LOCUS       sequence-489396-       11912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ISEre1 controled by pCas promoter, lambda-red 
            recombinase, SacB and ISEre1 guide RNA targeting pMB1 ori.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11912)
  AUTHORS   Zhou X, Yang S, Sun B, Dong F, Yin M, Jiang Y, Huang Z, Yang S
  TITLE     Implementation of RAGATH RNA-associated DNA Endonucleases as Genome 
            Editing Tool in Escherichia coli.
  JOURNAL   Biotechnol J. 2025 Mar;20(3):e70005. doi: 10.1002/biot.70005.
  PUBMED    40111000
REFERENCE   2  (bases 1 to 11912)
  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"; journalName: "Biotechnol 
            J."; date: "2025-03"; volume: "20(3):e70005. doi"; pages: " 
            10.1002/biot.70005"
COMMENT     Sequence Label: pISEre1
FEATURES             Location/Qualifiers
     source          1..11912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(107..1057)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1105..1327
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(1816..2631)
                     /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"
     CDS             complement(1816..2631)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2539..2558
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2623..2642
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(4269..5147)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5174..5458
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     5381..5400
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             5485..5493
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             5502..5918
                     /codon_start=1
                     /gene="gam"
                     /product="inhibitor of the host RecBCD nuclease in the 
                     lambda Red system"
                     /label=Gam
                     /note="protects the double-stranded DNA substrate for 
                     recombination"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             5924..6709
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             6706..7386
                     /codon_start=1
                     /gene="exo"
                     /product="5' to 3' double-stranded DNA exonuclease in the 
                     lambda Red system"
                     /label=Exo
                     /note="creates 3' single-stranded DNA tails"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFTKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      7387..7631
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             join(7968..8054,8055..9389)
                     /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      complement(9506..9553)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     9549..9567
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     ncRNA           complement(9606..9673)
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     promoter        complement(9878..9996)
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             10142..10978
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     CDS             11052..11900
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
ORIGIN
        1 catgggtatg gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt
       61 tgttagtctt gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt
      121 atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac
      181 cattgagatc atgcttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct
      241 gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat
      301 ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt
      361 cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc
      421 ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg
      481 gtttcaaaac ccattggtta agccttttaa actcatggta gttattttca agcattaaca
      541 tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta
      601 gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat
      661 gttccagatt atattttatg aattttttta actggaaaag ataaggcaat atctcttcac
      721 taaaaactaa ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct
      781 caaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg
      841 ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt
      901 tataagtgaa cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta
      961 gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg
     1021 ctagttcatt tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat
     1081 tttaatcact ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt
     1141 gagttgtggg tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta
     1201 gaccctctgt aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta
     1261 taatttatag aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt
     1321 ataactcact actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg
     1381 ctcctctaca aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg
     1441 gcaaatcgct gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt
     1501 cgtgacattc agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac
     1561 aggcgccttt tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg
     1621 ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt
     1681 cagcagttcc tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc
     1741 attcagactg gctaatgcac ccagtaaggc agcggtatca tcaacccact gcagcaacca
     1801 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     1861 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     1921 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     1981 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     2041 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     2101 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     2161 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     2221 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     2281 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     2341 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     2401 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattagcaa cgctaccttt
     2461 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     2521 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     2581 ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca taacacccct
     2641 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     2701 tgcaatgtaa catcagagat tttgagacac aacgtggctt tccctgcagc tgaacaatac
     2761 ttaatactat agaatgataa caaaataaac tactttttaa aagaattttg tgttataatc
     2821 tatttattat taagtattgg gtaatatttt ttgaagagat attttgaaaa agaaaaatta
     2881 aagcatatta aactaatttc ggaggtcatt aaaactatta ttgaaatcat caaactcatt
     2941 atggatttaa tttaaacttt ttattttagg aggcaaaaat gcgcaaactg ctgaaaagct
     3001 tcaaaaccga aatcaacccg accatcgaac agaaaatcaa aatcaacaaa accatcggca
     3061 cctgccgcta cgtgtacaac ttctacctgg gccacaacaa aaccctgtac gataacggcg
     3121 aaaaattcat gaccggcaaa agcttcagcg tgtggctgaa caacgaatac atcccgaaca
     3181 acccggataa aatctggatc aaagaagcct acagcaaggc cgtgaagaaa agcatcgaag
     3241 atggctgcac cgccttcacc cgcttcttca aacatcagag cgccttcccg aacttcaaaa
     3301 agaaaggcaa aagcgatgtg aaaatgtact tcgtgaaaaa caacaccaaa gattgccgct
     3361 gcgaacgcca ccgcctgaac atcccgaccc tgggctgggt gcgcatcaaa gaaaaaggct
     3421 acatcccgac caccaaagat ggctggaaaa tcaaaagcgg caccgtgagc gtgaaagccg
     3481 atcgctacta cgtgagcgtg ctggtggaaa tcccgaacgt gaaaatcgcc aacaacagca
     3541 acggcggcat cggcatcgat ctgggcctga aagatctggc catcgtgagc aacggcaaaa
     3601 cctacaaaaa catcaacaaa agcacccgca tcaaaaaact ggagaaaaaa ctgcgccgcg
     3661 aacagcgctg cctgagccgc aaatacgaga atctgaaaaa aggcgaaagc acacagaaaa
     3721 acattcagaa acagaaactg aaagtgcagc gcctgcacca caaaatcgat aacatccgca
     3781 ccgattacat caataaaagc atcgccgaaa tcgtgaaaac caaaccgagc tacatcacca
     3841 tcgaaaacct gaacgtgagc ggcatgatga aaaaccgcca cctgagcaaa gccgtggcga
     3901 gccaaaaatt ctacgaattc cgcaccaaac tgaaagccaa atgcgatgaa aacggcatcg
     3961 aactgcgcgt ggtggatcgc tggtacccga gcagcaaaat ctgccactgc tgcggcgcca
     4021 tcaaaaaaga tctgaaactg agcgatcgca tctaccgctg cgattgcggc tacgtggaag
     4081 atcgcgattt caacgccgcc ctgaacctgc gcgatgccct gacctacgaa gtggcctgaa
     4141 gtatatttta gatgaagatt atttcttaat ctagacatga gcggatacat atttgaatgt
     4201 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgca
     4261 tcgatttatt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     4321 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaagta gagttgatcg
