LOCUS       40924_16710       14605 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Constitutive expression of cas9 and inducible expression of lambda 
            RED and sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14605)
  AUTHORS   Li , Q i, Sun, Bingbing, Chen, Jun, Zhang, Yiwen, Jiang, Y u Jiang, 
            Yang, Sheng
  TITLE     A modified pCas/pTargetF system for CRISPR-Cas9-assisted genome 
            editing in Escherichia coli
  JOURNAL   Acta Biochimica et Biophysica Sinica, 2021
REFERENCE   2  (bases 1 to 14605)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 14605)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Acta
            Biochimica et Biophysica Sinica, 2021"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..14605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(110..1057)
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(1105..1327)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(1835..2647)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2995..7098
                     /codon_start=1
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7234..8109)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        8136..8420
                     /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)"
     RBS             8447..8455
                     /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             8464..8877
                     /codon_start=1
                     /label=Gam
                     /note="inhibitor of the host RecBCD nuclease in the lambda 
                     Red system"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             8886..9668
                     /codon_start=1
                     /label=Beta
                     /note="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             9668..10345
                     /codon_start=1
                     /label=Exo
                     /note="5' to 3' double-stranded DNA exonuclease in the 
                     lambda
                     Red system"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      10349..10593
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             10930..12348
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     misc_RNA        complement(12474..12549)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(12571..12689)
                     /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             12835..13668
                     /codon_start=1
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     CDS             13745..14590
                     /codon_start=1
                     /label=rhaR
                     /note="transcriptional activator of rhaR and rhaS"
                     /translation="VAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     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 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct
      241 gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttacgt 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 ttgcggccgc aatcgggcaa atcgctgaat attccttttg tctccgacca tcaggcacct
     1501 gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg
     1561 gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac ccataataca
     1621 agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat
     1681 ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat
     1741 tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa
     1801 cccactgcag caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
     1861 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     1921 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     1981 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     2041 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
