LOCUS       sequence-403465-        8665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 empty backbone for strain-specific barcode DNA tag through 
            Tn7 bacterial insertion, Tetracyline selectable marker and 
            far-red-FP fluorescent marker mPlum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8665)
  AUTHORS   Ordon J, Thouin J, Nakano RT, Ma KW, Zhang P, Huettel B, 
            Garrido-Oter R, Schulze-Lefert P
  TITLE     Chromosomal barcodes for simultaneous tracking of near-isogenic 
            bacterial strains in plant microbiota.
  JOURNAL   Nat Microbiol. 2024 Apr;9(4):1117-1129. doi: 
            10.1038/s41564-024-01619-8. Epub 2024 Mar 19.
  PUBMED    38503974
REFERENCE   2  (bases 1 to 8665)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41564-024-01619-8"; journalName: "Nat Microbiol"; date: 
            "2024-04"; volume: "9"; issue: "4"; pages: "1117-1129"
COMMENT     Sequence Label: pBCC091
FEATURES             Location/Qualifiers
     source          1..8665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      308..402
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      504..590
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    complement(623..670)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        join(689..694,695..711,712..717)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             765..1955
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(1117..1136)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    complement(2035..2082)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        join(2110..2115,2116..2131,2132..2138)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2146..2162
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             2185..2868
                     /codon_start=1
                     /product="far-red monomeric derivative of DsRed fluorescent
                     protein (Wang et al., 2004)"
                     /label=mPlum
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK
                     MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGAA"
     primer_bind     2780..2803
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     terminator      2882..2976
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(3125..3130,3131..3148,3149..3155)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3209..3868
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3275..3294)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4210..4515
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4394..4413
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     mobile_element  4669..4834
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        4877..5322
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(5323..5409,5410..6744)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     terminator      6827..6921
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(7042..7061)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7053..7868
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            8035..8143
                     /note="incP origin of transfer"
     rep_origin      8154..8542
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      complement(8573..8659)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 tgccgaattc ggatccggag ggagtgtggg cggacaataa agtcttaaac tgaacaaaat
      121 agatctaaac tatgacaata aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag
      181 tccagttatg ctgtgaaaaa gcatactgga cttttgttat ggctaaagca aactcttcat
      241 tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg tggcgctgca agaattcaag
      301 cttggaggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
      361 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta gactgcgatg
      421 agtggcaggg cggggcgtaa tttcttaagg cagttattgg tgcccttaaa cgcctggggt
      481 aatgactctc tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      541 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgcccgccg
      601 ctgcaagaat tcaagcttgg aggaagttcc tatactttct agagaatagg aacttcggaa
      661 taggaacttc aatgaagaat tctcatgttt gacagcttat catcgataag ctttaatgcg
      721 gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg
      781 cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc
      841 cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc agtcactatg
      901 gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt ctcggagcac
      961 tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga gccactatcg
     1021 actacgcgat catggcgacc acacccgtcc tgtggatcct ctacgccgga cgcatcgtgg
