LOCUS       sequence-467488-       20892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His-tagged Eggerthella lenta DSM 2243 El Vadh with a 
            cumate inducible promoter in Gordonibacter urolithinfaciens DSM 
            27213 for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 20892)
  AUTHORS   Dong X, Bae M, Le C, Aguilar Ramos MA, Balskus EP
  TITLE     Enantiocomplementary Gut Bacterial Enzymes Metabolize Dietary 
            Polyphenols.
  JOURNAL   J Am Chem Soc. 2025 Mar 5;147(9):7231-7244. doi: 
            10.1021/jacs.4c09892. Epub 2025 Feb 24.
  PUBMED    39993729
REFERENCE   2  (bases 1 to 20892)
  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: 
            "10.1021/jacs.4c09892"; journalName: "J Am Chem Soc"; date: 
            "2025-03-5- 5"; volume: "147"; issue: "9"; pages: "7231-7244"
COMMENT     Sequence Label: pXD80ENSF14-Pct3-ElVadh(cHis6)-Accs(ElDSM2243)
FEATURES             Location/Qualifiers
     source          1..20892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(19..813)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     1300..1318
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    1475..1494
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             complement(1497..2108)
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    2512..2539
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     CDS             5911..5928
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      14516..14543
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      14675..14702
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(15134..15985)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(15972..16811)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(17322..18293)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELSERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            19532..19619
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      19961..20355
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     terminator      complement(20518..20561)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     oriT            20657..20766
                     /note="incP origin of transfer"
ORIGIN
        1 ggcgggctgg ccggccttct aaaacaattc atccagtaaa atataatatt ttattttctc
       61 ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg acatactgtt cttccccgat
      121 atcctccctg atcgaccgga cgcagaaggc aatgtcatac cacttgtccg ccctgccgct
      181 tctcccaaga tcaataaagc cacttacttt gccatctttc acaaagatgt tgctgtctcc
      241 caggtcgccg tgggaaaaga caagttcctc ttcgggcttt tccgtcttta aaaaatcata
      301 cagctcgcgc ggatctttaa atggagtgtc ttcttcccag ttttcgcaat ccacatcggc
      361 cagatcgtta ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc tattcgtata
      421 gggacaatcc gatatgtcga tggagtgaaa gagcctgatg cactccgcat acagctcgat
      481 aatcttttca gggctttgtt catcttcata ctcttccgag caaaggacgc catcggcctc
      541 actcatgagc agattgctcc agccatcatg ccgttcaaag tgcaggacct ttggaacagg
      601 cagctttcct tccagccata gcatcatgtc cttttcccgt tccacatcat aggtggtccc
      661 tttataccgg ctgtccgtca tttttaaata taggttttca ttttctccca ccagcttata
      721 taccttagca ggagacattc cttccgtatc ttttacgcag cggtattttt cgatcagttt
      781 tttcaattcc ggtgatattc tcattttagc catgatgttt aactttgttt tagggcgact
      841 gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
      901 cgcttgctgc ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc
      961 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     1021 gcgtgagcgc atacgctact tgcattacag cttacgaacc gaacaggctt atgtccactg
