAGGTCATTCAAAAGGTCATCCACCGGATCAATTCCCCTGCTCGCGCAGGCTGGGTGCCAAGCTCTCGGGTAACATCAAGGCCCGATCCTTGGAGCCCTTGCCCTCCCGCACGATGATCGTGCCGTGATCGAAATCCAGATCCTTGACCCGCAGTTGCAAACCCTCACTGATCCGCATGCCCGTTCCATACAGAAGCTGGGCGAACAAACGATGCTCGCCTTCCAGAAAACCGAGGATGCGAACCACTTCATCCGGGGTCAGCACCACCGGCAAGCGCCGCGACGGCCGAGGTCTTCCGATCTCCTGAAGCCAGGGCAGATCCGTGCACAGCACCTTGCCGTAGAAGAACAGCAAGGCCGCCAATGCCTGACGATGCGTGGAGACCGAAACCTTGCGCTCGTTCGCCAGCCAGGACAGAAATGCCTCGACTTCGCTGCTGCCCAAGGTTGCCGGGTGACGCACACCGTGGAAACGGATGAAGGCACGAACCCAGTGGACATAAGCCTGTTCGGTTCGTAAGCTGTAATGCAAGTAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGGGGTACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAAACATCATGAGGGAAGCGGTGATCGCCGAAGTATCGACTCAACTATCAGAGGTAGTTGGCGTCATCGAGCGCCATCTCGAACCGACGTTGCTGGCCGTACATTTGTACGGCTCCGCAGTGGATGGCGGCCTGAAGCCACACAGTGATATTGATTTGCTGGTTACGGTGACCGTAAGGCTTGATGAAACAACGCGGCGAGCTTTGATCAACGACCTTTTGGAAACTTCGGCTTCCCCTGGAGAGAGCGAGATTCTCCGCGCTGTAGAAGTCACCATTGTTGTGCACGACGACATCATTCCGTGGCGTTATCCAGCTAAGCGCGAACTGCAATTTGGAGAATGGCAGCGCAATGACATTCTTGCAGGTATCTTCGAGCCAGCCACGATCGACATTGATCTGGCTATCTTGCTGACAAAAGCAAGAGAACATAGCGTTGCCTTGGTAGGTCCAGCGGCGGAGGAACTCTTTGATCCGGTTCCTGAACAGGATCTATTTGAGGCGCTAAATGAAACCTTAACGCTATGGAACTCGCCGCCCGACTGGGCTGGCGATGAGCGAAATGTAGTGCTTACGTTGTCCCGCATTTGGTACAGCGCAGTAACCGGCAAAATCGCGCCGAAGGATGTCGCTGCCGACTGGGCAATGGAGCGCCTGCCGGCCCAGTATCAGCCCGTCATACTTGAAGCTAGACAGGCTTATCTTGGACAAGAAGAAGATCGCTTGGCCTCGCGCGCAGATCAGTTGGAAGAATTTGTCCACTACGTGAAAGGCGAGATCACCAAGGTAGTCGGCAAATAATGTCTAACAATTCGTTCAAGCCGACGCCGCTTCGCGGCGCG GCTTAACTCAAGCGTTAGATGCACTAAGCACATAATTGCTCACAGCCAAACTATCAGGTCAAGTCTGCTTTTATTATTTTTAAGCGTGCATAATAAGCCCTACACAAATTGGGAGATATATCATGAAAGGCTGGCTTTTTCTTGTTATCGCAATAGTTGGCGAAGTAATCGCAACATCCGCATTAAAATCTAGCGAGGGCTTTACTAAGCTGATCCGGTGGATGACCTTTTGAATGACCTTTAATAGATTATATTACTAATTAATTGGGGACCCTAGAGGTCCCCTTTTTTATTTTAAAAATTTTTTCACAAAACGGTTTACAAGCATAAAGCTCTAGAGGATCGGCGGCCGCGGATCCCCTGAGGGGAATTCGATCTCAATGAATATTGGTTGACACGGGCGTATAAGACATGTTATACTGTTGAATAACAAGTTTACCGTTCCCAAAAATAAAGAAGGAGGAACAGCATGACCAATTTGCAAACTTTTGAATTACCCACCGAAGTGACTGGCTGTGCCGCTGATATTTCCTTAGGTCGCGCCCTGATTCAAGCCTGGCAGAAAGACGGCATTTTTCAAATTAAAACCGATAGTGAACAAGACCGTAAAACTCAAGAAGCTATGGCCGCTAGCAAACAGTTTTGTAAAGAACCCTTGACCTTTAAATCCAGTTGCGTGAGCGATTTAACTTATTCTGGCTACGTGGCCTCCGGTGAAGAAGTTACCGCTGGGAAACCCGACTTTCCCGAAATTTTTACCGTGTGTAAAGATTTATCCGTGGGCGACCAACGGGTTAAAGCTGGATGGCCCTGTCATGGCCCCGTTCCCTGGCCCAACAACACCTACCAGAAAAGTATGAAAACTTTTATGGAAGAATTGGGGTTAGCCGGAGAACGCTTGTTAAAACTGACCGCTTTGGGGTTTGAACTGCCCATTAATACCTTTACTGATTTGACCCGTGACGGATGGCATCACATGCGCGTGTTACGTTTTCCCCCCCAAACCTCCACTCTGAGTCGGGGCATTGGTGCCCATACCGATTATGGTCTGTTGGTGATTGCCGCTCAGGATGACGTTGGCGGTCTGTACATTCGTCCCCCCGTGGAAGGGGAAAAACGGAATCGCAACTGGTTGCCCGGCGAAAGCTCTGCCGGCATGTTTGAACATGACGAACCCTGGACCTTTGTTACCCCCACTCCCGGGGTGTGGACCGTTTTTCCCGGAGATATTCTGCAATTTATGACCGGGGGACAGTTACTGTCCACTCCCCATAAAGTGAAATTGAATACCCGTGAACGGTTTGCCTGTGCTTATTTTCACGAACCCAACTTTGAAGCCTCCGCCAATGAACGGATTCATTATGGCGAACACTTTACCAACATGTTTATGCGGTGCTACCCCGATCGCATTACCACTCAACGTATTAACAAAGAAAACCGGTTAGCCCACCTGGAAGATTTGAAAAAATACAGTGACACTCGCGCTACTGGTAGCGATTACAAAGATCATGATGGCGATTACAAAGATCACGACATCGATTACAAAGATGATGATGATAAATAAGAATTCGAAGCTTGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCTCGAGGGTACCGAGCTCGAATTTCGAGCTTCTGGAGCAGGAAGATGTCGCGGGCATTAGCACCAGCGGTCTGCCAAGCCTCCGCCAGCCGTTGGGTCCCTTCCGCTTGAGCTTTTCCATCTTCGACGATACGGGCGGCGGCCCCCCGCGCTTCCGCGATCGCCCGTTTACAAGCTGCCTCAGCTGGGGCGATCACATCGGCTTGAAGTTGCTGCTGCACCTGTTTGATCCGCTCCTGCTGCACAGGGAGTTCTGCTTGGCTACGAGCGACTTCGGTAGCAATGTCCGCTTCAGCTTCGGCCACCA CCGCTTCGCGCCGCGTCAACGCATCCTGAATCCGGCGCTCGGCCTCGGCTTGGGCGATCGCTACATCGCGATCGATCCGACGCAGGGCCGTGATCTTGTCATTTTCGGCCGTTTGGATCGCAGAGGCAGCCTGGGCATCGGCTTCAGCAATTCGGGCATCTCGCTGCAGATCAGCCCGCTGCTTGCGTCCACTAGCCGAGAGATAACCGACCTCATCGGAAATGTTCTGGACTTGCAGCGTATCGAGGACTAGACCCAGCTGCTCAAGGTCATCCTCCGCCTCTTCCAGCAGACTTTTGGCAAAGGCAATTTTGTCCTCGTTGATCTGCTCCGGCGTGAGGCTGGCTAAAACACCACGCAAGTTGCCTTCGAGGGTCTCCTTGGCAATTTGCTCGATTTCCTTACGGTTTTTGCCAAGCAGCCGCTCGATCGCGTTGTGGATGGTCGGTTCTTCCCCAGCAATCTTGATATTGGCAACGCCTTCAACAGTCAGGGGAATGCCGCCCTTGGAGAAGGCATTGGAAACGCGCAACTCAATGATCATGTTGGTCAGATCCATGCGGAGCGCTTTTTCCAGCAGAGGTACCCGCAGGCTGCTGCCGCCCTTGACCAAGCGATAGCCAACTCGGCGGCCATCACTACTGCGGCGACTACTGCCAGCAAAGATCAAAATTTCACTGGGTTGGCAGATGTAGTAGAGATTGCGCAGGACTAAGCTGCCAGCCCCGGCG 