     4381 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     4441 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     4501 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     4561 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     4621 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     4681 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     4741 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     4801 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     4861 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     4921 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     4981 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     5041 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     5101 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     5161 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
     5221 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
     5281 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
     5341 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
     5401 ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac
     5461 ccgttttttt gggaattcga gctctaagga ggttataaaa aatggatatt aatactgaaa
     5521 ctgagatcaa gcaaaagcat tcactaaccc cctttcctgt tttcctaatc agcccggcat
     5581 ttcgcgggcg atattttcac agctatttca ggagttcagc catgaacgct tattacattc
     5641 aggatcgtct tgaggctcag agctgggcgc gtcactacca gcagctcgcc cgtgaagaga
     5701 aagaggcaga actggcagac gacatggaaa aaggcctgcc ccagcacctg tttgaatcgc
     5761 tatgcatcga tcatttgcaa cgccacgggg ccagcaaaaa atccattacc cgtgcgtttg
     5821 atgacgatgt tgagtttcag gagcgcatgg cagaacacat ccggtacatg gttgaaacca
     5881 ttgctcacca ccaggttgat attgattcag aggtataaaa cgaatgagta ctgcactcgc
     5941 aacgctggct gggaagctgg ctgaacgtgt cggcatggat tctgtcgacc cacaggaact
     6001 gatcaccact cttcgccaga cggcatttaa aggtgatgcc agcgatgcgc agttcatcgc
     6061 attactgatc gttgccaacc agtacggcct taatccgtgg acgaaagaaa tttacgcctt
     6121 tcctgataag cagaatggca tcgttccggt ggtgggcgtt gatggctggt cccgcatcat
     6181 caatgaaaac cagcagtttg atggcatgga ctttgagcag gacaatgaat cctgtacatg
     6241 ccggatttac cgcaaggacc gtaatcatcc gatctgcgtt accgaatgga tggatgaatg
     6301 ccgccgcgaa ccattcaaaa ctcgcgaagg cagagaaatc acggggccgt ggcagtcgca
     6361 tcccaaacgg atgttacgtc ataaagccat gattcagtgt gcccgtctgg ccttcggatt
     6421 tgctggtatc tatgacaagg atgaagccga gcgcattgtc gaaaatactg catacactgc
     6481 agaacgtcag ccggaacgcg acatcactcc ggttaacgat gaaaccatgc aggagattaa
     6541 cactctgctg atcgccctgg ataaaacatg ggatgacgac ttattgccgc tctgttccca
     6601 gatatttcgc cgcgacattc gtgcatcgtc agaactgaca caggccgaag cagtaaaagc
     6661 tcttggattc ctgaaacaga aagccgcaga gcagaaggtg gcagcatgac accggacatt
     6721 atcctgcagc gtaccgggat cgatgtgaga gctgtcgaac agggggatga tgcgtggcac
     6781 aaattacggc tcggcgtcat caccgcttca gaagttcaca acgtgatagc aaaaccccgc
     6841 tccggaaaga agtggcctga catgaaaatg tcctacttcc acaccctgct tgctgaggtt
     6901 tgcaccggtg tggctccgga agttaacgct aaagcactgg cctggggaaa acagtacgag
     6961 aacgacgcca gaaccctgtt tgaattcact tccggcgtga atgttactga atccccgatc
     7021 atctatcgcg acgaaagtat gcgtaccgcc tgctctcccg atggtttatg cagtgacggc
     7081 aacggccttg aactgaaatg cccgtttacc tcccgggatt tcacgaagtt ccggctcggt
     7141 ggtttcgagg ccataaagtc agcttacatg gcccaggtgc agtacagcat gtgggtgacg
     7201 cgaaaaaatg cctggtactt tgccaactat gacccgcgta tgaagcgtga aggcctgcat
     7261 tatgtcgtga ttgagcggga tgaaaagtac atggcgagtt ttgacgagat cgtgccggag
     7321 ttcatcgaaa aaatggacga ggcactggct gaaattggtt ttgtatttgg ggagcaatgg
     7381 cgatgacgca tcctcacgat aatatccggg taggcgcaat cactttcgtc tactccgtta
     7441 caaagcgagg ctgggtattt cccggccttt ctgttatccg aaatccactg aaagcacagc
     7501 ggctggctga ggagataaat aataaacgag gggctgtatg cacaaagcat cttctgttga
     7561 gttaagaacg agtatcgaga tggcacatag ccttgctcaa attggaatca ggtttgtgcc
     7621 aataccagta gaaacagacg aagaatccat gggtatggac agatcctctt taggcccgta
     7681 gtctgcaaat ccttttatga ttttctatca aacaaaagag gaaaatagac cagttgcaat
     7741 ccaaacgaga gtctaataga atgaggtcga aaagtaaatc gcgcgggttt gttactgata
     7801 aagcaggcaa gacctaaaat gtgtaaaggg caaagtgtat actttggcgt caccccttac
     7861 atattttagg tcttttttta ttgtgcgtaa ctaacttgcc atcttcaaac aggagggctg
     7921 gaagaagcag accgctaaca cagtacataa aaaaggagac atgaacgatg aacatcaaaa
     7981 agtttgcaaa acaagcaaca gtattaacct ttactaccgc actgctggca ggaggcgcaa
     8041 ctcaagcgtt tgcgaaagaa acgaaccaaa agccatataa ggaaacatac ggcatttccc
     8101 atattacacg ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa aaatatcaag
     8161 ttcctgaatt cgattcgtcc acaattaaaa atatctcttc tgcaaaaggc ctggacgttt
     8221 gggacagctg gccattacaa aacgctgacg gcactgtcgc aaactatcac ggctaccaca
     8281 tcgtctttgc attagccgga gatcctaaaa atgcggatga cacatcgatt tacatgttct
     8341 atcaaaaagt cggcgaaact tctattgaca gctggaaaaa cgctggccgc gtctttaaag