     1081 ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga
     1141 agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcattc
     1201 gaaaccggta tcccgccgcc gccgccggat accggtttcg aattgagatt gccacctgac
     1261 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     1321 tttcgtcttc aaggcgaaaa aaccccgccg aagcggggtt ttttgcgaaa tataatgacc
     1381 ctcttgataa cccaagaggg cattttttag gaacggttat ttgattatta atccggcggc
     1441 aactgaaacg attcggatcc tgtattacta ttctgaattc gcggccgctt ctagatctag
     1501 cgcttgaatt tcgcgtaccg ctctcgcgca atttcgagtg tcgagttcct gacacgctca
     1561 aagcgttctt tgtccttctg ccataggcta cgaacagcaa gcccacgcac cgaattgaat
     1621 atcaaccaaa ggatatcttc tgcatcatca cgcgaaagac cacggctcac cagaacacca
     1681 agccacatat cctcgacgac aaaccgattc ctctctaccg tgcgctgaat accctcgcgt
     1741 aacgctgggt cccggtcggc agccacaatc aaatcaaggc tgatagagaa gtcatcgtcg
     1801 aggaaaaatt cggcggcgtc gtccagcatt tgctggatga cgtcatcctc tggcttcaat
     1861 ttcgctaatc gagcccgact gcgttcggtg atctgttcgt aaagccattc aaaagtggca
     1921 agcagaagct caagctttgt cgggaaatga tggctctgcg ctcctctgga gacaccagca
     1981 gcaccgggca catctgcgat ccggaatccc gcgtaacctt tttcccgtaa aacccccagg
     2041 gccgctgcaa tcaacttgcc ctgggtctcc attgcgcgct ctgcctgtgt tcttctcttt
     2101 ggactcatga tcaccagcga cctgctttcg ttcgtccggg catcacccgg gttactatct
     2161 agtatcaata actagattat tggttgaaac gtactatgtc aagcagttct aaagattgac
     2221 tccaaacggt cgccttgcta tgctcgtttc gtttcctccg cacgcgcgcg aaaggctcct
     2281 tcacggagcc aatgtacgca aaaaaccccg cttcagcggg gttttttcgc caaaaaaaac
     2341 cccgccctgt caggggcggg gttttttttt ttagggccct actgactgtg agcccgctcg
     2401 atcaacggat cggtaaaata cgggcgaaaa agagaatccc gagctttttt gttgccgcat
     2461 agcgtatagt aagcatgtgc cctcgaggtg agggcgagtg ttgcggccag taacaaacag
     2521 acaatctggt ctgtttgtat tatgcagcac acagtgttac ctcctcgggg acgaggagag
     2581 gaaagaggat ttaaaatgac tcagctcacg ctcacgcgcc gcagcttcat gaaggcgatg
     2641 gcggtcaccg gtgccgctgc gtcgctcacg gcggtcgccg agccgatgcg ggcgcttgcc
     2701 gagggcgtcg atgccgatgc gggggaagtc aagcgcgttc gctcgtgctg ccgcgcttgc
     2761 ggcaaggtgg agtgcggcgt ttgggtaacc gtgcgcgacg gcaaggtcgt caaggtggaa
     2821 ggcgacgagt ccgcgccgca aagccgcggg cactgctgct cgaagtcgca gtcatcgatg
     2881 caggcgctat atcatcccga ccgccttcgt ttcccggtga agcgcacgaa ccctaagggc
     2941 gaggacgatc ccggttgggt gcgcatcacg ctggacgagg cgttcgagat atgcggcgag
     3001 aagctcaagg aagtgaagga gaagtacggc ggggagtcca tcttcgtgat gtgcggcacg
     3061 tcgcgcgtgt ggtcgctcgg gccttaccag ggcatgaagc agctgttcgg cacgccgaac
     3121 gcgcatttgg cctaccaggt gtgcaaaggc ccgcgccact tcggcggcat catgaccgac
     3181 gagatggggt ccccgtggat ggaggtggag gccgagccca gcgtgtacgt gcagtggggc
     3241 acggcgtgcg agtactccaa ctacgactcc acgaaccgca cggtcaccga cgtcgcgcat
     3301 cgtgcgtcta agcatatcgt ggtggatccg cgcgtcacgc ctttgggcaa ggaagccgac
     3361 atctggctgc cgcttcgtcc cggcaccgac ggcgcgctgg cgctgtcgtg gctcaactgg
     3421 atcatcgaaa acgaagctta cgatgacacg atggtgcgtc gctggtcgaa cgcgtcgttc
     3481 ctctacgtgg acgacaagcc cgagctgacg caaggctggt tggtggaggg caacggcggc
     3541 atcaacatga agacgaagct gctgaccgag gccgacctca aggaagatgg caagtaccag
     3601 cgcttcatgg tgtgggacga ggcgaacgag cgtctcacgt actgggatgc cgagcttggc
     3661 atgtgggagg gcgaggagca ccgcatcccc accaccggca catggatcga gcatccgtac
     3721 aagccgcttg ttgccgatgc atggctgccc gatcagtcga cgttcgccga tccggctacc
     3781 gagcccgacc gcttccccga cggtttcgag gaatgcaatc ccaaggggtt gccgaagcgc
     3841 ccatctctgc ttcctggtga ggtagaggta acgctcaaag acggaaccgt gcacaaggct