4. Mutation Type 3: Fragment 3603 (1 bp) is missing from EFE7942 colony #6. 1 – 890, S. elongatus 7942 NS1 homologous region A 1770 – 2561, Spectinomycin resistance gene ORF, aadA1 2976 – 3036, PpsbA promoter 3037 – 3071, RBSv4 3072 – 4189, EFE-FLAG ORF, the 532th bp within the original ORF deleted due to spontaneous mutation 4266 – 4313, T7 terminator 4411 – 5424, S. elongatus 7942 NS1 homologous region B CGGCAGCCGGCGGAGCGCTGCTTTCTTGGCAAGCGGTCGCCAGCCCCAACGCCAGGGCTGCCAGCCCGAAACAGCGGGGCAAGGCAGCTTGGAAGGGCGATCGCAGCACGGGCATGGCAATGTCTCTCTGAAGGAATGCAGACCTTATTCGTACAGCCAGGGTTGAATCGTGGGGGTCCAATCACTTAGCTCTGCTGGGCTAAACCAGAGAGCAATTTCCTGTTGTGCTGTTTCGATTGCATCCGAGCCATGGATGATGTTGCGGCCAATATTGACACCAAAATCACCACGGATGGTGCCCGGTTCTGCCGTCAGCGGATTGGTAGCGCCGATCAACTTGCGAGCAGCCGCCACAACGCCTTCGCCTTCCAAGACGATCGCCACGATCGGCCCAGAGGTGATGAACTCGACGAGGCCATTGAAGAAGGGGCGCTCGCGGTGGACAGCATAGTGCTGTTCGGCCAGCTCGCGACTGGGCTTCAGCTGCTTTAGGCCCACCAGTTTGAAGCCTTTTTGCTCAAAGCGGCCGATGATCGTACCGACCAAACCCCGCTGAACGCCATCGGGCTTGATGGCAATAAATGTGCGTTCCACAGACATCTAGATAGTCCTCAAGACGAGGCAAGCATTGAGCTTGCCTTCCTATGGTTCGGGATCACTGGGATTCTTGACAAGCGATCGCGGTCACATCGCTATCTCTTAGGACTTCGCAGCGGGCGAGTCGGATTGACCCGGTAGGGATTTCGCCAGATCAATGCCCGTGGTTTGTTTCAGCTTCTCCAGCAAGCTAGCGATTTGGGTAGCGCTGCCTTCCCCTTCGCCAATCACAGTGATCGACTCCACGTCGATATCTGGCACGGTGCCTGAAAGCGTGACGAGCAGGGACTCGAAGCTTGCATGCCTGCAGGTCGACTCTAGAGCTTTATGCTTGTAAACCGTTTTGTGAAAAAATTTTTAAAATAAAAAAGGGGACCTCTAGGGTCCCCAATTAATTAGTAATATAATCTATTAAAGGTCATTCAAAAGGTCATCCACCGGATCAATTCCCCTGCTCGCGCAGGCTGGGTGCCAAGCTCTCGGGTAACATCAAGGCCCGATCCTTGGAGCCCTTGCCCTCCCGCACGATGATCGTGCCGTGATCGAAATCCAGATCCTTGACCCGCAGTTGCAAACCCTCACTGATCCGCATGCCCGTTCCATACAGAAGCTGGGCGAACAAACGATGCTCGCCTTCCAGAAAACCGAGGATGCGAACCACTTCATCCGGGGTCAGCACCACCGGCAAGCGCCGCGACGGCCGAGGTCTTCCGATCTCCTGAAGCCAGGGCAGATCCGTGCACAGCACCTTGCCGTAGAAGAACAGCAAGGCCGCCAATGCCTGACGATGCGTGGAGACCGAAACCTTGCGCTCGTTCGCCAGCCAGGACAGAAATGCCTCGACTTCGCTGCTGCCCAAGGTTGCCGGGTGACGCACACCGTGGAAACGGATGAAGGCACGAACCCAGTGGACATAAGCCTGTTCGGTTCGTAAGCTGTAATGCAAGTAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGGGGTACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAAACATCATGAGGGAAGCGGTGATCGCCGAAGTATCGACTCAACTATCAGAGGTAGTTGGCGTCATCGAGCGCCATCTCGAACCGACGTTGCTGGCCGTACATTTGTACGGCTCCGCAGTGGATGGCGGCCTGAAGCCACACAGTGATATTGATTTGCTGGTTACGGTGACCGTAAGGCTTGATGAAACAACGCGGCGAGCTTTGATCAACGACCTTTTGGAAACTTCGGCTTCCCCTGGAGAGAGCGAGATTCTCCGCGCTGTAGAAGTCACCATTGTTGTGCACGACGACATCATTCCGTGGCGTTATCCAGCTAAGCGCGAACTGCAATTTGGAGAATGGCAGCGCAATGACATTCTTGCAGGTATCTTCGAGCCAGCCACGATCGACATTGATCTGGCTATCTTGCTGACAAAAGCAAGAGAACATAGCGTTGCCTTGGTAGGTCCAGCGGCGGAGGAACTCTTTGATCCGGTTCCTGAACAGGATCTATTTGAGGCGCTAAATGAAACCTTAACGCTATGGAACTCGCCGCCCGACTGGGCTGGCGATGAGCGAAATGTAGTGCTTACGTTGTCCCGCATTTGGTACAGCGCAGTAACCGGCAAAATCGCGCCGAAGGATGTCGCTGCCGACTGGGCAATGGAGCGCCTGCCGGCCCAGTATCAGCCCGTCATACTTGAAGCTAGACAGGCTTATCTTGGACAAGAAGAAGATCGCTTGGCCTCGCGCGCAGATCAGTTGGAAGAATTTGTCCACTACGTGAAAGGCGAGATCACCAAGGTAGTCGGCAAATAATGTCTAACAATTCGTTCAAGCCGACGCCGCTTCGCGGCGCG GCTTAACTCAAGCGTTAGATGCACTAAGCACATAATTGCTCACAGCCAAACTATCAGGTCAAGTCTGCTTTTATTATTTTTAAGCGTGCATAATAAGCCCTACACAAATTGGGAGATATATCATGAAAGGCTGGCTTTTTCTTGTTATCGCAATAGTTGGCGAAGTAATCGCAACATCCGCATTAAAATCTAGCGAGGGCTTTACTAAGCTGATCCGGTGGATGACCTTTTGAATGACCTTTAATAGATTATATTACTAATTAATTGGGGACCCTAGAGGTCCCCTTTTTTATTTTAAAAATTTTTTCACAAAACGGTTTACAAGCATAAAGCTCTAGAGGATCGGCGGCCGCGGATCCCCTGAGGGGAATTCGATCTCAATGAATATTGGTTGACACGGGCGTATAAGACATGTTATACTGTTGAATAACAAGTTTACCGTTCCCAAAAATAAAGAAGGAGGAACAGCATGACCAATTTGCAAACTTTTGAATTACCCACCGAAGTGACTGGCTGTGCCGCTGATATTTCCTTAGGTCGCGCCCTGATTCAAGCCTGGCAGAAAGACGGCATTTTTCAAATTAAAACCGATAGTGAACAAGACCGTAAAACTCAAGAAGCTATGGCCGCTAGCAAACAGTTTTGTAAAGAACCCTTGACCTTTAAATCCAGTTGCGTGAGCGATTTAACTTATTCTGGCTACGTGGCCTCCGGTGAAGAAGTTACCGCTGGGAAACCCGACTTTCCCGAAATTTTTACCGTGTGTAAAGATTTATCCGTGGGCGACCAACGGGTTAAAGCTGGATGGCCCTGTCATGGCCCCGTTCCCTGGCCCAACAACACCTACCAGAAAAGTATGAAAACTTTTATGGAAGAATTGGGGTTAGCCGGAGAACGCTTGTTAAAACTGACCGCTTTGGGGTTTGAACTGCCCATTAATACCTTTACTGATTTGACCCGTGACGGATGGCATCACATGCGCGTGTTACGTTTTCCCCCCAAACCTCCACTCTGAGTCGGGGCATTGGTGCCCATACCGATTATGGTCTGTTGGTGATTGCCGCTCAGGATGACGTTGGCGGTCTGTACATTCGTCCCCCCGTGGAAGGGGAAAAACGGAATCGCAACTGGTTGCCCGGCGAAAGCTCTGCCGGCATGTTTGAACATGACGAACCCTGGACCTTTGTTACCCCCACTCCCGGGGTGTGGACCGTTTTTCCCGGAGATATTCTGCAATTTATGACCGGGGGACAGTTACTGTCCACTCCCCATAAAGTGAAATTGAATACCCGTGAACGGTTTGCCTGTGCTTATTTTCACGAACCCAACTTTGAAGCCTCTGCTTACCCCTTGTTTGAACCCTCCGCCAATGAACGGATTCATTATGGCGAACACTTTACCAACATGTTTATGCGGTGCTACCCCGATCGCATTACCACTCAACGTATTAACAAAGAAAACCGGTTAGCCCACCTGGAAGATTTGAAAAAATACAGTGACACTCGCGCTACTGGTAGCGATTACAAAGATCATGATGGCGATTACAAAGATCACGACATCGATTACAAAGATGATGATGATAAATAAGAATTCGAAGCTTGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCTCGAGGGTACCGAGCTCGAATTTCGAGCTTCTGGAGCAGGAAGATGTCGCGGGCATTAGCACCAGCGGTCTGCCAAGCCTCCGCCAGCCGTTGGGTCCCTTCCGCTTGAGCTTTTCCATCTTCGACGATACGGGCGGCGGCCCCCCGCGCTTCCGCGATCGCCCGTTTACAAGCTGCCTCAGCTGGGGCGATCACATCGGCTTGAAGTTGCTGCTGCACCTGTTTGATCCGCTCCTGCTGCACAGGGAGTTCTGCTTGGCTACGAGCGACTTCGGTAGCAATGTCCGCTTCAGCTTCGGCCACCACCGCTTCGCGCCGCGTCAACGCATCCTGAATCCGGCGCTCGGCCTCGGCTTGGGCGATCGCTACATCGCGATCGATCCGACGCAGGGCCGTGATCTTGTCATTTTCGGCCGTTTGGATCGCAGAGGCAGCCTGGGCATCGGCTTCAGCAATTCGGGCATCTCGCTGCAGATCAGCCCGCTGCTTGCGTCCACTAGCCGAGAGATAACCGACCTCATCGGAAATGTTCTGGACTTGCAGCGTATCGAGGACTAGACCCAGCTGCTCAAGGTCATCCTCCGCCTCTTCCAGCAGACTTTTGGCAAAGGCAATTTTGTCCTCGTTGATCTGCTCCGGCGTGAGGCTGGCTAAAACACCACGCAAGTTGCCTTCGAGGGTCTCCTTGGCAATTTGCTCGATTTCCTTACGGTTTTTGCCAAGCAGCCGCTCGATCGCGTTGTGGATGGTCGGTTCTTCCCCAGCAATCTTGATATTGGCAACGCCTTCAACAGTCAGGGGAATGCCGCCCTTGGAGAAGGCATTGGAAACGCGCAACTCAATGATCATGTTGGTCAGATCCATGCGGAGCGCTTTTTCCAGCAGAGGTACCCGCAGGCTGCTGCCGCCCTTGACCAAGCGATAGCCAACTCGGCGGCCATCACTACTGCGGCGACTACTGCCAGCAAAGATCAAAATTTCACTGGGTTGGCAGATGTAGTAGAGATTGCGCAGGACTAAGCTGCCAGCCCCGGCG