LOCUS       sequence-472144-       12311 bp    DNA     circular SYN 08-MAY-2025
DEFINITION  Optimized thiosulfate sensor with recombinase switch and fluorescent
            reporter (GFP), constitutive mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 42 to 76)
  AUTHORS   Buss MT, Zhu L, Kwon JH, Tabor JJ, Shapiro MG
  TITLE     Probiotic acoustic biosensors for noninvasive imaging of gut 
            inflammation
  JOURNAL   bioRxiv 2024.09.23.614598
REFERENCE   2  (bases 1 to 12311)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.09.23.614598"
COMMENT     Sequence Label: MB 94C thsS(t3)R-Bxb1_P7-sfGFP_mCherry
FEATURES             Location/Qualifiers
     source          1..12311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..76
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             86..97
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             105..815
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(250..268)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     518..537
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        2092..2126
                     /label=BBa_J23104
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.72"
     terminator      3962..4033
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4049..4076
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     4143..4161
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4182..4216
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     terminator      4880..4951
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4967..4994
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             5547..7046
                     /codon_start=1
                     /transl_table=11
                     /gene="Bxb1"
                     /product="Bxb1 integrase"
                     /label=Bxb1 integrase
                     /note="Bxb1 large serine integrase from Mycobacterium 
                     bacteriophage Bxb1 (Ghosh et al. 2003)."
                     /protein_id="AFF61396.1"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     terminator      complement(7424..7453)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     protein_bind    7505..7554
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    7609..7656
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     misc_RNA        7663..7712
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             7770..7781
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7788..8501
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(8524..8541)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      8694..8780
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8872..8899
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        8918..9009
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9467..9922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9554..9573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9764..9785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(10478..11137)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWCEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     11052..11071
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(11138..11240)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(11632..11649)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11766..12311)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(11846..11865)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcatc ttattaatca gataaaatcg ccagatatcg attgacggct agctcagtcc
       61 taggtacagt gctagcgaat tcattaaaga ggagaaaggt acccatggtg agcaagggcg
      121 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
      181 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      241 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
      301 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
      361 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      421 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
      481 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
      541 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
      601 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
      661 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
      721 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
      781 actccaccgg cggcatggac gagctgtaca agtaagcttt ctaactaaaa aggcctccca
      841 aatcgggggg ccttttttat tgataacaaa acgctcgtct agggttattt cgatgaaagg
      901 agagagtact aatgccaatt gcaataggaa ataaacggtt acccgttacg ttggatgaaa