     3901 cgctctgtgt gggatgcgtt ccatcagatg accagcgagt acaccatgga gaaggcagag
     3961 gagatcaccg gcgtgccggc ggcgaagagc gaggaggctg tacgcgcctg gacgacgcgc
     4021 gtcaatccgc tgcatggcaa cggcggcatc cacttccagc tggccaccga ccagaacggc
     4081 aactccatcc agaacgtgcg tgcgctgcag atcctgtcgt gcatcaccgg caactccgac
     4141 gagccggccg gcaaccgcgg ctcgtcgaag gcgcagttcg acggcaatcc agggcgttcg
     4201 aacatgcagg ccggcgcgcc gtacggcgac gaggccaaga tgtgggacgg ccgcgacgtc
     4261 gacctcgaag ggctcgccgc gaagatgcag gatttcgtgc agtacctcat cgacaacgat
     4321 tcgcccctgg ccgagcgcta cggcaacaag gtgcccagcc atgaggaagc cctcgttatc
     4381 gccaagcgca tgggcggcgc attccgcacg tcgcaggcgt ggccgaaccc caagaccgtg
     4441 ttcgagcgac aagccaacga ggttgacgcc gagcgattcc ccctcaaccg ctactgggcg
     4501 cgctgggcgg acgcgaactc catctgggac ggctgcctgg acaacgacgt gccgtaccag
     4561 ttgcatgcgg gcgtgtgcca gtcgggcgac ttcatgaata tgggcaacat cgacgtggcg
     4621 tgggaggcca tgacgaagct ggacttcttc accgacatca acctgtggtt ctgcccgaac
     4681 aacggcaacg ccgacgtcat ctttccgtgc taccactggt tggaagttga cacgacgcgc
     4741 gtcagccagg gcgccggcgg gttcttcggc tgcgggtgcg ccgctgtcga ggcgccgggc
     4801 gagtgcatct acgatcccga ctggaacgtg ggcatgtaca aggccatggg cgtgccatgg
     4861 aacacgaagg acgagtcggc cgagccgtcc gagatcctca acttcggcga gaagaccgac
     4921 taccgttggc cgaaccgcag ccgcgtgctc aaggacaacg tggacgcgtg gaagacagcc
     4981 gagttccccg acggtcccac gtgggagcaa gcgaaggagc acttccagaa gaacggctgg
     5041 atggattgcc gcacgtggca tcccgagcgt tggggcacgt atcgccggca cgagatgggc
     5101 tggcgtcgcc agcagggcgg cttcaacctg ttcccgttgg tggacgacca tccgggcttc
     5161 atgacgccgt cgggcctcat tgaaatctgg tcgcttgtgt gcgaggcgta cttaggcgac
     5221 gaggacaagt tccccgtgta ccgcgaaccc acgaactcgc cggtgaccac gcccgagtac
     5281 ttcgacgccg cgaaggttgc cgacatcgat gagtcgaagc tgcgcaatat ggattatccc
     5341 gccagcctgc aggagcacgc cgatgcgacg ttcctcatga ccacgggcgc gcgccagccg
     5401 gtgtacttcc acagcgagca tcgccagctg ccgtggtgcc gcgagctgtg gcccgcgccg
     5461 cgcctggaga tgaacccgaa cgacgctgcg cgtctggggc tcgagcaggg cgactgggtg
     5521 tggatcgaga gcccgtgggg caaggtgcgc gaggtgctgg atctgtacta cggcatcagc
     5581 aagggcgtgg tgaacgcgaa ccacgcgtgg tggttccccg agatggacac ggccagccac
     5641 ggctacgagc tggtgaacat caactgcgtg atggatcctt acggccagga tgtggtgtgc
     5701 ggtgcagcaa cgatgcgctc ggttcccgtg ctggtttata aggctacggc ggaaaactcg
     5761 ccgttcggca acccggtgcc gtgcgatccg cagggtaacc cgtgcatctg cgatgcgggc
     5821 gatccgcggt tgaaggaatg gatgggaacg ggattgcgat ctcgcgttga aggcgaagag
     5881 gcgtggaccg gagagggggc gggcagcagc catcaccatc accatcacta gacgatgaca
     5941 cagtacgcaa tcatcaccga tttgaaccgg tgcgtcggat gccttgcgtg cacggtcgca
     6001 tgcaaagcgg tgaacaacgt gcccgtcggg gcgttttgga tcaagacgct gcgcgtgggt
     6061 ccgaatccga tcgagggcgg aagcggcgat tggcccgacg ttgagatgta tttcctgccc
     6121 atccaatgcc agcattgcgc cgatcccgag tgcgtgaagg tgtgtcccac gggcgcctcg
     6181 cataaggcag ccgacggcac cgtgcagatc gacaagtcga agtgcattgg ctgccagttc
     6241 tgcgcgatga gctgccccta caacgttcgc tacctcaacg aagaggagcg cgtggtggag
     6301 aagtgcacgc tgtgcgagca gaagatcgcc caaggcgagc ttccgcagtg cgtggcgcaa
     6361 tgcggctcgc gtgcgcgctt cttcggcgac ttggagcagg gcgtggacgg gttcgaggca
     6421 ccggctccgc ctcaagccct gtcgtgcgac tacgccgaaa tgcagcagac gcgcgtcacg
     6481 ctgaaggagt acgtcgaacc gtatacggaa gacgagatcc atcgtctgcc cgatgttgga
     6541 aatggcccgg agctcatgta cctgctgcgc tccgaccgaa agtggagggg gtagaccatg
     6601 gaactgcaat ggcctttgat cgtgtttacc accctggttg cctggtcggc ggggctgttc
     6661 gggacgcagg cgctgatggc ggtgttcggc gtcgggaaga aggcgcaggt gccggcttgg
