Cell-cell communication lab parts: Plasmids with response regulator proteins: Arrangement for expression of R genes: EcoRI XbaI pTet promoter spacer of 8 StrongRBS (BBa_0034) spacer of 6 R gene Double Terminator(BBa_0010-0012) SpeI PstI Plasmid R1 (1028 bp): RhlR (responds to C4HSL) Plasmid R2 (989 bp): LuxR (responds to 3OC6HSL) Plasmid R3 (964 bp): TraR (responds to 3OC8HSL) Plasmid R4 (1118 bp): LasR (responds to 3OC12HSL) Plasmid with test promoters: Arrangement for testing promoter sequences: EcoRI XbaI Double Terminator Test Promoter StrongRBS Gemini Double Terminator SpeI PstI Plasmid P1 (1385 bp) : Plas1 (from PrsaL gene) Plasmid P2 (1383 bp): Plas 2 (from PA0572) Plasmid P3 (1392 bp): Plas 3 (from PA3904) Plasmid P4 (1452 bp): Plas 4 (from PA4677) Plasmid P5 (1405 bp): Plas 5 (from PA1897) Plasmid P6 (1379 bp): Prhl 1 (from PA3479) Plasmid P7 (1491 bp): Prhl 2 (from PA4217) Plasmid P8 (1309 bp): Plux 1 (from PluxI) Plasmid P9 (1405 bp): Ptra 1 (from PtraI) Literature cited: Sequence information for promoters to test came from: Schuster et al.(2004) PNAS vol. 101(45) 15833-15839. Whiteley and Greenberg (2001) J Bacteriol. 183(19): 5529–5534. 1 Plasmids with response regulator proteins: R1: Ptet_rhlR (BBa_K082021) GAATTCTCTAGATccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagagaaagaggagaaatactagatgaggaatgacggag gctttttgctgtggtgggacggtttgcgtagcgagatgcagccgatccacgacagccagggcgtgttcgccgtcctggaaaaggaagtgcggcgcctgggcttcgattactacg cctatggcgtgcgccacacgattcccttcacccggccgaagaccgaggtccatggcacctatcccaaggcctggctggagcgataccagatgcagaactacggggccgtgga tccggcgatcctcaacggcctgcgctcctcggaaatggtggtctggagcgacagcctgttcgaccagagccggatgctctggaacgaggctcgcgattggggcctctgtgtcgg cgcgaccttgccgatccgcgcgccgaacaatttgctcagcgtgctttccgtggcgcgcgaccagcagaacatctccagcttcgagcgcgaggaaatccgcctgcggctgcgttg catgatcgagttgctgacccagaagctgaccgacctggagcatccgatgctgatgtccaacccggtctgcctgagccatcgcgaacgcgagatcctgcaatggaccgccgacg gcaagagttccggggaaatcgccatcatcctgagcatctccgagagcacggtgaacttccaccacaagaacatccagaagaagttcgacgcgccgaacaagacgctggctg ccgcctacgccgcggcgctgggtctcatcgctgcaaacgacgaaaactacgctttagtagcttaataacactgatagtgctagtgtagatcactactagagccaggcatcaaat aaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttataA CTAGTCTGCAG R2: Ptet luxR (LuxR from BBa_C0062) GAATTCTCTAGATccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagagaaagaggagaaatactagatgaaaaacataaatg ccgacgacacatacagaataattaataaaattaaagcttgtagaagcaataatgatattaatcaatgcttatctgatatgactaaaatggtacattgtgaatattatttactcgc gatcatttatcctcattctatggttaaatctgatatttcaatcctagataattaccctaaaaaatggaggcaatattatgatgacgctaatttaataaaatatgatcctatagtaga ttattctaactccaatcattcaccaattaattggaatatatttgaaaacaatgctgtaaataaaaaatctccaaatgtaattaaagaagcgaaaacatcaggtcttatcactggg tttagtttccctattcatacggctaacaatggcttcggaatgcttagttttgcacattcagaaaaagacaactatatagatagtttatttttacatgcgtgtatgaacataccatta attgttccttctctagttgataattatcgaaaaataaatatagcaaataataaatcaaacaacgatttaaccaaaagagaaaaagaatgtttagcgtgggcatgcgaaggaaa aagctcttgggatatttcaaaaatattaggttgcagtgagcgtactgtcactttccatttaaccaatgcgcaaatgaaactcaatacaacaaaccgctgccaaagtatttctaaa gcaattttaacaggagcaattgattgcccatactttaaaaattaataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtc ggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata ACTAGTCTGCAG R3: Ptet traR (TraR BBa_ BBa_K553000) GAATTCTCTAGAtccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagagaaagaggagaaatactagaaagaggagaaatact agatgcagcactggctggacaagctgactgatcttgccgcgatcgaaggcgatgagtgcatcctgaagaccgggctggcggacatcgccgaccatttcggcttcaccggctat gcctaccttcatatccagcacaggcacatcaccgccgttaccaactatcaccgccaatggcaatcaacctacttcgacaagaagttcgaagcgctcgatccggtcgtcaaacg cgcgaggtcccggaagcacatcttcacctggtcgggcgagcacgagcggccgacgctgtcgaaggacgagcgtgccttctatgaccacgcatccgatttcggcatccgctccg gcatcacaatacccatcaagaccgccaacggctttatgtcgatgttcacgatggcatcggacaagccggtgatcgatctcgatcgggagatcgatgcagtcgcagccgctgca accatcgggcagatccatgcccgcatcccattccttcgcaccacccctaccgcggaggatgccgcatggctcgatccgaaggaggccacctatctgagatggattgccgtcgg caagacgatggaggagatcgccgacgtcgaaggggtcaagtacaacagcgtccgcgtcaagctacgcgaagccatgaagcgcttcgacgtccgcagcaaggcccatcttac cgcgctcgCcatccggcggaaactcatctgatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacg ctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttataACTAGTCTGCAG R4: Ptet lasR (LasR BBa_C0079) GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataTccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagagaaagaggagaaatactagatggc cttggttgacggttttcttgagctggaacgctcaagtggaaaattggagtggagcgccatcctccagaagatggcgagcgaccttggattctcgaagatcctgttcggcctgttg cctaaggacagccaggactacgagaacgccttcatcgtcggcaactacccggccgcctggcgcgagcattacgaccgggctggctacgcgcgggtcgacccgacggtcagtc actgtacccagagcgtactgccgattttctgggaaccgtccatctaccagacgcgaaagcagcacgagttcttcgaggaagcctcggccgccggcctggtgtatgggctgacc