      961 agagacaaaa agaattgcag caactaaagc agaagtacgg caaaagtgaa tccaggatta
     1021 tgtgtattgc gttagattta ttgattgccc aagaaaaagc aggatttgag gtaccagcac
     1081 tcaaaaagtg acgtcacttt ttatcctaaa aactaaaagt gatagcactt ttaattataa
     1141 gaagttagaa tattaatcat ttgcttaatt gtacaatata atgtacaatt gttttataga
     1201 aataaataag gggtgaaagg aatggaagca gtagcttatt caaatttccg ccaaaattta
     1261 cgtagttata tgaaacaagt taatgaggat gctgaaacac ttattgtaac aagtaaagat
     1321 gtagaagata cagttgttgt attatcaaaa agagattatg attctatgca agaaacgttg
     1381 agaacacttt ctaataatta cgtcatggaa aaaattcgtc gaggagatga acaattctcc
     1441 aaaggtgcat ttaaaacaca tgacttaatc gaggttgaat ctgatgatta aggcttggtc
     1501 tgatgatgct tgggatgatt atctttattg gcatgagcaa ggaaacaaaa gcaatataaa
     1561 aaagattaac aagttaataa aagatatcga tcgttccccc tttgctggat taggaaaacc
     1621 tgagccatta aagcatgatt tatctggaaa atggtccaga agaattacag atgaacatag
     1681 actgatatat agagttgaaa atgaaacgat atttatttat tctgcaaaag atcactatta
     1741 accaatcgga agtaaggaaa gggtcagaaa cttaaaagtt tttgatcctt attttattta
     1801 ccctagtcat ttaaaaagct aatatagctt agtgttgatt gttattaatg aatgtgtttg
     1861 ttacgcgtat tacggatata aggttagtaa aatcatttct aaagttgagg aaaagtaaat
     1921 ataaatggct taaatttcaa caatttgaag ttgaatagat atgttataat actattgtag
     1981 tgtgggatgt tagttactaa aggatgacgc ttatatatat gactgaatag aataagcaat
     2041 aggtttaata atctatttta aattttttgt actagtttta gttaattacg gttgacagct
     2101 agctcagtcc taggtattgt gctagcctag tatcgatctc cataactatc ctatagatca
     2161 tgtcccgcct gctgctgtgt atctgtgttc tgctgttctc ttctgtggcg tggtctaaac
     2221 cgcagcagtt ttatgtgggc gtactggcta actggggtca tcagcaagcc gttgaacgtt
     2281 ggaccccgat gatggagtat ctgaacgaac acgtgccgga cgcggaattt cacgtctacc
     2341 cgggcaactt caaagcactg aacctggcaa tggaactggg ccagattcag ttcattatca
     2401 ctaacccggg ccaatatctg tacctgagca atcagtaccc gctgtcttgg ctggcgacca
     2461 tgcgttctaa gcgtcacgat ggtaccactt ctgcgatcgg ttccgccatt attgtccgcg
     2521 cggacagcga ctaccgcacc ctgtacgacc tgaaaggtaa agtggtggct gcgtccgacc
     2581 cgcatgctct gggtggctac caagcgaccg tcggtctgat gcattccctg ggcatggatc
     2641 cggacacctt cttcggtgaa accaagtttc tgggctttcc actggatccg ctgctgtacc
     2701 aagttcgtga tggcaacgtt gacgcggcca ttaccccact gtgcactctg gaggacatgg
     2761 ttgcacgcgg cgtactgaaa tcttccgatt ttcgtgtgct gaaccctagc cgcccggatg
     2821 gtgtagaatg ccagtgctct accaccctgt acccgaactg gtctttcgct gcgactgagt
     2881 ctgtatccac cgaactgtct aaagaaatca cgcaggcact gctggaactg ccatccgaca
     2941 gcccggcagc tatcaaagcg caactgaccg gctggaccag cccgatctcc caactggcgg
     3001 taatcaaact gttccaagag ctgcacgtaa aaaccccgga ctctagccgt tgggaagccg
     3061 ttaagaagtg gctggaagaa aaccgtcact ggggtatcct gtctgttctg gtgttcatca
     3121 ttgcaacgct gtatcacctg tggattgaat accgcttcca ccaaaaaagc tcctctctga
     3181 tcgaatctga acgtcagctg aaacagcatg ctgttgccct ggaacgtctg caatctgcta
     3241 gcatcgttgg tgaaattggt gcgggtctgg cccacgagat taatcagccg atcgctgcaa
     3301 ttacctctta ttctgaaggt ggcatcatgc gcctgcaagg taaagaacag gcggatacgg
     3361 atagctgcat cgaactgctg gaaaaaatcc acaaacagag cactcgcgca ggcgaagtgg
     3421 tgcaccgcat ccgtggtctg ctgaaacgtc gtgaagcggt gatggtagat gttaacatcc
     3481 tgaccctggt ggaagaatcc atcagcctgc tgcgtctgga gctggcacgt cgcgaaatcc
     3541 agatcaacac tcagatcaaa ggtgaaccgt tcttcattac tgccgaccgc gttggcctgc
     3601 tgcaagttct gattaacctg atcaaaaact ccctggacgc gatcgctgaa tctgataatg
     3661 cccgttctgg taaaatcaac atcgaactgg actttaaaga gtaccaggta aacgtctcca
     3721 tcatcgataa cggtccgggc ctggcgatgg attctgacac tctgatggct acgttttaca
     3781 ctaccaaaat ggatggcctg ggcctgggtc tggcagtctg ccgcgaagtt atcagcaacc
     3841 acgacggcca catcctgctg tccaaccgtg acgacggcgt tctgggctgt gtggcaaccc
     3901 tgaatctgaa aaaacgcggt tctgaagtgc cgatcgaagt ctaataatct agaccaggca
     3961 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     4021 ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta
     4081 taggtacccg gggatcctct agcgtctaag aaaccattat tatcatgaca ttaacctata
     4141 aaaataggcg tatcacgagg ccctttcgtc ttcacctcga gtttacggct agctcagtcc
     4201 taggtactat gctagcagaa atataaagaa cgatctattt atccgcgtac atgcagcagc
     4261 aaatcaacgg cccggtctac ctggtggatg atgatgaagc cattatcgac tccatcgatt
     4321 ttttgatgga gggctacggt tacaaactga actcgtttaa ctgcggcgat cgctttttgg
     4381 cagaagtcga tctcacccag gcaggatgtg taattctgga tgcgcgtatg ccaggcttaa
     4441 ctggtcctca ggtgcaacag ctgctgagcg acgcgaaaag cccgcttgcg gtcatcttcc
     4501 tgaccggcca tggcgatgtt ccgatggcgg ttgatgcgtt caaaaatggc gcgttcgatt