     6721 gtctgctcgg ccgtgctgct ggcggccggc gggatcgcgg tgttcttcca tctcgagcac
     6781 tgggagcgga tattcaacgg gttcgggcac ctgacgagcg ggatcaccca ggaattgatc
     6841 gcgatcatcg tgcttgccgt ggtggctatc gtctacttgg tgctcatgcg caagtccgac
     6901 gacggcgcga gcgtgccgaa gtggctggcc tgggtcgccg tcgcgctgtc ggtggtgctg
     6961 gtcgccgtga tggcgcactc gtacacgatg gccgcccgcc ccgcctggga ctcggccctc
     7021 tggatcctct acgtcctcgg caacgcctgc gtgttgggcc ccgcgacgtt cgccctcctc
     7081 tccgccttgg ctgcaggggg accccgggac cagcccgccg agcgcgcagc cgacgccggg
     7141 gcccccgcgg gtaggacgcc cctggtaggc gcagccgcca acgccctcgc aaccctcgcc
     7201 ttcgccgcct tcctccagct ctccgccggc tccttcgccg acgtcggcct ctacttcgac
     7261 ccgacgcacc ccaccaaggc catggccgac gcagccgcga cggtcgcctc ccaggccccg
     7321 ctcctgtggc tgggcgccgt ggccgtcggc gcgatcgtcc ccctggccgc cgccttccta
     7381 ggcaggcgca cggggaactg gaagctctgg gccccggcag ccataatcgc cgccctcgtc
     7441 ggagccgtct gcatgcgcgt cgtcttctac aacctggggc tctcggtctt catgttctac
     7501 tagactagtc cgctggttgc cacgacgccc tcgaagtctc cttcgagggc gttctgtcgt
     7561 ttgcaagaag ggtctggaga ggcaaatcgc gctcgaaggg gcttcgaggc cggggtaggt
     7621 cgggagggtg gaatccggat gtttcacgtg aaacatccgt tcttttacgg ccggtgtttc
     7681 gcgtgaaaca ttttgcggaa aggactcctt cccttcccgt gcgccacccc aacgccccgc
     7741 tcctagcgct cgtacacctg gtcgcagttg agcagatcct tcatggagtg ccagatctgg
     7801 tgcgggttgc ccggcgggtt cttgagcggc ttcatcggat agcgctcctg cgcgcccgac
     7861 agcagatcga cggcgaacac ttcgtcggac agctcgagcg cgccttccgg gcagatgtcg
     7921 gtgcagcagc ggcacttcac gcaatcgacc ggacgatgcg tcacgccgat cgagccgtcc
     7981 tcacccgcgt acttcgcgat ggccccggtg gggcagaacg tggcgcacat ctggcacgac
     8041 gagcacttct cgggatcgat gatcacatgg ccccacaagc gcgtgtccat gagcacatcc
     8101 tgcggctgcc ccaacgaatc gagcgcggcc agcaaacgtt cgcggcggtc gggaatgaag
     8161 tgaggcagcg tgccgtcctt gcccaccttc acgaacctcg gctcccgccg ctcctccaca
     8221 cccaatgcgt ccttgacggc gtagtcagcc gtcaccgccg caacgttctt agcctcgtcc
     8281 ttcatgctgg agaagaagct ccgacggctc gcgtcgtagc cctcgtcgcc cgcccggcgc
     8341 acggatgagg gaagcttttc cgccaggtcg atgcgcatat cgttgttcca ggttgcaagc
     8401 agcgcgcttg ccgtgtcgca cacaacccgc gccatctcga aacccgagac atgctcgcat
     8461 tgcgcgcagc gcgcctgcac gaggctcacg cggcgcacgc ccaacgccgc cagcgtgacc
     8521 agcaggctct cctccacgcg ccccaggcac gtgacgccca ccaccttctc ggggtcgtac
     8581 aacccgtcgg ccgcggcaag cagctgatcg caggcgataa ccacctgtcc gtcggccacg
     8641 tcagccgccg cgcggcatgc ttgcagaagc tcggcatcgt tcggacgatg cgcttcaagc
     8701 gcgcaggtgg gacacacggt ggcgcaggtt ccgcacccga tgcaccgctc gggttcgatg
     8761 accagctcgt tgtcgtcata ggagatgcaa ccgctcgtgc acacgtcggc gcacttcatg
     8821 cacgttgcgt tgcgattgcg cacaatagcg caacgaccct gatgaaccgt aatgtcggcg
     8881 ctctgaagtt gttcgagcag accgaggaaa ttcgtcacgt gagccatgac ttcacccttt
     8941 ctcgtctacc cccttgcacc gcacagtata accactttcg ccgtctcacg cgagaaaaaa
     9001 cgcgcttgac accttcgaca ggcctcccta tcatgtggcc ttggggagtc cgccggcacg
     9061 ctgatcgcgt gcgaagcgga cattgtttgc aaaagtggga acctcagggg tgaggtttcg
     9121 cggtgctgtg cgccgtaagt gcaaacaatg tctgacgagg ggcggaacat gactcacgat
     9181 tccacgaacc agcatctttc tcttgagccc gactacgacg ggttcgatcc acttgaagat
     9241 ttcgaagacg atgcgaccga ggtcgaaggc cgtgccgacg agttcgtcgc cacggtgagc
     9301 gccgccgaag acgcgctcgc tcacgacgcg atggcacccg ccacgcccgc catctgcgaa
     9361 aagcccgctg aagagcgcat cgccgacttg ttggaggcca tggcgccgcg ccgcacggtg
     9421 ctgctgggca tcctggccgc ctgcgccgag ccgcagccgg catccgccgt gaacgctcgc
     9481 gtcgacgagc tgcaggagaa caacttctcg gtgtacagcg cggccaacct gtgctccttg