     1081 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
     1141 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
     1201 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg
     1261 cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata
     1321 agggagagcg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     1381 cgcggggcat gactatcgtc gccgcactta tgactgtttt ctttatcatg caactcgtag
     1441 gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga
     1501 cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg
     1561 tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg
     1621 ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca
     1681 ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca
     1741 ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc
     1801 taacttcgat cattggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat
     1861 ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc
     1921 gtcgcggtgc atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa
     1981 cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg gcttgaagtt
     2041 cctatacttt ctagagaata ggaacttcgg aataggtact tccgctacta gaattcgagc
     2101 ggggaattct tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac
     2161 aattttcaca caggaaacag ctaaatggtg agcaagggcg aggaggtcat caaggagttc
     2221 atgcgcttca aggagcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     2281 gagggcgagg gccgccccta cgagggcacc cagaccgcca ggctgaaggt gaccaagggt
     2341 ggccccctgc ccttcgcctg ggacatcctg tcccctcaga tcatgtacgg ctccaaggcc
     2401 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     2461 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     2521 tccctgcagg acggcgagtt catctacaag gtgaaggtgc gcggcaccaa cttcccctcc
     2581 gacggccccg taatgcagaa aaagaccatg ggctgggagg cctcctccga gcggatgtac
     2641 cccgaggacg gcgccctgaa gggcgagatg aagatgaggc tgaggctgaa ggacggcggc
     2701 cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca gctgcccggc
     2761 gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta caccatcgtg
     2821 gaacagtacg agcgcgccga gggccgccac tccaccggcg ccgcttaatt agctgagctt
     2881 ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg
     2941 ctcggttgcc gccgggcgtt ttttattggt gagaatctcc cgctttacga attcccatgg
     3001 ggagcatgcg gagttacgtc atccccacct tccgctgcgt tcttcatcga tgcccgctta
     3061 gtgatatgct taaaactcca agcacagcct tctccccggg gcggccgcat taggcacccc
     3121 aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc
     3181 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     3241 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     3301 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     3361 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     3421 ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     3481 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     3541 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     3601 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     3661 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     3721 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     3781 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     3841 ctgcgatgag tggcagggcg gggcgtaaac gcgtggatcc ggcttactaa aagccagata
     3901 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
     3961 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
     4021 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     4081 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     4141 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     4201 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     4261 gtggatgtac agagtgatat tattgacacg cccggacgac ggatggtgat ccccctggcc
     4321 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     4381 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtatc cgttatcggg
     4441 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     4501 ttctggggaa tataaatgtc aggctccctt atacacagcc cccggggatg ctggttgtca
     4561 ttgcccctcc ttcctctaag tgataaggtt acttcctcta aatgaccaag cagggtatct
     4621 aatcctgtta acgctcgctc agagaattcg catgcggagt cgagggagaa ccagataagt
     4681 gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca
     4741 cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac
     4801 ccctcccagt tcccaactat tttgtccgcc cacacgctga gcgaattcca cgcttgtgga
     4861 attcctcgag ggagttcaca tatacctgcc gttcactatt atttagtgaa atgagatatt
     4921 atgatatttt ctgaattgtg attaaaaagg caactttatg cccatgcaac agaaactata