     4561 tctttcaaaa acctgtgccg ggtagcttgc tcagtcagtc aattgccaaa ggcttgactt
     4621 attcaatcga tcaacatctg aaacgtacta accaagcgtt aatcgacacg ctctcggaac
     4681 gcgaagctca aatttttcaa ctggtgattg caggcaacac caacaaacag atggctaacg
     4741 agctttgcgt ggctattcgt accattgagg ttcaccgtag caaactgatg accaaactgg
     4801 gtgttaacaa cctggctgaa ctggttaaac tggcgccgct gctggcacat aaatccgaat
     4861 aataatctag accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     4921 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     4981 tgggcctttc tgcgtttata gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc
     5041 cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg
     5101 agctggttag ctagtcattc aagcattatt atgctgtttt ttgaagtgaa tgtgcggcca
     5161 tctagccgca cattttgcat ctaaaacatg cagtcatcag caaaataata aacttttccc
     5221 caatatgtgg tttaccacaa tttacaggaa ttcactcctg tggtggtgca aatttgaact
     5281 gtgaattgct tcacaaacgc cgctatcgca atgtcagtat gtggtttacc acaatatcta
     5341 atatcactct gctcaataac aatgatgaaa accttaggaa gaagttaatt gtgttaaaca
     5401 gttaactagg ggctttatct aacgctctcc taaggacaac tgtcattggg agatttaaca
     5461 cgcactgacc accatgaagg tgacgctcat gacttacttg ctgaatctca ggagtttatg
     5521 accttttttt tttagatcct atcggtatga gagccctggt agtcatccgc ctgtcccgcg
     5581 tcaccgatgc tacgacttca ccggagcgtc agctggagtc ttgccagcag ctctgcgccc
     5641 agcgcggctg ggacgtcgtc ggggtagcgg aggatctgga cgtctccggg gcggtcgatc
     5701 cgttcgaccg gaagcgcaga ccgaacctgg cccggtggct agcgttcgag gagcaaccgt
     5761 ttgacgtgat cgtggcgtac cgggtagatc ggttgacccg atcgatccgg catcttcagc
     5821 agctggtcca ctgggccgag gaccacaaga agctggtcgt ctccgcgacc gaagcgcact
     5881 tcgatacgac gacgccgttt gcggcggtcg tcatcgcgct tatgggaacg gtggcgcaga
     5941 tggaattaga agcgatcaaa gagcggaacc gttcggctgc gcatttcaat atccgcgccg
     6001 ggaaataccg aggatccctg ccgccgtggg gatacctgcc tacgcgcgtg gacggggagt
     6061 ggcggctggt gccggaccct gtgcagcgag agcgcatcct cgaggtgtat caccgcgtcg
     6121 tcgacaacca cgagccgctg catctggtgg cccacgacct gaaccggcgt ggtgtcctgt
     6181 cgccgaagga ctacttcgcg cagctgcaag gccgcgagcc gcagggccgg gagtggtcgg
     6241 ctaccgcgct gaagcgatcg atgatctccg aggcgatgct cgggtacgcg actctgaacg
     6301 gtaagaccgt ccgagacgac gacggagccc cgctggtgcg ggctgagccg atcctgaccc
     6361 gtgagcagct ggaggcgctg cgcgccgagc tcgtgaagac ctcccgggcg aagcccgcgg
     6421 tgtctacccc gtcgctgctg ctgcgggtgt tgttctgcgc ggtgtgcggg gagcccgcgt
     6481 acaagttcgc cgggggagga cgtaagcacc cgcgctaccg ctgccgctcg atggggttcc
     6541 cgaagcactg cgggaacggc acggtggcga tggccgagtg ggacgcgttc tgcgaggagc
     6601 aggtactgga tctgctcggg gacgcggagc gtctggagaa agtctgggta gcgggctcgg
     6661 actccgcggt cgaactcgcg gaggtgaacg cggagctggt ggacctgacg tcgctgatcg
     6721 gctccccggc ctaccgggcg ggctctccgc agcgagaagc actggatgcc cgtattgcgg
     6781 cgctggccgc gcggcaagag gagctggagg gcctggaggc tcgcccgtct ggctgggagt
     6841 ggcgcgagac cgggcagcgg ttcggggact ggtggcggga gcaggacacc gcggcaaaga
     6901 acacctggct tcggtcgatg aacgttcggc tgacgttcga cgtccgcggc gggctgactc
     6961 gcacgatcga cttcggggat cttcaggagt acgagcagca tctcaggctc ggcagcgtgg
     7021 tcgaacggct acacaccggg atgtcgggtt cagcagcaaa cgacgaaaac tacgctgcac
     7081 accgatgatg cttaagtcga acagaaagta atcgtatgcg aaacgatcct catcctgtct
     7141 cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
     7201 ttactttgca gggcttccca accttaccag agggcggccc aactggcaat tccgacgtct
     7261 gtgcaagtac tactgttctg cagtcacttg cctaagatac tagctccggc aaaaaaacgg
     7321 gcaaggtgtc accaccctgc cctttttctt taaaaccgaa aagattactt cgcgtttgcc
     7381 acctgacgtc taagaaaagg aatattcagc aatttgcccg tgccgaagaa aggcccaccc
     7441 gtgaaggtga gccagtgagt tgattgctac gtaattagtt agttagccct tagtgactcg
     7501 aatttcggcc ggcttgtcga cgacggcggt ctccgtcgtc aggatcatcc gggcaagctt
     7561 tccacacatt ataggtacaa aaagatgcga aagcaaataa attttttggg tttgtaccgt
     7621 acaccactga gaccgcggtg gttgaccaga caaaccacga agctgtcacc ggatgtgctt
     7681 tccggtctga tgagtccgtg aggacgaaac agcctctaca aataattttg tttaagggcc
     7741 caagttcacg agaacgatat agaaagttca aagaggagaa atactagatg cgtaaaggcg
     7801 aagagctgtt cactggtgtc gtccctattc tggtggaact ggatggtgat gtcaacggtc
     7861 ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa ctgacgctga
     7921 agttcatctg tactactggt aaactgccgg taccttggcc gactctggta acgacgctga
     7981 cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat gacttcttca
     8041 agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag gatgacggca
     8101 cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac cgcattgagc