     9541 ctggagaagg cgggcgccat cgagcgcgtc acggccgagg gcgaacccgc ggaggacatc
     9601 gaaatcgagc cacagaccgt cgtcgtcgac ggcgtcgaat acctggaagc ccgcgaaccc
     9661 gtcgagatct actggcacat caccgagccg ggccgcgcgg cgcttgaggc cgacaaaccg
     9721 ctcgagcgcc tgcgcgccct cctggacgaa gatgccgcct acgcgcccat ctaccagcgc
     9781 atcctccgct tgtgcacagc agacggcggc gccacgacac ccgccatcaa caacgcggtc
     9841 gaccacaacc cgctggtgca gaagccgcgc ttctacgcgc cccacttcgt cgaccgcctg
     9901 gagaaatgcg acgcgctggc ctggaagaag gcgtggtgca tcaccgacat cggacgcgcc
     9961 ggcctcgata tgctggcaga cgtcgtagac gagaacgcgc ccgccaccca gcccgaaacc
    10021 cccgcaaccc ctgatcccgc cgcatccaag gaggactaac catgaccgaa gcaaccgcgc
    10081 ccgcaaccga gaccctcgcc gacgaagcga ccatgcagga gctcgtcaaa ctgaacgagc
    10141 agcgcgccgc cacgtacggc ctgctgtcgc gcctgtaccg cgtggaggtg gaccagcccc
    10201 ttctcgacga gctgcgcagc atgcgcttcc ccgccaagac gggcaacgag aacgtggacg
    10261 agggctaccg cctgctggcc acgtacctga gcaacacctg ggacaacagc gtcaccgacc
    10321 tcgccatcga ctacgtgcgc gtgttcatcg gacacggcat cgacgcgttc tcggccgcct
    10381 acccgttcga atcggtgtac acctccgaga agcgcctgct catgcaggaa gcacgcgacg
    10441 aggtgctggc catctaccgc agcgcaggcc tcgacaagca ggacacatgg aaagagggcg
    10501 aggatcatat cgcgctcgag ctggaattcg agcagatcat ggccacgcgc acggtcgaag
    10561 cgctgcgcaa aggcgacgaa gacgaggccg tcgcgctgct gaccacgcag aagaactttc
    10621 tcgaagacca tctggcggca tgggctccca tgatgacctc cgacatgaag cgattcgcgc
    10681 aaaccgactt ctaccggggc ttggcgtact tgacggacgg gttcctccaa accgaccaaa
    10741 cgttcctcga cgacgtgctg gccgaagact aactgcacag gatcgaaaga aggagcgatg
    10801 gacaacaagc aggagaaaac ctacgccggc atgacgcgac gcacgctgtg cctgggcgca
    10861 ggcggcgctg cggtcatgct aggtctaggc ggattgaagt acgcgggatc gcaaccgctc
    10921 gtgcgcccgc cgggaggcca ggacgaggaa cggctcatct cggcctgcat ccgctgcgag
    10981 aagtgctacg agatctgccc gcgcgacgtc atcgcgcccg cccacctcga ggacggtgtg
    11041 ctgaacatgc gcacccccac gttcgacttc tcggccaact ggtgcgactg gtgcgccgaa
    11101 gagcacgacg gcaccccgtt gtgcgtgagc gcctgcccca ccgaagcgct caagctgccc
    11161 gacgacgcca cggccgagaa caccatcatc ggcaaagccg ttatcaacac ggactggtgc
    11221 ttggcgtaca agctgatcgg atgccggttt tgctacgacg cctgtcccta cgaggccatg
    11281 gagctcgacg agaacaaccg ccccgtcgtg atcgaggaca agtgcaacgg ctgcggcgcg
    11341 tgcgagagcg tgtgcgtctc gctgcagaac ggctccatct ccgaaggagc caccgcgcgc
    11401 gccatcgtcg tccacccgct tgacgaagtt gaaggataag gtgccctcat gaagaagaac
    11461 tccaagaagc tgcgcaccct cgtcgcactg gccatcgtgg ccgtcgtcgc cgtcggcttc
    11521 gtcacgaacg tcgggatcgg caccatgtcg gcaccgggca tctgggacat ctccatccta
    11581 tgcccgctgg gcgccttggg aacgatgctc gcctccaaga tgatggtgcc gcgcgccgta
    11641 atctcgctga tcgtgatggt gctgctcatc gttctgttcg cccgcgcgtt ctgcggatgg
    11701 atttgccccg tgccgctcgt gcagaagttg cgcggcttgt tcgcgaagaa gcaacccgtc
    11761 gaacgcggcg acgatgccga tcctgccgcg gcgccgctca cgaaagtcga gatcgacgcg
    11821 ctgggcgttt catgctcgaa ggggtcgaag aatggcagcg gatgcgcgtc ctgcgcctcc
    11881 aagcgcggag ccgcgctcga tgctcgccac ttcgtgctgg gcggcgcgct ggcgtccacc
    11941 ttcatctttg gattcccggt gttctgcctc gtctgcccca tcggactcac gttcgccacc
    12001 atcctgctgg tcatcaacct gttctcacac ggcgatgtca cctggtcgct catcgtggtg
    12061 cccgcgctgc ttatcgccga ggtgctgctg ttcaagaagt ggtgccataa gctgtgcccg
    12121 ctttcggcgt tcatgagcct gatcgcgaag gcgaacaaga ccttcgtgcc cacgatcgac
    12181 gatgcgaagt gcctggaaac ggcgaagggc ggcgcgtgcg gggcctgcgg ccgcgtctgc
    12241 gaggagggca tcgacccgcg ccacccgcag ctttcgcagg cggcatggag cgagtgcacc
    12301 aagtgccgcg cctgcgtgga cgcatgccct gcgaacgcga tcaccatgcc gctgctgccg
    12361 cggcgaacgc cgtctaccgc agaacctgtc gccgtgccaa cgaacgccga gaagatcgac
