Parts_for_cell_cell_communication_lab

advertisement
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
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