atgccgctgcatggtgctcgcggcgaactcggcgcgctgagcctcagcgtggaagcggaaaaccgggccgaggccaaccgtttcatagagtcggtcctgccgaccctgtgga tgctcaaggactacgcactgcaaagcggtgccggactggccttcgaacatccggtcagcaaaccggtggttctgaccagccgggagaaggaagtgttgcagtggtgcgccat cggcaagaccagttgggagatatcggttatctgcaactgctcggaagccaatgtgaacttccatatgggaaatattcggcggaagttcggtgtgacctcccgccgcgtagcgg ccattatggccgttaatttgggtcttattactctcgctgcaaacgacgaaaactacgctttagtagcttaataaccaggcatcaaataaaacgaaaggctcagtcgaaagactg ggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttataACTAGTCTGCAG 2 Plasmids with test promoters: P1: (from PrsaL): GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataCTTCACTTCCTCCAAATAGGAAGCTGAAGAATTTATGCAAATTTCATAATTTTATAACTAGCAAATGA GATAGATTTCGGTGAACCCGGACCCTTGCTAGGCTCGAAGAAACGTCCGAAAAGATCAGAGCAaaagaggagaaatactagatgaccatgatt acggattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagag gcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgcgcgggcggcagcgaaggcggcggcagcgaacatcatcatcatcatcatggcagcgaa cgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacg gaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactaccctgacctatggtgttcaatgctttgcgagatacccagatcatatg aaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaagg tgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggc agacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatgg ccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggatt acacatggcatggatgaactatacaaataataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctct actagagtcacactggctcaccttcgggtgggcctttctgcgtttata ACTAGTCTGCAG P2: (from PA0572): GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataCGAACAATCGGTAAAAATAAAATCAAAATAATCAGACGCAGTTCAAAAACAACCAGAACAGATAGT TTTTCCGCCTTTCCCAACATGCTCCTCTCGCCTTGACTCGTTCGATGCAGCGGATCGCCCATGaaagaggagaaatactagatgaccatgattacg gattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggccc gcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgcgcgggcggcagcgaaggcggcggcagcgaacatcatcatcatcatcatggcagcgaacgta aaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaa aacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactaccctgacctatggtgttcaatgctttgcgagatacccagatcatatgaaa cagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtga tacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcaga caaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccct gtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacac atggcatggatgaactatacaaataataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactag agtcacactggctcaccttcgggtgggcctttctgcgtttata ACTAGTCTGCAG P3: (from PA3904): GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataAAATCCCTTCAAGCGCAACCAGGCAAGGACGCGGTTGCCCAAGCCGTTTCCGAAGGTGCCTCCTGA GCAGTTCAGATAGGTTGTCCGACCCCGAGTGGCACGGCAGAGTGCCAGGGTTTATCTCATAGGGACTTGAGCaaagaggagaaatactag atgaccatgattacggattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgta atagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgcgcgggcggcagcgaaggcggcggcagcgaacatcatcatcatcatc atggcagcgaacgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtg atgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactaccctgacctatggtgttcaatgctttgcgagatac ccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagt caagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgt atacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactcc aattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaaca gctgctgggattacacatggcatggatgaactatacaaataataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggt gaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttataACTAGTCTGCAG 3 P4: (from PA4677): GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataGGCTTGCTCCGCTTCCCTGCCGGCAACTAGCTATCACTTGGAGTAGCTGGCTGGCCTTCTGGCACCG CCGTACCCTTACCAGCATTCCGATTCTCGCCTGCATACCGCAGGCAGGTCCCTTCATGCGGTAACCCTTCGCGGGAGTGCGAAGTCAG GTCGACAGCGGCCGGTGGCAGGATGCGCAGACTGGAGCCCCCCaaagaggagaaatactagatgaccatgattacggattcactggccgtcgttttaca acgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaaca gttgcgcagcctgaatggcgaatggcgcgcgggcggcagcgaaggcggcggcagcgaacatcatcatcatcatcatggcagcgaacgtaaaggagaagaacttttcactgg agttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgca ctactggaaaactacctgttccatggccaacacttgtcactaccctgacctatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgcc atgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagtt aaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaa gttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattac ctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaa taataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggt gggcctttctgcgtttataACTAGTCTGCAG P5: (from PA1897): GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataGACAACAAACCTTGGCCCATGGCGGGCCTCGTCGACGAGGCTCCCCGGGGACCAGAAATGGCAAC