     2101 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     2161 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     2221 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     2281 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     2341 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     2401 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     2461 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     2521 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     2581 aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat
     2641 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     2701 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttccc
     2761 tgcagctgaa caatacttaa tactatagaa tgataacaaa ataaactact ttttaaaaga
     2821 attttgtgtt ataatctatt tattattaag tattgggtaa tattttttga agagatattt
     2881 tgaaaaagaa aaattaaagc atattaaact aatttcggag gtcattaaaa ctattattga
     2941 aatcatcaaa ctcattatgg atttaattta aactttttat tttaggaggc aaaaatggat
     3001 aagaaatact caataggctt agatatcggc acaaatagcg tcggatgggc ggtgatcact
     3061 gatgattata aggttccgtc taaaaagttc aaggttctgg gaaatacaga ccgccacagt
     3121 atcaaaaaaa atcttatagg ggctctttta tttgacagtg gagagacagc ggaagcgact
     3181 cgtctcaaac ggacagctcg tagaaggtat acacgtcgga agaatcgtat ttgttatcta
     3241 caggagattt tttcaaatga gatggcgaaa gtagatgata gtttctttca tcgacttgaa
     3301 gagtcttttt tggtggaaga agacaagaag catgaacgtc atcctatttt tggaaatata
     3361 gtagatgaag ttgcttatca tgagaaatat ccaactatct atcatctgcg aaaaaaattg
     3421 gtagattcta ctgataaagc ggatttgcgc ttaatctatt tggccttagc gcatatgatt
     3481 aagtttcgtg gtcatttttt gattgaggga gatttaaatc ctgataatag tgatgtggac
     3541 aaactattta tccagttggt acaaacctac aatcaattat ttgaagaaaa ccctattaac
     3601 gcaagtggag tagatgctaa agcgattctt tctgcacgat tgagtaaatc aagacgatta
     3661 gaaaatctca ttgctcagct ccccggtgag aagaaaaatg gcttatttgg gaatctcatt
     3721 gctttgtcat tgggtttgac ccctaatttt aaatcaaatt ttgatttggc agaagatgct
     3781 aaattacagc tttcaaaaga tacttacgat gatgatttag ataatttatt ggcgcaaatt
     3841 ggagatcaat atgctgattt gtttttggca gctaagaatt tatcagatgc tattttactt
     3901 tcagatatcc taagagtaaa tactgaaata actaaggctc ccctatcagc ttcaatgatt
     3961 aaacgctacg atgaacatca tcaagacttg actcttttaa aagctttagt tcgacaacaa
     4021 cttccagaaa agtataaaga aatctttttt gatcaatcaa aaaacggata tgcaggttat
     4081 attgatgggg gagctagcca agaagaattt tataaattta tcaaaccaat tttagaaaaa
     4141 atggatggta ctgaggaatt attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa
     4201 cggacctttg acaacggctc tattccccat caaattcact tgggtgagct gcatgctatt
     4261 ttgagaagac aagaagactt ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa
     4321 atcttgactt ttcgaattcc ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt
     4381 gcatggatga ctcggaagtc tgaagaaaca attaccccat ggaattttga agaagttgtc
     4441 gataaaggtg cttcagctca atcatttatt gaacgcatga caaactttga taaaaatctt
     4501 ccaaatgaaa aagtactacc aaaacatagt ttgctttatg agtattttac ggtttataac
     4561 gaattgacaa aggtcaaata tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt
     4621 gaacagaaga aagccattgt tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag
     4681 caattaaaag aagattattt caaaaaaata gaatgttttg atagtgttga aatttcagga
     4741 gttgaagata gatttaatgc ttcattaggt acctaccatg atttgctaaa aattattaaa
     4801 gataaagatt ttttggataa tgaagaaaat gaagatatct tagaggatat tgttttaaca
     4861 ttgaccttat ttgaagatag ggagatgatt gaggaaagac ttaaaacata tgctcacctc
     4921 tttgatgata aggtgatgaa acagcttaaa cgtcgccgtt atactggttg gggacgtttg
     4981 tctcgaaaat tgattaatgg tattagggat aagcaatctg gcaaaacaat attagatttt
     5041 ttgaaatcag atggttttgc caatcgcaat tttatgcagc tgatccatga tgatagtttg
     5101 acatttaaag aagacattca aaaagcacaa gtgtctggac aaggcgatag tttacatgaa
     5161 catattgcaa atttagctgg tagccctgct attaaaaaag gtattttaca gactgtaaaa
     5221 gttgttgatg aattggtcaa agtaatgggg cggcataagc cagaaaatat cgttattgaa