    12421 gcggagaatg cgcgccccgt tccgtaaagc ccctcctgcg cgacccggct cgcagctgca
    12481 gcggtctggt cgcgcatccg gaatcgggcc gagcccgacc ccgggcaaaa ggcggcttgg
    12541 caagggcttt gggggacgag ccgccttttg cccgggagcg acatccccgc tccccctccc
    12601 tgcgaaaaga gggatgccgc ttccggcctt catctgacaa ccacgacgtg ggaggacgaa
    12661 aacccatggc atacgaaacg acaagcgaat cgaccgaggc cctgctcgtc accggaggcg
    12721 tcggcaccgg caaaacgcaa cgtctcgtcg agcgcgctgc cgcgctgctg gccgaaaacg
    12781 ccgccgcggg caacgtgctc gtgctgtgcg cgacgcccca ggccgcacgc ctgttctcgg
    12841 agcggttgac agcccgcgtc ggtgccaagc gcgctgccgg cgcaaccgtc gccaccgcac
    12901 gcgccttagc gctcgacgtg ctggccgacg acggcgcgcg ccgctggtcg gggcgcgagc
    12961 cgcgactact gaccgccttc gaagagacgt tcctcatgga ggacatgaag gtgagcgggc
    13021 tgcggccgaa gcggctgcgc gagatgctca agttcttcta ccgctcgtgg accgagctgg
    13081 cggacgacga tcccgattgg ctgctaccgg gcgaggaggc cgatgtccat acgctgttga
    13141 aagagaacct ggtcttcatg cgcgccgtca tagaacccga agcggcgaac cttgctgcac
    13201 gctacctccg agaacacgac gaagcgcggg cggcgcattc ctacgcgcac gtgctgatgg
    13261 acgactacgc ctgcgcaagc cgtgcctcgc agacgctggc gaatctcgtc gcaacctgct
    13321 cgatcaccgt ggcgggcgac gcggaaacct gcatcgagac atacgattcg tacccctatg
    13381 ccgccggaat cgacgaattc ctcgcaacgc atcccgatgc tgcttgcgag cacctggatg
    13441 ccgacgcgcc cgatgccgtc gaacgggtcg ctctggcaag cgacacgccc gaagccgagt
    13501 tcgaacaggt ggcgcgcacg gtggccgaag ccgtcgccgc aggaacgccc gcacggcaga
    13561 tcgtcgtggc cgtaccgaac agcgtgtggt cgcgcaacat cgcaaccgct ttgcacgcac
    13621 gaggcgtgcc aaccgaagcg ctcgccatct cgcagcccct gcgcggcgac gtgcgggaca
    13681 acgcccgctg cacgcctgcg cgcatcgcca cggccctcaa cctcgtggcg aacccagacg
    13741 acaccgcggc ttggcggtgc tggtgtggat tcggcgacta cctggtgaac agcgcggctt
    13801 tcgccagcct gcggacttgc gcgaaggagc gcagtatcgg cctcgtggac gctttggaga
    13861 tgctaagcga aaacgactgc gagcacgtag tgggcgcgca acgcgtggcg accgcctaca
    13921 aggacggtcg cgcgctcatc gagcaggtac gaggattgcg gggcacggca ctgctggacg
    13981 agctgacgcg catcgtcacc gacggagcgg aatcccaagc gcccgccgtt ctgcgcgagc
    14041 tctgcctcga gcacggcaac gcagccgatg aaaacccgga aagcctttac gatgccgatg
    14101 ctgcggcgat ggcgcggcgc atggccgagc gcctgctcgc gcccacgctg gaggttgccg
    14161 acgcggtagc cgtcgtcccc tacgacttcg cctgcggctt ctcgcccgac ctgctgatcg
    14221 tcgccggctt cgtgaacggt ttcatcccct cccgcgacta tttcgacgcc acggtgaccc
    14281 cgcccgagcg ccaggagaag atgcgcgccg tcgacgaacg acgcgtgcgc gcgctcacag
    14341 gaaaggccgg ccgcgcggtg gcgctgtcgt acttcacgtc caccgacctc gaaagcgccg
    14401 gcaagctgaa gctcaagatc agtcgcatcc gcctcaagaa cggtatccgc acctgcacta
    14461 tagagcccag cctattcctg gagcaaatcg cagcgcagga gtagcgactc acactggctc
    14521 accttcgggt gggcctttct gcgtttatat actagagaga gaatataaaa agccagatta
    14581 ttaatccggc ttttttatta tttaattcct gttataaaaa aaggatcaat tttgaactct
    14641 ctcccaaagt tgatccctta acgatccctg caggagaagg ccatcctgac ggatggcctt
    14701 tttgaagctt tctagaatcg ggggcccccg gggggattct agaacaatgg aagtcctcgt
    14761 ctcgccctcg aattagcccg cttaatgagc gggctttttt tgaattaatt ctcgcgagct
    14821 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
    14881 agcgcgaatt gcaagctggc cgacgcgctg ggctacgtct tgctggcgtt cgggagcaga
    14941 agagcataca tctggaagca aagccaggaa agcggcctat ggagctgtgc ggcagcgctc
    15001 agtaggcaat ttttcaaaat attgttaagc cttttctgag catggtattt ttcatggtat
    15061 taccaattag caggaaaata agccattgaa tataaaagat aaaaatgtct tgtttacaat
    15121 agagtggggg gggtcagcct gccgccttgg gccgggtgat gtcgtacttg cccgccgcga
    15181 actcggttac cgtccagccc agcgcgacca gctccggcaa cgcctcgcgc acccgctggc