     8401 acagcgacaa attcgatgca aatgattcta tcctaaaaga ccaaacacaa gaatggtcag
     8461 gttcagccac atttacatct gacggaaaaa tccgtttatt ctacactgat ttctccggta
     8521 aacattacgg caaacaaaca ctgacaactg cacaagttaa cgtatcagca tcagacagct
     8581 ctttgaacat caacggtgta gaggattata aatcaatctt tgacggtgac ggaaaaacgt
     8641 atcaaaatgt acagcagttc atcgatgaag gcaactacag ctcaggcgac aaccatacgc
     8701 tgagagatcc tcactacgta gaagataaag gccacaaata cttagtattt gaagcaaaca
     8761 ctggaactga agatggctac caaggcgaag aatctttatt taacaaagca tactatggca
     8821 aaagcacatc attcttccgt caagaaagtc aaaaacttct gcaaagcgat aaaaaacgca
     8881 cggctgagtt agcaaacggc gctctcggta tgattgagct aaacgatgat tacacactga
     8941 aaaaagtgat gaaaccgctg attgcatcta acacagtaac agatgaaatt gaacgcgcga
     9001 acgtctttaa aatgaacggc aaatggtacc tgttcactga ctcccgcgga tcaaaaatga
     9061 cgattgacgg cattacgtct aacgatattt acatgcttgg ttatgtttct aattctttaa
     9121 ctggcccata caagccgctg aacaaaactg gccttgtgtt aaaaatggat cttgatccta
     9181 acgatgtaac ctttacttac tcacacttcg ctgtacctca agcgaaagga aacaatgtcg
     9241 tgattacaag ctatatgaca aacagaggat tctacgcaga caaacaatca acgtttgcgc
     9301 caagcttcct gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac agcatccttg
     9361 aacaaggaca attaacagtt aacaaataaa aacgcaaaag aaaatgccga tatcctattg
     9421 gcattttctt ttatttctta tcaacataaa ggtgaatccc atatgaacta tataaaagca
     9481 ggcaaatggc taaccgtatt cctcacaaaa aacccctcaa gacccgttta gaggccccaa
     9541 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
     9601 tgttagtccc attcgccatg ccgaagcatg ttgcccagcc ggcgccagcg aggaggctgg
     9661 gaccatgccg gcctggaagc tccctcgtgc gctctgccac tcaaaatgtg ttcaaatata
     9721 ctcaaacaca tcgagaaatt aaagtccttg cggatacttc ttactgcttc gatgtgtatg
     9781 ctttggcgat tgtaaacaat acgctactca caagttcgtg tacactccac agtcgtaaat
     9841 tcctgcgata gcccgcagta catacttacg tttgcgttac gaccagtcta aaaagcgcct
     9901 gaattcgcga ccttctcgtt actgacagga aaatgggcca ttggcaacca gggaaagatg
     9961 aacgtgatga tgttcacaat ttgctgaatt gtggtgatgt gatgctcacc gcatttcctg
    10021 aaaattcacg ctgtatcttg aaaaatcgac gttttttacg tggttttccg tcgaaaattt
    10081 aaggtaagaa cctgacctcg tgattactat ttcgccgtgt tgacgacatc aggaggccag
    10141 tatgaccgta ttacatagtg tggatttttt tccgtctggt aacgcgtccg tggcgataga
    10201 accccggctc ccgcaggcgg attttcctga acatcatcat gattttcatg aaattgtgat
    10261 tgtcgaacat ggcacgggta ttcatgtgtt taatgggcag ccctatacca tcaccggtgg
    10321 cacggtctgt ttcgtacgcg atcatgatcg gcatctgtat gaacataccg ataatctgtg
    10381 tctgaccaat gtgctgtatc gctcgccgga tcgatttcag tttctcgccg ggctgaatca
    10441 gttgctgcca caagagctgg atgggcagta tccgtctcac tggcgcgtta accacagcgt
    10501 attgcagcag gtgcgacagc tggttgcaca gatggaacag caggaagggg aaaatgattt
    10561 accctcgacc gccagtcgcg agatcttgtt tatgcaatta ctgctcttgc tgcgtaaaag
    10621 cagtttgcag gagaacctgg aaaacagcgc atcacgtctc aacttgcttc tggcctggct
    10681 ggaggaccat tttgccgatg aggtgaattg ggatgccgtg gcggatcaat tttctctttc
    10741 actgcgtacg ctacatcggc agcttaagca gcaaacggga ctgacgcctc agcgatacct
    10801 gaaccgcctg cgactgatga aagcccgaca tctgctacgc cacagcgagg ccagcgttac
    10861 tgacatcgcc tatcgctgtg gattcagcga cagtaaccac ttttcgacgc tttttcgccg
    10921 agagtttaac tggtcaccgc gtgatattcg ccagggacgg gatggctttc tgcaataacg
    10981 cgaatcttct caacgtattt gtacgccata ttgcgaataa tcaacttcgt tctctggccg
    11041 aggtagccac ggtggcgcat cagttaaaac ttctcaaaga tgattttttt gccagcgacc
    11101 agcaggcagt cgctgtggct gaccgttatc cgcaagatgt ctttgctgaa catacacatg
    11161 atttttgtga gctggtgatt gtctggcgcg gtaatggcct gcatgtactc aacgatcgcc
    11221 cttatcgcat tacccgtggc gatctctttt acattcatgc tgacgataaa cactcctacg
    11281 cttccgttaa cgatctggtt ttgcagaata ttatttattg cccggagcgt ctgaagctga
    11341 atcttgactg gcagggggcg attccgggat ttaacgccag cgcagggcaa ccacactggc
    11401 gcttaggtag catggggatg gcgcaggcgc ggcaggttat cggtcagctt gagcatgaaa
    11461 gtagtcagca tgtgccgttt gctaacgaaa tggctgagtt gctgttcggg cagttggtga
    11521 tgttgctgaa tcgccatcgt tacaccagtg attcgttgcc gccaacatcc agcgaaacgt
    11581 tgctggataa gctgattacc cggctggcgg ctagcctgaa aagtcccttt gcgctggata
    11641 aattttgtga tgaggcatcg tgcagtgagc gcgttttgcg tcagcaattt cgccagcaga
    11701 ctggaatgac catcaatcaa tatctgcgac aggtcagagt gtgtcatgcg caatatcttc
    11761 tccagcatag ccgcctgtta atcagtgata tttcgaccga atgtggcttt gaagatagta
    11821 actatttttc ggtggtgttt acccgggaaa ccgggatgac gcccagccag tggcgtcatc
    11881 tcaattcgca gaaagattaa ttcgccatgc cc
//