     8161 tgaaaggcat tgactttaaa gaagacggca atatcctggg ccataagctg gaatacaatt
     8221 ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt
     8281 ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac taccagcaaa
     8341 acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg agcacgcaaa
     8401 gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg gagttcgtaa
     8461 ccgcagcggg catcacgcat ggtatggatg aactgtacaa atgatgagag agaagatttt
     8521 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     8581 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     8641 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     8701 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     8761 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     8821 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     8881 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
     8941 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     9001 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     9061 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     9121 ttgggtgcag caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     9181 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     9241 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     9301 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     9361 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     9421 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc
     9481 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     9541 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     9601 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     9661 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     9721 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     9781 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     9841 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     9901 aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     9961 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
    10021 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
    10081 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
    10141 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
    10201 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
    10261 accagtcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
    10321 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
    10381 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac
    10441 caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
    10501 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
    10561 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
    10621 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
    10681 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
    10741 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
    10801 aatcgtcgtg gtattcacac cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
    10861 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga
    10921 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
    10981 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
    11041 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
    11101 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
    11161 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
    11221 aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg
    11281 tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt
    11341 attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt
    11401 ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac
    11461 ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg
    11521 cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa
    11581 ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac
    11641 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg
    11701 agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag
    11761 ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag
    11821 tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc
    11881 gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt
    11941 ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg
    12001 tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga
    12061 gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta attgatttag
    12121 aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg
    12181 actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg
    12241 aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa
    12301 aacgatctca a
//