     5281 atggcacgtg aaaatcagac aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa
     5341 cgaatcgaag aaggtatcaa agaattagga agtcagattc ttaaagagca tcctgttgaa
     5401 aatactcaat tgcaaaatga aaagctctat ctctattatc tccaaaatgg aagagacatg
     5461 tatgtggacc aagaattaga tattaatcgt ttaagtgatt atgatgtcga tcacattgtt
     5521 ccacaaagtt tccttaaaga cgattcaata gacaataagg tcttaacgcg ttctgataaa
     5581 aatcgtggta aatcggataa cgttccaagt gaagaagtag tcaaaaagat gaaaaactat
     5641 tggagacaac ttctaaacgc caagttaatc actcaacgta agtttgataa tttaacgaaa
     5701 gctgaacgtg gaggtttgag tgaacttgat aaagctggtt ttatcaaacg ccaattggtt
     5761 gaaactcgcc aaatcactaa gcatgtggca caaattttgg atagtcgcat gaatactaaa
     5821 tacgatgaaa atgataaact tattcgagag gttaaagtga ttaccttaaa atctaaatta
     5881 gtttctgact tccgaaaaga tttccaattc tataaagtac gtgagattaa caattaccat
     5941 catgcccatg atgcgtatct aaatgccgtc gttggaactg ctttgattaa gaaatatcca
     6001 aaacttgaat cggagtttgt ctatggtgat tataaagttt atgatgttcg taaaatgatt
     6061 gctaagtctg agcaagaaat aggcaaagca accgcaaaat atttctttta ctctaatatc
     6121 atgaacttct tcaaaacaga aattacactt gcaaatggag agattcgcaa acgccctcta
     6181 atcgaaacta atggggaaac tggagaaatt gtctgggata aagggcgaga ttttgccaca
     6241 gtgcgcaaag tattgtccat gccccaagtc aatattgtca agaaaacaga agtacagaca
     6301 ggcggattct ccaaggagtc aattttacca aaaagaaatt cggacaagct tattgctcgt
     6361 aaaaaagact gggatccaaa aaaatatggt ggttttgata gtccaacggt agcttattca
     6421 gtcctagtgg ttgctaaggt ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag
     6481 ttactaggga tcacaattat ggaaagaagt tcctttgaaa aaaatccgat tgacttttta
     6541 gaagctaaag gatataagga agttaaaaaa gacttaatca ttaaactacc taaatatagt
     6601 ctttttgagt tagaaaacgg tcgtaaacgg atgctggcta gtgccggaga attacaaaaa
     6661 ggaaatgagc tggctctgcc aagcaaatat gtgaattttt tatatttagc tagtcattat
     6721 gaaaagttga agggtagtcc agaagataac gaacaaaaac aattgtttgt ggagcagcat
     6781 aagcattatt tagatgagat tattgagcaa atcagtgaat tttctaagcg tgttatttta
     6841 gcagatgcca atttagataa agttcttagt gcatataaca aacatagaga caaaccaata
     6901 cgtgaacaag cagaaaatat tattcattta tttacgttga cgaatcttgg agctcccgct
     6961 gcttttaaat attttgatac aacaattgat cgtaaacgat atacgtctac aaaagaagtt
     7021 ttagatgcca ctcttatcca tcaatccatc actggtcttt atgaaacacg cattgatttg
     7081 agtcagctag gaggtgactg aagtatattt tagatgaaga ttatttctta atctagacat
     7141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7201 tccccgaaaa gtgccacctg catcgattta ttatgacaac ttgacggcta catcattcac
     7261 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac
     7321 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat
     7381 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa
     7441 gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac
     7501 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg
     7561 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat
     7621 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc
     7681 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc
     7741 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca
     7801 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg
     7861 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa
     7921 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc
     7981 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt
     8041 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc
     8101 ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc catattgcat
     8161 cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac cggtaacccc
     8221 gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa
     8281 aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg tcacactttg
     8341 ctatgccata gcatttttat ccataagatt agcggatcct acctgacgct ttttatcgca
     8401 actctctact gtttctccat acccgttttt ttgggaattc gagctctaag gaggttataa