    15241 ggcgcttgcg catggtcgaa ccactggcct ctgacggcca gacatagccg cacaaggtat
    15301 ctatggaagc cttgccggtt ttgccggggt cgatccagcc acacagccgc tggtgcagca
    15361 ggcgggcggt ttcgctgtcc agcgcccgca cctcgtccat gctgatgcgc acatgctggc
    15421 cgccacccat gacggcctgc gcgatcaagg ggttcagggc cacgtacagg cgcccgtccg
    15481 cctcgtcgct ggcgtactcc gacagcagcc gaaacccctg ccgcttgcgg ccattctggg
    15541 cgatgatgga taccttccaa aggcgctcga tgcagtcctg tatgtgcttg agcgccccac
    15601 cactatcgac ctctgccccg atttcctttg ccagcgcccg atagctacct ttgaccacca
    15661 tggcatcagc ggtgacggcc tcccacttgg gttccaggaa cagccggagc tgccgtccgc
    15721 cttcggtctt gggttccggg ccaagcacta ggccattagg cccagccatg gccaccagcc
    15781 cttgcaggat gcgcagatca tcagcgccca gcggctccgg gccgctgaac tcgatccgct
    15841 tgccgtcgcc gtagtcatac gtcacgtcca gcttgctgcg cttgcgctcg ccccgcttga
    15901 gggcacggaa caggccgggg gccagacagt gcgccgggtc gtgtcggacg tggctgaggc
    15961 tgtgcttgtt cttaggcttc accacggggc acccccttgc tcttgcgctg cctctccagc
    16021 acggcgggct tgagcacccc gccgtcatgc cgcctgaacc accgatcagc gaacggtgcg
    16081 ccatagttgg ccttgctcac accgaagcgg acgaagaacc ggcgctggtc gtcgtccaca
    16141 ccccattcct cggcctcggc gctggtcatg ctcgacaggt aggactgcca gcggatgtta
    16201 tcgaccagta ccgagctgcc ccggctggcc tgctgctggt cgcctgcgcc catcatggcc
    16261 gcgcccttgc tggcatggtg caggaacacg atagagcacc cggtatcggc ggcgatggcc
    16321 tccatgcgac cgatgacctg ggccatgggg ccgctggcgt tttcttcctc gatgtggaac
    16381 cggcgcagcg tgtccagcac catcaggcgg cggccctcgg cggcgcgctt gaggccgtcg
    16441 aaccactccg gggccatgat gttgggcagg ctgccgatca gcggctggat cagcaggccg
    16501 tcagccacgg cttgccgttc ctcggcgctg aggtgcgccc caagggcgtg caggcggtga
    16561 tgaatggcgg tgggcgggtc ttcggcgggc aggtagatca ccgggccggt gggcagttcg
    16621 cccacctcca gcagatccgg cccgcctgca atctgtgcgg ccagttgcag ggccagcatg
    16681 gatttaccgg caccaccggg cgacaccagc gccccgaccg taccggccac catgttgggc
    16741 aaaacgtagt ccagcggtgg cggcgctgct gcgaacgcct ccagaatatt gataggctta
    16801 tgggtagcca ttgattgcct cctttgcagg cagttggtgg ttaggcgctg gcggggtcac
    16861 tacccccgcc ctgcgccgct ctgagttctt ccaggcactc gcgcagcgcc tcgtattcgt
    16921 cgtcggtcag ccagaacttg cgctgacgca tccctttggc cttcatgcgc tcggcatatc
    16981 gcgcttggcg tacagcgtca gggctggcca gcaggtcgcc ggtctgcttg tccttttggt
    17041 ctttcatatc agtcaccgag aaacttgccg gggccgaaag gcttgtcttc gcggaacaag
    17101 gacaaggtgc agccgtcaag gttaaggctg gccatatcag cgactgaaaa gcggccagcc
    17161 tcggccttgt ttgacgtata accaaagcca ccgggcaacc aatagccctt gtcacttttg
    17221 atcaggtaga ccgaccctga agcgcttttt tcgtattcca taaaaccccc ttctgtgcgt
    17281 gagtactcat agtataacag gcgtgagtac caacgcaagc actacatgct gaaatctggc
    17341 ccgcccctgt ccatgcctcg ctggcggggt gccggtgccc gtgccagctc ggcccgcgca
    17401 agctggacgc tgggcagacc catgaccttg ctgacggtgc gctcgatgta atccgcttcg
    17461 tggccgggct tgcgctctgc cagcgctggg ctggcctcgg ccatggcctt gccgatttcc
    17521 tcggcactgc ggccccggct ggccagcttc tgcgcggcga taaagtcgca cttgctgagg
    17581 tcatcaccga agcgcttgac cagcccggcc atctcgctgc ggtactcgtc cagcgccgtg
    17641 cgccggtggc ggctaagctg ccgctcggac agttcgaggc tggccagcct gcgggccttc
    17701 tcctgctgcc gctgggcctg ctcgatctgc tggccagcct gctgcaccag cgccgggcca
    17761 gcggtggcgg tcttgccctt ggattcacgc agcagcaccc acggctgata accggcgcgg
    17821 gtggtgtgct tgtccttgcg gttggtgaag cccgccaagc ggccatagtg gcggctgtcg
    17881 gcgctggccg ggtcggcgtc gtactcgctg gccagcgtcc gggcaatctg cccccgaagt
    17941 tcaccgcctg cggcgtcggc caccttgacc catgcctgat agttcttcgg gctggtttcc
    18001 actaccaggg caggctcccg gccctcggct ttcatgtcat ccaggtcaaa ctcgctgagg