     4981 aaaaatacag agaatgaaaa gaaacagata gattttttag ttctttaggc ccgtagtctg
     5041 caaatccttt tatgattttc tatcaaacaa aagaggaaaa tagaccagtt gcaatccaaa
     5101 cgagagtcta atagaatgag gtcgaaaagt aaatcgcgcg ggtttgttac tgataaagca
     5161 ggcaagacct aaaatgtgta aagggcaaag tgtatacttt ggcgtcaccc cttacatatt
     5221 ttaggtcttt ttttattgtg cgtaactaac ttgccatctt caaacaggag ggctggaaga
     5281 agcagaccgc taacacagta cataaaaaag gagacatgaa cgatgaacat caaaaagttt
     5341 gcaaaacaag caacagtatt aacctttact accgcactgc tggcaggagg cgcaactcaa
     5401 gcgtttgcga aagaaacgaa ccaaaagcca tataaggaaa catacggcat ttcccatatt
     5461 acacgccatg atatgctgca aatccctgaa cagcaaaaaa atgaaaaata tcaagttcct
     5521 gaattcgatt cgtccacaat taaaaatatc tcttctgcaa aaggcctgga cgtttgggac
     5581 agctggccat tacaaaacgc tgacggcact gtcgcaaact atcacggcta ccacatcgtc
     5641 tttgcattag ccggagatcc taaaaatgcg gatgacacat cgatttacat gttctatcaa
     5701 aaagtcggcg aaacttctat tgacagctgg aaaaacgctg gccgcgtctt taaagacagc
     5761 gacaaattcg atgcaaatga ttctatccta aaagaccaaa cacaagaatg gtcaggttca
     5821 gccacattta catctgacgg aaaaatccgt ttattctaca ctgatttctc cggtaaacat
     5881 tacggcaaac aaacactgac aactgcacaa gttaacgtat cagcatcaga cagctctttg
     5941 aacatcaacg gtgtagagga ttataaatca atctttgacg gtgacggaaa aacgtatcaa
     6001 aatgtacagc agttcatcga tgaaggcaac tacagctcag gcgacaacca tacgctgaga
     6061 gatcctcact acgtagaaga taaaggccac aaatacttag tatttgaagc aaacactgga
     6121 actgaagatg gctaccaagg cgaagaatct ttatttaaca aagcatacta tggcaaaagc
     6181 acatcattct tccgtcaaga aagtcaaaaa cttctgcaaa gcgataaaaa acgcacggct
     6241 gagttagcaa acggcgctct cggtatgatt gagctaaacg atgattacac actgaaaaaa
     6301 gtgatgaaac cgctgattgc atctaacaca gtaacagatg aaattgaacg cgcgaacgtc
     6361 tttaaaatga acggcaaatg gtacctgttc actgactccc gcggatcaaa aatgacgatt
     6421 gacggcatta cgtctaacga tatttacatg cttggttatg tttctaattc tttaactggc
     6481 ccatacaagc cgctgaacaa aactggcctt gtgttaaaaa tggatcttga tcctaacgat
     6541 gtaaccttta cttactcaca cttcgctgta cctcaagcga aaggaaacaa tgtcgtgatt
     6601 acaagctata tgacaaacag aggattctac gcagacaaac aatcaacgtt tgcgccaagc
     6661 ttcctgctga acatcaaagg caagaaaaca tctgttgtca aagacagcat ccttgaacaa
     6721 ggacaattaa cagttaacaa ataaattcgc tgagcgagga tgcacatgtg accgagggaa
     6781 gcttgcggcc gcgtcgtgac tgggaaaacc ctggcgacta gtcttggact cctgttgata
     6841 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     6901 gcgtttttta ttggtgagaa tccaggggtc cccaataatt acgatttaaa tttgtgtctc
     6961 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     7021 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcagcgt gaaacgagct
     7081 gtagccgtcc gcgtctgaac agcaacatgg atgcggatct gtatggctat aaatgggcgc
     7141 gtgataacgt gggtcagagc ggcgcgacca tttatcgtct gtatggcaaa ccggatgcgc
     7201 cggaactgtt tctgaaacat ggcaaaggca gcgtggcgaa cgatgtgacc gatgaaatgg
     7261 tgcgtctgaa ctggctgacc gaatttatgc cgctgccgac cattaaacat tttattcgca
     7321 ccccggatga tgcgtggctg ctgaccaccg cgattccggg caaaaccgcg tttcaggtgc
     7381 tggaagaata tccggatagc ggcgaaaaca ttgtggatgc gctggccgtg tttctgcgtc
     7441 gtctgcatag cattccggtg tgcaactgcc cgtttaacag cgatcgtgtg tttcgtctgg
     7501 cccaggcgca gagccgtatg aacaacggcc tggtggatgc gagcgatttt gatgatgaac
     7561 gtaacggctg gccggtggaa caggtgtgga aagaaatgca taaactgctg ccgtttagcc
     7621 cggatagcgt ggtgacccac ggcgatttta gcctggataa cctgattttc gatgaaggca
     7681 aactgattgg ctgcattgat gtgggccgtg tgggcattgc ggatcgttat caggatctgg
     7741 ccattctgtg gaactgcctg ggcgaattta gcccgagcct gcaaaaacgt ctgtttcaga
     7801 aatatggcat tgataatccg gatatgaaca aactgcaatt tcatctgatg ctggatgaat
     7861 ttttctaata attaattgga ccgcggtccg cgcgttgtcc ttttccgctg cataaccctg
     7921 cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg
     7981 attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag
     8041 gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca
     8101 cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccgtaggccg gccgatctga
     8161 agatcagcag ttcaacctgt tgatagtacg tactaagctc tcatgtttca cgtactaagc
     8221 tctcatgttt aacgtactaa gctctcatgt ttaacgaact aaaccctcat ggctaacgta
     8281 ctaagctctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
     8341 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
     8401 agaaaaaaaa gaatatataa ggcttttaaa gcctttaagg tttaacggtt gtggacaaca
     8461 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca
     8521 caggaacact taacggctga catggggcgc gcccagctgt ctagggcggc ggatttgtcc
     8581 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
     8641 gagcctttcg ttttatttga tgcct
//