     8461 aaaatggata ttaatactga aactgagatc aagcaaaagc attcactaac cccctttcct
     8521 gttttcctaa tcagcccggc atttcgcggg cgatattttc acagctattt caggagttca
     8581 gccatgaacg cttattacat tcaggatcgt cttgaggctc agagctgggc gcgtcactac
     8641 cagcagctcg cccgtgaaga gaaagaggca gaactggcag acgacatgga aaaaggcctg
     8701 ccccagcacc tgtttgaatc gctatgcatc gatcatttgc aacgccacgg ggccagcaaa
     8761 aaatccatta cccgtgcgtt tgatgacgat gttgagtttc aggagcgcat ggcagaacac
     8821 atccggtaca tggttgaaac cattgctcac caccaggttg atattgattc agaggtataa
     8881 aacgaatgag tactgcactc gcaacgctgg ctgggaagct ggctgaacgt gtcggcatgg
     8941 attctgtcga cccacaggaa ctgatcacca ctcttcgcca gacggcattt aaaggtgatg
     9001 ccagcgatgc gcagttcatc gcattactga tcgttgccaa ccagtacggc cttaatccgt
     9061 ggacgaaaga aatttacgcc tttcctgata agcagaatgg catcgttccg gtggtgggcg
     9121 ttgatggctg gtcccgcatc atcaatgaaa accagcagtt tgatggcatg gactttgagc
     9181 aggacaatga atcctgtaca tgccggattt accgcaagga ccgtaatcat ccgatctgcg
     9241 ttaccgaatg gatggatgaa tgccgccgcg aaccattcaa aactcgcgaa ggcagagaaa
     9301 tcacggggcc gtggcagtcg catcccaaac ggatgttacg tcataaagcc atgattcagt
     9361 gtgcccgtct ggccttcgga tttgctggta tctatgacaa ggatgaagcc gagcgcattg
     9421 tcgaaaatac tgcatacact gcagaacgtc agccggaacg cgacatcact ccggttaacg
     9481 atgaaaccat gcaggagatt aacactctgc tgatcgccct ggataaaaca tgggatgacg
     9541 acttattgcc gctctgttcc cagatatttc gccgcgacat tcgtgcatcg tcagaactga
     9601 cacaggccga agcagtaaaa gctcttggat tcctgaaaca gaaagccgca gagcagaagg
     9661 tggcagcatg acaccggaca ttatcctgca gcgtaccggg atcgatgtga gagctgtcga
     9721 acagggggat gatgcgtggc acaaattacg gctcggcgtc atcaccgctt cagaagttca
     9781 caacgtgata gcaaaacccc gctccggaaa gaagtggcct gacatgaaaa tgtcctactt
     9841 ccacaccctg cttgctgagg tttgcaccgg tgtggctccg gaagttaacg ctaaagcact
     9901 ggcctgggga aaacagtacg agaacgacgc cagaaccctg tttgaattca cttccggcgt
     9961 gaatgttact gaatccccga tcatctatcg cgacgaaagt atgcgtaccg cctgctctcc
    10021 cgatggttta tgcagtgacg gcaacggcct tgaactgaaa tgcccgttta cctcccggga
    10081 tttcatgaag ttccggctcg gtggtttcga ggccataaag tcagcttaca tggcccaggt
    10141 gcagtacagc atgtgggtga cgcgaaaaaa tgcctggtac tttgccaact atgacccgcg
    10201 tatgaagcgt gaaggcctgc attatgtcgt gattgagcgg gatgaaaagt acatggcgag
    10261 ttttgacgag atcgtgccgg agttcatcga aaaaatggac gaggcactgg ctgaaattgg
    10321 ttttgtattt ggggagcaat ggcgatgacg catcctcacg ataatatccg ggtaggcgca
    10381 atcactttcg tctactccgt tacaaagcga ggctgggtat ttcccggcct ttctgttatc
    10441 cgaaatccac tgaaagcaca gcggctggct gaggagataa ataataaacg aggggctgta
    10501 tgcacaaagc atcttctgtt gagttaagaa cgagtatcga gatggcacat agccttgctc
    10561 aaattggaat caggtttgtg ccaataccag tagaaacaga cgaagaatcc atgggtatgg
    10621 acagatcctc tttaggcccg tagtctgcaa atccttttat gattttctat caaacaaaag
    10681 aggaaaatag accagttgca atccaaacga gagtctaata gaatgaggtc gaaaagtaaa
    10741 tcgcgcgggt ttgttactga taaagcaggc aagacctaaa atgtgtaaag ggcaaagtgt
    10801 atactttggc gtcacccctt acatatttta ggtctttttt tattgtgcgt aactaacttg
    10861 ccatcttcaa acaggagggc tggaagaagc agaccgctaa cacagtacat aaaaaaggag
    10921 acatgaacga tgaacatcaa aaagtttgca aaacaagcaa cagtattaac ctttactacc
    10981 gcactgctgg caggaggcgc aactcaagcg tttgcgaaag aaacgaacca aaagccatat
    11041 aaggaaacat acggcatttc ccatattaca cgccatgata tgctgcaaat ccctgaacag
    11101 caaaaaaatg aaaaatatca agttcctgaa ttcgattcgt ccacaattaa aaatatctct
    11161 tctgcaaaag gcctggacgt ttgggacagc tggccattac aaaacgctga cggcactgtc
    11221 gcaaactatc acggctacca catcgtcttt gcattagccg gagatcctaa aaatgcggat
    11281 gacacatcga tttacatgtt ctatcaaaaa gtcggcgaaa cttctattga cagctggaaa
    11341 aacgctggcc gcgtctttaa agacagcgac aaattcgatg caaatgattc tatcctaaaa
    11401 gaccaaacac aagaatggtc aggttcagcc acatttacat ctgacggaaa aatccgttta
    11461 ttctacactg atttctccgg taaacattac ggcaaacaaa cactgacaac tgcacaagtt
