LOCUS       sequence-486106-        9808 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Containing RP4 oriT and Cas9 module, for eliminating RP4 plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9808)
  AUTHORS   Tsoi R, Son HI, Hamrick GS, Tang K, Bethke JH, Lu J, Maddamsetti R, 
            You L
  TITLE     A predatory gene drive for targeted control of self-transmissible 
            plasmids.
  JOURNAL   Sci Adv. 2025 Apr 4;11(14):eads4735. doi: 10.1126/sciadv.ads4735. 
            Epub 2025 Apr 2.
  PUBMED    40173243
REFERENCE   2  (bases 1 to 9808)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2025-04-4"; volume: "11(14):eads4735. doi"; pages: " 
            10.1126/sciadv.ads4735. Epub 2025 Apr 2"
COMMENT     Sequence Label: DoS3 plasmid
FEATURES             Location/Qualifiers
     source          1..9808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(13..639)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     544..563
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        655..710
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    691..709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             717..728
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             735..4838
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             735..4838
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4839..4871
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag
                     /note="wild-type LAA variant"
                     /translation="AANDENYALAA"
     terminator      4898..4969
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(4975..4993,4994..4999,5000..5016,5017..5022,
                     5023..5048)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    4975..4993
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5000..5018
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        5069..5144
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             5165..5194
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5210..5227
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5283..5300)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5453..5499
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5524..6402)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        6429..6713
                     /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     6636..6655
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      7519..7605
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(7770..8358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7850..7869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(8446..8540)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(8561..9220)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     9135..9154
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(9221..9323)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            9377..9486
                     /note="incP origin of transfer"
     terminator      9752..9799
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ctcgaggcat gcttaagacc cactttcaca tttaagttgt ttttctaatc cgcatatgat
       61 caattcaagg ccgaataaga aggctggctc tgcaccttgg tgatcaaata attcgatagc
      121 ttgtcgtaat aatggcggca tactatcagt agtaggtgtt tccctttctt ctttagcgac
      181 ttgatgctct tgatcttcca atacgcaacc taaagtaaaa tgccccacag cgctgagtgc
      241 atataatgca ttctctagtg aaaaaccttg ttggcataaa aaggctaatt gattttcgag
      301 agtttcatac tgtttttctg taggccgtgt acctaaatgt acttttgctc catcgcgatg
      361 acttagtaaa gcacatctaa aacttttagc gttattacgt aaaaaatctt gccagctttc
      421 cccttctaaa gggcaaaagt gagtatggtg cctatctaac atctcaatgg ctaaggcgtc
      481 gagcaaagcc cgcttatttt ttacatgcca atacaatgta ggctgctcta cacctagctt
      541 ctgggcgagt ttacgggttg ttaaaccttc gattccgacc tcattaagca gctctaatgc
      601 gctgttaatc actttacttt tatctaatct agacatcatt aattcctaat ttttgttgac
      661 actctatcgt tgatagagtt attttaccac tccctatcag tgatagagaa agatctaaag
      721 aggagaaagg atctatggat aagaaatact caataggctt agatatcggc acaaatagcg
      781 tcggatgggc ggtgatcact gatgaatata aggttccgtc taaaaagttc aaggttctgg
      841 gaaatacaga ccgccacagt atcaaaaaaa atcttatagg ggctctttta tttgacagtg
      901 gagagacagc ggaagcgact cgtctcaaac ggacagctcg tagaaggtat acacgtcgga
      961 agaatcgtat ttgttatcta caggagattt tttcaaatga gatggcgaaa gtagatgata
     1021 gtttctttca tcgacttgaa gagtcttttt tggtggaaga agacaagaag catgaacgtc
     1081 atcctatttt tggaaatata gtagatgaag ttgcttatca tgagaaatat ccaactatct
     1141 atcatctgcg aaaaaaattg gtagattcta ctgataaagc ggatttgcgc ttaatctatt
     1201 tggccttagc gcatatgatt aagtttcgtg gtcatttttt gattgaggga gatttaaatc
     1261 ctgataatag tgatgtggac aaactattta tccagttggt acaaacctac aatcaattat
     1321 ttgaagaaaa ccctattaac gcaagtggag tagatgctaa agcgattctt tctgcacgat
     1381 tgagtaaatc aagacgatta gaaaatctca ttgctcagct ccccggtgag aagaaaaatg
     1441 gcttatttgg gaatctcatt gctttgtcat tgggtttgac ccctaatttt aaatcaaatt