TACATACTTCCCCCCCTATCTCTCCGCAGATACCTGCCCGGAAGGGCAGGTTGTCCCTGCCGGGCTGTGACAATTTAATTCGACCAaaa gaggagaaatactagatgaccatgattacggattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctt tcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgcgcgggcggcagcgaaggcggcggcagcgaac atcatcatcatcatcatggcagcgaacgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtgga gagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactaccctgacctatggtgttcaat gctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaag acacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactat aactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatc aacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttc ttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttat ctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata ACTAGTCTGCAG P6: (from PA3479): GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataCCCCGCCGGCCGCTCTGGCACGCCTTATCGCGGGCGGGCAGGGGCTTATGCGCAGGCGGCCGCCC GTCCTGTGAAATCTGGCAGTTACCGTTAGCTTTCGAATTGGCTAAAAAGTGTTCATCGGCaaagaggagaaatactagatgaccatgattacgga ttcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgc accgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgcgcgggcggcagcgaaggcggcggcagcgaacatcatcatcatcatcatggcagcgaacgtaaa ggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaa cttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactaccctgacctatggtgttcaatgctttgcgagatacccagatcatatgaaaca gcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgata cccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagaca aacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgt ccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacat ggcatggatgaactatacaaataataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactaga gtcacactggctcaccttcgggtgggcctttctgcgtttata ACTAGTCTGCAG 4 P7:(from PA4217): GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataCGACACCGCTGCGCCGGCGTTTCATGGCGGATAACCGCAAGCGGTTATTCGCCCTACGCGGC CTTGGAGCCCATCTAACCGCACGCGGTCATGCGTACCGCGGCCTCGGAGCCGGTTCGTAGGGCGAATGACGCCACCGGCGT TATCCGCCGCTGCGCCGACGTTTCATCGCGGTAAACGGTCATCCATCCCAGCCGAACCCCCATCGATTCGAACACTCGAGA AAAGGAAGCACCCaaagaggagaaatactagatgaccatgattacggattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaa tcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgcgcgggcggca gcgaaggcggcggcagcgaacatcatcatcatcatcatggcagcgaacgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatg ggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcac taccctgacctatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttc aaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttgg acacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaag cgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaa aagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataaccaggcatcaaataaaacgaaaggctcagtcg aaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata ACTAGTCTGCAG P8: (from PluxI): GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataacctgtaggatcgtacaggtttacgcaagaaaatggtttgttatagtcgaataaaaaagaggagaaatactagatgaccatgatt acggattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagag gcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggcgcgcgggcggcagcgaaggcggcggcagcgaacatcatcatcatcatcatggcagcgaa cgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacg gaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactaccctgacctatggtgttcaatgctttgcgagatacccagatcatatg aaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaagg tgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggc agacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatgg ccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggatt acacatggcatggatgaactatacaaataataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctct actagagtcacactggctcaccttcgggtgggcctttctgcgtttataACTAGTCTGCAG P9: (from PtraI) GAATTCTCTAGAccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc accttcgggtgggcctttctgcgtttataaagtaacgcgaagtgcagatttgcacatgaaatcaacgctgcacgagatcgattgcacgcgcaaatcgctttttgaagactgactc agtagaatcactacgtgcagatctgcacatagccacaccctgaatgagatgttttctctccgctaaaagaggagaaatactagatgaccatgattacggattcactggccgtcg ttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttc ccaacagttgcgcagcctgaatggcgaatggcgcgcgggcggcagcgaaggcggcggcagcgaacatcatcatcatcatcatggcagcgaacgtaaaggagaagaactttt cactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaattt atttgcactactggaaaactacctgttccatggccaacacttgtcactaccctgacctatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaa gagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaa tcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatgg aatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagaca accattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaac tatacaaataataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctca ccttcgggtgggcctttctgcgtttataACTAGTCTGCAG 5