    11521 aacgtatcag catcagacag ctctttgaac atcaacggtg tagaggatta taaatcaatc
    11581 tttgacggtg acggaaaaac gtatcaaaat gtacagcagt tcatcgatga aggcaactac
    11641 agctcaggcg acaaccatac gctgagagat cctcactacg tagaagataa aggccacaaa
    11701 tacttagtat ttgaagcaaa cactggaact gaagatggct accaaggcga agaatcttta
    11761 tttaacaaag catactatgg caaaagcaca tcattcttcc gtcaagaaag tcaaaaactt
    11821 ctgcaaagcg ataaaaaacg cacggctgag ttagcaaacg gcgctctcgg tatgattgag
    11881 ctaaacgatg attacacact gaaaaaagtg atgaaaccgc tgattgcatc taacacagta
    11941 acagatgaaa ttgaacgcgc gaacgtcttt aaaatgaacg gcaaatggta cctgttcact
    12001 gactcccgcg gatcaaaaat gacgattgac ggcattacgt ctaacgatat ttacatgctt
    12061 ggttatgttt ctaattcttt aactggccca tacaagccgc tgaacaaaac tggccttgtg
    12121 ttaaaaatgg atcttgatcc taacgatgta acctttactt actcacactt cgctgtacct
    12181 caagcgaaag gaaacaatgt cgtgattaca agctatatga caaacagagg attctacgca
    12241 gacaaacaat caacgtttgc gccaagcttc ctgctgaaca tcaaaggcaa gaaaacatct
    12301 gttgtcaaag acagcatcct tgaacaagga caattaacag ttaacaaata aaaacgcaaa
    12361 agaaaatgcc gatatcctat tggcattttc ttttatttct tatcaacata aaggtgaatc
    12421 ccatatgaac tatataaaag caggcaaatg gctaaccgta ttcctcaaaa aaagcaccga
    12481 ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag
    12541 ctctaaaacg gttggactca agacgatagt tacgaccagt ctaaaaagcg cctgaattcg
    12601 cgaccttctc gttactgaca ggaaaatggg ccattggcaa ccagggaaag atgaacgtga
    12661 tgatgttcac aatttgctga attgtggtga tgtgatgctc accgcatttc ctgaaaattc
    12721 acgctgtatc ttgaaaaatc gacgtttttt acgtggtttt ccgtcgaaaa tttaaggtaa
    12781 gaacctgacc tcgtgattac tatttcgccg tgttgacgac atcaggaggc cagtatgacc
    12841 gtattacata gtgtggattt ttttccgtct ggtaacgcgt ccgtggcgat agaaccccgg
    12901 ctcccgcagg cggattttcc tgaacatcat catgattttc atgaaattgt gattgtcgaa
    12961 catggcacgg gtattcatgt gtttaatggg cagccctata ccatcaccgg tggcacggtc
    13021 tgtttcgtac gcgatcatga tcggcatctg tatgaacata ccgataatct gtgtctgacc
    13081 aatgtgctgt atcgctcgcc ggatcgattt cagtttctcg ccgggctgaa tcagttgctg
    13141 ccacaagagc tggatgggca gtatccgtct cactggcgcg ttaaccacag cgtattgcag
    13201 caggtgcgac agctggttgc acagatggaa cagcaggaag gggaaaatga tttaccctcg
    13261 accgccagtc gcgagatctt gtttatgcaa ttactgctct tgctgcgtaa aagcagtttg
    13321 caggagaacc tggaaaacag cgcatcacgt ctcaacttgc ttctggcctg gctggaggac
    13381 cattttgccg atgaggtgaa ttgggatgcc gtggcggatc aattttctct ttcactgcgt
    13441 acgctacatc ggcagcttaa gcagcaaacg ggactgacgc ctcagcgata cctgaaccgc
    13501 ctgcgactga tgaaagcccg acatctgcta cgccacagcg aggccagcgt tactgacatc
    13561 gcctatcgct gtggattcag cgacagtaac cacttttcga cgctttttcg ccgagagttt
    13621 aactggtcac cgcgtgatat tcgccaggga cgggatggct ttctgcaata acgcgaatct
    13681 tctcaacgta tttgtacgcc atattgcgaa taatcaactt cgttctctgg ccgaggtagc
    13741 cacggtggcg catcagttaa aacttctcaa agatgatttt tttgccagcg accagcaggc
    13801 agtcgctgtg gctgaccgtt atccgcaaga tgtctttgct gaacatacac atgatttttg
    13861 tgagctggtg attgtctggc gcggtaatgg cctgcatgta ctcaacgatc gcccttatcg
    13921 cattacccgt ggcgatctct tttacattca tgctgacgat aaacactcct acgcttccgt
    13981 taacgatctg gttttgcaga atattattta ttgcccggag cgtctgaagc tgaatcttga
    14041 ctggcagggg gcgattccgg gatttaacgc cagcgcaggg caaccacact ggcgcttagg
    14101 tagcatgggg atggcgcagg cgcggcaggt tatcggtcag cttgagcatg aaagtagtca
    14161 gcatgtgccg tttgctaacg aaatggctga gttgctgttc gggcagttgg tgatgttgct
    14221 gaatcgccat cgttacacca gtgattcgtt gccgccaaca tccagcgaaa cgttgctgga
    14281 taagctgatt acccggctgg cggctagcct gaaaagtccc tttgcgctgg ataaattttg
    14341 tgatgaggca tcgtgcagtg agcgcgtttt gcgtcagcaa tttcgccagc agactggaat
    14401 gaccatcaat caatatctgc gacaggtcag agtgtgtcat gcgcaatatc ttctccagca
    14461 tagccgcctg ttaatcagtg atatttcgac cgaatgtggc tttgaagata gtaactattt
    14521 ttcggtggtg tttacccggg aaaccgggat gacgcccagc cagtggcgtc atctcaattc
    14581 gcagaaagat taattcgcca tgccc
//