     1501 ttgatttggc agaagatgct aaattacagc tttcaaaaga tacttacgat gatgatttag
     1561 ataatttatt ggcgcaaatt ggagatcaat atgctgattt gtttttggca gctaagaatt
     1621 tatcagatgc tattttactt tcagatatcc taagagtaaa tactgaaata actaaggctc
     1681 ccctatcagc ttcaatgatt aaacgctacg atgaacatca tcaagacttg actcttttaa
     1741 aagctttagt tcgacaacaa cttccagaaa agtataaaga aatctttttt gatcaatcaa
     1801 aaaacggata tgcaggttat attgatgggg gagctagcca agaagaattt tataaattta
     1861 tcaaaccaat tttagaaaaa atggatggta ctgaggaatt attggtgaaa ctaaatcgtg
     1921 aagatttgct gcgcaagcaa cggacctttg acaacggctc tattccccat caaattcact
     1981 tgggtgagct gcatgctatt ttgagaagac aagaagactt ttatccattt ttaaaagaca
     2041 atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc ttattatgtt ggtccattgg
     2101 cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc tgaagaaaca attaccccat
     2161 ggaattttga agaagttgtc gataaaggtg cttcagctca atcatttatt gaacgcatga
     2221 caaactttga taaaaatctt ccaaatgaaa aagtactacc aaaacatagt ttgctttatg
     2281 agtattttac ggtttataac gaattgacaa aggtcaaata tgttactgaa ggaatgcgaa
     2341 aaccagcatt tctttcaggt gaacagaaga aagccattgt tgatttactc ttcaaaacaa
     2401 atcgaaaagt aaccgttaag caattaaaag aagattattt caaaaaaata gaatgttttg
     2461 atagtgttga aatttcagga gttgaagata gatttaatgc ttcattaggt acctaccatg
     2521 atttgctaaa aattattaaa gataaagatt ttttggataa tgaagaaaat gaagatatct
     2581 tagaggatat tgttttaaca ttgaccttat ttgaagatag ggagatgatt gaggaaagac
     2641 ttaaaacata tgctcacctc tttgatgata aggtgatgaa acagcttaaa cgtcgccgtt
     2701 atactggttg gggacgtttg tctcgaaaat tgattaatgg tattagggat aagcaatctg
     2761 gcaaaacaat attagatttt ttgaaatcag atggttttgc caatcgcaat tttatgcagc
     2821 tgatccatga tgatagtttg acatttaaag aagacattca aaaagcacaa gtgtctggac
     2881 aaggcgatag tttacatgaa catattgcaa atttagctgg tagccctgct attaaaaaag
     2941 gtattttaca gactgtaaaa gttgttgatg aattggtcaa agtaatgggg cggcataagc
     3001 cagaaaatat cgttattgaa atggcacgtg aaaatcagac aactcaaaag ggccagaaaa
     3061 attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa agaattagga agtcagattc
     3121 ttaaagagca tcctgttgaa aatactcaat tgcaaaatga aaagctctat ctctattatc
     3181 tccaaaatgg aagagacatg tatgtggacc aagaattaga tattaatcgt ttaagtgatt
     3241 atgatgtcga tcacattgtt ccacaaagtt tccttaaaga cgattcaata gacaataagg
     3301 tcttaacgcg ttctgataaa aatcgtggta aatcggataa cgttccaagt gaagaagtag
     3361 tcaaaaagat gaaaaactat tggagacaac ttctaaacgc caagttaatc actcaacgta
     3421 agtttgataa tttaacgaaa gctgaacgtg gaggtttgag tgaacttgat aaagctggtt
     3481 ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa gcatgtggca caaattttgg
     3541 atagtcgcat gaatactaaa tacgatgaaa atgataaact tattcgagag gttaaagtga
     3601 ttaccttaaa atctaaatta gtttctgact tccgaaaaga tttccaattc tataaagtac
     3661 gtgagattaa caattaccat catgcccatg atgcgtatct aaatgccgtc gttggaactg
     3721 ctttgattaa gaaatatcca aaacttgaat cggagtttgt ctatggtgat tataaagttt
     3781 atgatgttcg taaaatgatt gctaagtctg agcaagaaat aggcaaagca accgcaaaat
     3841 atttctttta ctctaatatc atgaacttct tcaaaacaga aattacactt gcaaatggag
     3901 agattcgcaa acgccctcta atcgaaacta atggggaaac tggagaaatt gtctgggata
     3961 aagggcgaga ttttgccaca gtgcgcaaag tattgtccat gccccaagtc aatattgtca
     4021 agaaaacaga agtacagaca ggcggattct ccaaggagtc aattttacca aaaagaaatt
     4081 cggacaagct tattgctcgt aaaaaagact gggatccaaa aaaatatggt ggttttgata
     4141 gtccaacggt agcttattca gtcctagtgg ttgctaaggt ggaaaaaggg aaatcgaaga
     4201 agttaaaatc cgttaaagag ttactaggga tcacaattat ggaaagaagt tcctttgaaa
     4261 aaaatccgat tgacttttta gaagctaaag gatataagga agttaaaaaa gacttaatca
     4321 ttaaactacc taaatatagt ctttttgagt tagaaaacgg tcgtaaacgg atgctggcta
     4381 gtgccggaga attacaaaaa ggaaatgagc tggctctgcc aagcaaatat gtgaattttt
     4441 tatatttagc tagtcattat gaaaagttga agggtagtcc agaagataac gaacaaaaac
     4501 aattgtttgt ggagcagcat aagcattatt tagatgagat tattgagcaa atcagtgaat
     4561 tttctaagcg tgttatttta gcagatgcca atttagataa agttcttagt gcatataaca
     4621 aacatagaga caaaccaata cgtgaacaag cagaaaatat tattcattta tttacgttga
     4681 cgaatcttgg agctcccgct gcttttaaat attttgatac aacaattgat cgtaaacgat
     4741 atacgtctac aaaagaagtt ttagatgcca ctcttatcca tcaatccatc actggtcttt
     4801 atgaaacacg cattgatttg agtcagctag gaggtgacgc tgctaacgac gaaaactacg
     4861 ctctggctgc ttaactcgag taaggatctc caggcatcaa ataaaacgaa aggctcagtc
     4921 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcg atcttcccta
     4981 tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacat cagcaggacg
     5041 cactgaccca ttctgagaat agtgtatggt tttagagcta gaaatagcaa gttaaaataa
     5101 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgaagcttgg
     5161 gcccgaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc atcatcatca
     5221 tcatcattga gtttaaacgg tctccagctt ggctgttttg gcggatgaga gaagattttc
     5281 agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc
     5341 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     5401 gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa
     5461 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaactg
     5521 gatttatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg cacgaaactc
     5581 gctcgggctg gccccggtgc attttttaaa tactcgcgag aaatagagtt gatcgtcaaa
     5641 accaacattg cgaccgacgg tggcgatagg catccgggta gtgctcaaaa gcagcttcgc
     5701 ctgactaatg cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa
     5761 aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa
     5821 attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat
     5881 cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag
     5941 atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc
     6001 aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaaac ccgtattggc
     6061 aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca
     6121 ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg
     6181 gaacagcaaa atatcacccg gtcggcagac aaattctcgt ccctgatttt tcaccacccc
     6241 ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa
     6301 aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcgttaaa
     6361 cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccacc
     6421 attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta
     6481 ctggctcttc tcgctaaccc aaccggtaac cccgcttatt aaaagcattc tgtaacaaag
     6541 cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt
     6601 ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag
     6661 attagcggat cctacctgac gctttttatc gcaactctct actgtttctc cataaagatc
     6721 taaagaggag ggtacctata tgcatatgaa aaacatcaaa aaaaaccagg taatgaacct
     6781 gggtccgaac tctaaactgc tgaaagaata caaatcccag ctgatcgaac tgaacatcga
     6841 acagttcgaa gcaggtatcg gtctgatcct gggtgatgct tacatccgtt ctcgtgatga
     6901 aggtaaaacc tactgtatgc agttcgagtg gaaaaacaaa gcatacatgg accacgtatg
     6961 tctgctgtac gatcagtggg tactgtcccc gccgcacaaa aaagaacgtg ttaaccacct
     7021 gggtaacctg gtaatcacct ggggcgccca gactttcaaa caccaagctt tcaacaaact
     7081 ggctaacctg ttcatcgtta acaacaaaaa aaccatcccg aacaacctgg ttgaaaacta
     7141 cctgaccccg atgtctctgg catactggtt catggatgat ggtggtaaat gggattacaa
     7201 caaaaactct accaacaaat cgatcgtact gaacacccag tctttcactt tcgaagaagt
     7261 agaatacctg gttaagggtc tgcgtaacaa attccaactg aactgttacg taaaaatcaa
     7321 caaaaacaaa ccgatcatct acatcgattc tatgtcttac ctgatcttct acaacctgat
     7381 caaaccgtac ctgatcccgc agatgatgta caaactgccg aacactatct cctccgaaac
     7441 tttcctgaaa taatagggat aacagggtaa ttcagggccc gatcgatgcg gccgcttaat
     7501 taattaatct agaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     7561 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc
     7621 tagggcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7681 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7741 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7801 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7861 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7921 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7981 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     8041 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     8101 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     8161 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     8221 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     8281 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     8341 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     8401 aacgaaaact cacgttaagg gattttggtc atgactagtg cttggattct caccaataaa
     8461 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     8521 tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact
     8581 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     8641 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     8701 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     8761 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     8821 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     8881 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     8941 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     9001 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     9061 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     9121 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     9181 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     9241 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     9301 tggaacctct tacgtgccga tcaacgtctc aaccggtcag ccatccgctt gccctcatct
     9361 gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
     9421 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
     9481 ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat
     9541 cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag
     9601 cagggttatg cagcggaaaa gcgctgcttc cctgctgttt tgtggaatat ctaccgactg
     9661 gaaacaggca aatgcaggaa attactgaac tgaggggaca ggcgagagac gatcaacgtc
     9721 tcattttcgc cacgttaggc gtatcacgag gctagcataa ccccttgggg cctctaaacg
     9781 ggtcttgagg ggttttttgc gtcttcac
//