    18061 tcgtccacca gcaccagacc atgccgctcc tgctcggcgg gcctgatata cacgtcattg
    18121 ccctgggcat tcatccgctt gagccatggc gtgttctgga gcacttcggc ggctgaccat
    18181 tcccggttca tcatctggcc ggtggtggcg tccctgacgc cgatatcgaa gcgctcacag
    18241 cccatggcct tgagctgtcg gcctatggcc tgcaaagtcc tgtcgttctt catcgggcca
    18301 ccaagcgcag ccagatcgag ccgtcctcgg ttgtcagtgg cgtcaggtcg agcaagagca
    18361 acgatgcgat cagcagcacc accgtaggca tcatggaagc cagcatcacg gttagccata
    18421 gcttccagtg ccacccccgc gacgcgctcc gggcgctctg cgcggcgctg ctcacctcgg
    18481 cggctacctc ccgcaactct ttggccagct ccacccatgc cgcccctgtc tggcgctggg
    18541 ctttcagcca ctccgccgcc tgcgcctcgc tggcctgctg ggtctggctc atgacctgcc
    18601 gggcttcgtc ggccagtgtc gccatgctct gggccagcgg ttcgatctgc tccgctaact
    18661 cgttgatgcc tctggatttc ttcactctgt cgattgcgtt catggtctat tgcctcccgg
    18721 tattcctgta agtcgatgat ctgggcgttg gcggtgtcga tgttcagggc cacgtctgcc
    18781 cggtcggtgc ggatgccccg gccttccatc tccaccacgt tcggccccag gtgaacaccg
    18841 ggcaggcgct cgatgccctg cgcctcaagt gttctgtggt caatgcgggc gtcgtggcct
    18901 gcccgctcta atgcccggtt ggcatggtcg gcccatgcct cgcgggtctg ctcaagccat
    18961 gccttgggct tgagcgcttc ggtcttctgt gccccgccct tctccggggt cttgccgttg
    19021 taccgcttga accactgagc ggcgggccgc tcgatgccgt cattgatccg ctcggagatc
    19081 atcaggtggc agtgcgggtt ctcgccgcca ccggcatgga tggccagcgt atacggcagg
    19141 cgctcggcac cggtcaggtg ctgggcgaac tcggacgcca gcgccttctg ctggtcgagg
    19201 gtcagctcga ccggcagggc aaattcgacc tccttgaaca gccgcccatt ggcgcgttca
    19261 tacaggtcgg cagcatccca gtagtcggcg ggccgctcga cgaactccgg catgtgcccg
    19321 gattcggcgt gcaagacttc atccatgtcg cgggcatact tgccttcgcg ctggatgaag
    19381 tcggccttgg ccctggccga ttggccgccc gacctgctgc cggttttcgc cgtaaggtga
    19441 taaatcgcca tgctgcctcg ctgttgcttt tgcttttcgg ctccatgcaa tggccctcgg
    19501 agagcgcacc gcccgaaggg tggccgttag gccagtttct cgaagagaaa ccggtaagtg
    19561 cgccctcccc tacaaagtag ggtcgggatt gccgccgctg tgcctccatg atagcctacg
    19621 agacagcaca ttaacaatgg ggtgtcaaga tggttaaggg gagcaacaag gcggcggatc
    19681 ggctggccaa gctcgaagaa caacgagcgc gaatcaatgc cgaaattcag cgggtgcggg
    19741 caagggaaca gcagcaagag cgcaagaacg aaacaaggcg caaggtgctg gtgggggcca
    19801 tgattttggc caaggtgaac agcagcgagt ggccggagga tcggctcatg gcggcaatgg
    19861 atgcgtacct tgaacgcgac cacgaccgcg ccttgttcgg tctgccgcca cgccagaagg
    19921 atgagccggg ctgaatgatc gaccgagaca ggccctgcgg ggctgcacac gcgcccccac
    19981 ccttcgggta gggggaaagg ccgctaaagc ggctaaaagc gctccagcgt atttctgcgg
    20041 ggtttggtgt ggggtttagc gggctttgcc cgcctttccc cctgccgcgc agcggtgggg
    20101 cggtgtgtag cctagcgcag cgaatagacc agctatccgg cctctggccg ggcatattgg
    20161 gcaagggcag cagcgcccca caagggcgct gataaccgcg cctagtggat tattcttaga
    20221 taatcatgga tggatttttc caacaccccg ccagcccccg cccctgctgg gtttgcaggt
    20281 ttgggggcgt gacagttatt gcaggggttc gtgacagtta ttgcaggggg gcgtgacagt
    20341 tattgcaggg gttcgtgaca gttagtacgg gagtgacggg cactggctgg caatgtctag
    20401 caacggcagg catttcggct gagggtaaaa gaactttccg ctaagcgata gactgtatgt
    20461 aaacacagta ttgcaaggac gcggaacatg cctcatgtgg cggccctcgc gagaattaat
    20521 tcagataaaa aaaatcctta gctttcgcta aggatgattt ctagcgatga ccctgctgat
    20581 tggttcgctg accgtagact ggttggcttg gtttcatcag ccatccgctt gccctcatct
    20641 gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
    20701 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
    20761 ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat
    20821 cctgtatatc gtgcgacatc gcgcgcgcgc gcgcggatgc tagcggcgcg ccaaaaaaaa
    20881 gcccgctcat ta
//
