LOCUS       Exported               14589 bp DNA     circular SYN 22-AUG-2024
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 14589)
  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 14589)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 14589)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 14589)
  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"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..14589
                     /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
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(1105..1327)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(1819..2631)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFL"
     CDS             2979..7082
                     /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(7218..8093)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        8120..8404
                     /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             8431..8439
                     /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             8448..8861
                     /codon_start=1
                     /label=Gam
                     /note="inhibitor of the host RecBCD nuclease in the lambda
                     Red system"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             8870..9652
                     /codon_start=1
                     /label=Beta
                     /note="single-stranded DNA binding recombinase in the
                     lambda Red system"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             9652..10329
                     /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      10333..10577
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             10914..12332
                     /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(12458..12533)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(12555..12673)
                     /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             12819..13652
                     /codon_start=1
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     CDS             13729..14574
                     /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 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 tctgattata 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 tcattggcaa 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 ggataagaaa tactcaatag
     3001 gcttagatat cggcacaaat agcgtcggat gggcggtgat cactgatgat tataaggttc
     3061 cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa aaaaatctta
     3121 taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc aaacggacag
     3181 ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag attttttcaa
     3241 atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct tttttggtgg
     3301 aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat gaagttgctt
     3361 atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat tctactgata
     3421 aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt cgtggtcatt
     3481 ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta tttatccagt
     3541 tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt ggagtagatg
     3601 ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat ctcattgctc
     3661 agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg tcattgggtt
     3721 tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta cagctttcaa
     3781 aagatactta cgatgatgat ttagataatt tattggcgca aattggagat caatatgctg
     3841 atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat atcctaagag
     3901 taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc tacgatgaac
     3961 atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca gaaaagtata
     4021 aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat gggggagcta
     4081 gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat ggtactgagg
     4141 aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc tttgacaacg
     4201 gctctattcc ccatcaaatt cacttgggtg agctgcatgc tattttgaga agacaagaag
     4261 acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg acttttcgaa
     4321 ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg atgactcgga
     4381 agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa ggtgcttcag
     4441 ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat gaaaaagtac
     4501 taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg acaaaggtca
     4561 aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag aagaaagcca
     4621 ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta aaagaagatt
     4681 atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa gatagattta
     4741 atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa gattttttgg
     4801 ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc ttatttgaag
     4861 atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat gataaggtga
     4921 tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga aaattgatta
     4981 atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa tcagatggtt
     5041 ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt aaagaagaca
     5101 ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt gcaaatttag
     5161 ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt gatgaattgg
     5221 tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca cgtgaaaatc
     5281 agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc gaagaaggta
     5341 tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact caattgcaaa
     5401 atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg gaccaagaat
     5461 tagatattaa tcgtttaagt gattatgatg tcgatcacat tgttccacaa agtttcctta
     5521 aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt ggtaaatcgg
     5581 ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga caacttctaa
     5641 acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa cgtggaggtt
     5701 tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact cgccaaatca
     5761 ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat gaaaatgata
     5821 aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct gacttccgaa
     5881 aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc catgatgcgt
     5941 atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt gaatcggagt
     6001 ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag tctgagcaag
     6061 aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac ttcttcaaaa
     6121 cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa actaatgggg
     6181 aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc aaagtattgt
     6241 ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga ttctccaagg
     6301 agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa gactgggatc
     6361 caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta gtggttgcta
     6421 aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta gggatcacaa
     6481 ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct aaaggatata
     6541 aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt gagttagaaa
     6601 acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat gagctggctc
     6661 tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag ttgaagggta
     6721 gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat tatttagatg
     6781 agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat gccaatttag
     6841 ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa caagcagaaa
     6901 atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt aaatattttg
     6961 atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat gccactctta
     7021 tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag ctaggaggtg
     7081 actgaagtat attttagatg aagattattt cttaatctag acatgagcgg atacatattt
     7141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7201 cctgcatcga tttattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
     7261 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaagtagagt
     7321 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
     7381 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
     7441 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     7501 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
     7561 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
     7621 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
     7681 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
     7741 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
     7801 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
     7861 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
     7921 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
     7981 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
     8041 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
     8101 atactcccgc cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac
     8161 tgcgtctttt actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt
     8221 ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac
     8281 ggcagaaaag tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt
     8341 ttatccataa gattagcgga tcctacctga cgctttttat cgcaactctc tactgtttct
     8401 ccatacccgt ttttttggga attcgagctc taaggaggtt ataaaaaatg gatattaata
     8461 ctgaaactga gatcaagcaa aagcattcac taaccccctt tcctgttttc ctaatcagcc
     8521 cggcatttcg cgggcgatat tttcacagct atttcaggag ttcagccatg aacgcttatt
     8581 acattcagga tcgtcttgag gctcagagct gggcgcgtca ctaccagcag ctcgcccgtg
     8641 aagagaaaga ggcagaactg gcagacgaca tggaaaaagg cctgccccag cacctgtttg
     8701 aatcgctatg catcgatcat ttgcaacgcc acggggccag caaaaaatcc attacccgtg
     8761 cgtttgatga cgatgttgag tttcaggagc gcatggcaga acacatccgg tacatggttg
     8821 aaaccattgc tcaccaccag gttgatattg attcagaggt ataaaacgaa tgagtactgc
     8881 actcgcaacg ctggctggga agctggctga acgtgtcggc atggattctg tcgacccaca
     8941 ggaactgatc accactcttc gccagacggc atttaaaggt gatgccagcg atgcgcagtt
     9001 catcgcatta ctgatcgttg ccaaccagta cggccttaat ccgtggacga aagaaattta
     9061 cgcctttcct gataagcaga atggcatcgt tccggtggtg ggcgttgatg gctggtcccg
     9121 catcatcaat gaaaaccagc agtttgatgg catggacttt gagcaggaca atgaatcctg
     9181 tacatgccgg atttaccgca aggaccgtaa tcatccgatc tgcgttaccg aatggatgga
     9241 tgaatgccgc cgcgaaccat tcaaaactcg cgaaggcaga gaaatcacgg ggccgtggca
     9301 gtcgcatccc aaacggatgt tacgtcataa agccatgatt cagtgtgccc gtctggcctt
     9361 cggatttgct ggtatctatg acaaggatga agccgagcgc attgtcgaaa atactgcata
     9421 cactgcagaa cgtcagccgg aacgcgacat cactccggtt aacgatgaaa ccatgcagga
     9481 gattaacact ctgctgatcg ccctggataa aacatgggat gacgacttat tgccgctctg
     9541 ttcccagata tttcgccgcg acattcgtgc atcgtcagaa ctgacacagg ccgaagcagt
     9601 aaaagctctt ggattcctga aacagaaagc cgcagagcag aaggtggcag catgacaccg
     9661 gacattatcc tgcagcgtac cgggatcgat gtgagagctg tcgaacaggg ggatgatgcg
     9721 tggcacaaat tacggctcgg cgtcatcacc gcttcagaag ttcacaacgt gatagcaaaa
     9781 ccccgctccg gaaagaagtg gcctgacatg aaaatgtcct acttccacac cctgcttgct
     9841 gaggtttgca ccggtgtggc tccggaagtt aacgctaaag cactggcctg gggaaaacag
     9901 tacgagaacg acgccagaac cctgtttgaa ttcacttccg gcgtgaatgt tactgaatcc
     9961 ccgatcatct atcgcgacga aagtatgcgt accgcctgct ctcccgatgg tttatgcagt
    10021 gacggcaacg gccttgaact gaaatgcccg tttacctccc gggatttcat gaagttccgg
    10081 ctcggtggtt tcgaggccat aaagtcagct tacatggccc aggtgcagta cagcatgtgg
    10141 gtgacgcgaa aaaatgcctg gtactttgcc aactatgacc cgcgtatgaa gcgtgaaggc
    10201 ctgcattatg tcgtgattga gcgggatgaa aagtacatgg cgagttttga cgagatcgtg
    10261 ccggagttca tcgaaaaaat ggacgaggca ctggctgaaa ttggttttgt atttggggag
    10321 caatggcgat gacgcatcct cacgataata tccgggtagg cgcaatcact ttcgtctact
    10381 ccgttacaaa gcgaggctgg gtatttcccg gcctttctgt tatccgaaat ccactgaaag
    10441 cacagcggct ggctgaggag ataaataata aacgaggggc tgtatgcaca aagcatcttc
    10501 tgttgagtta agaacgagta tcgagatggc acatagcctt gctcaaattg gaatcaggtt
    10561 tgtgccaata ccagtagaaa cagacgaaga atccatgggt atggacagat cctctttagg
    10621 cccgtagtct gcaaatcctt ttatgatttt ctatcaaaca aaagaggaaa atagaccagt
    10681 tgcaatccaa acgagagtct aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta
    10741 ctgataaagc aggcaagacc taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc
    10801 ccttacatat tttaggtctt tttttattgt gcgtaactaa cttgccatct tcaaacagga
    10861 gggctggaag aagcagaccg ctaacacagt acataaaaaa ggagacatga acgatgaaca
    10921 tcaaaaagtt tgcaaaacaa gcaacagtat taacctttac taccgcactg ctggcaggag
    10981 gcgcaactca agcgtttgcg aaagaaacga accaaaagcc atataaggaa acatacggca
    11041 tttcccatat tacacgccat gatatgctgc aaatccctga acagcaaaaa aatgaaaaat
    11101 atcaagttcc tgaattcgat tcgtccacaa ttaaaaatat ctcttctgca aaaggcctgg
    11161 acgtttggga cagctggcca ttacaaaacg ctgacggcac tgtcgcaaac tatcacggct
    11221 accacatcgt ctttgcatta gccggagatc ctaaaaatgc ggatgacaca tcgatttaca
    11281 tgttctatca aaaagtcggc gaaacttcta ttgacagctg gaaaaacgct ggccgcgtct
    11341 ttaaagacag cgacaaattc gatgcaaatg attctatcct aaaagaccaa acacaagaat
    11401 ggtcaggttc agccacattt acatctgacg gaaaaatccg tttattctac actgatttct
    11461 ccggtaaaca ttacggcaaa caaacactga caactgcaca agttaacgta tcagcatcag
    11521 acagctcttt gaacatcaac ggtgtagagg attataaatc aatctttgac ggtgacggaa
    11581 aaacgtatca aaatgtacag cagttcatcg atgaaggcaa ctacagctca ggcgacaacc
    11641 atacgctgag agatcctcac tacgtagaag ataaaggcca caaatactta gtatttgaag
    11701 caaacactgg aactgaagat ggctaccaag gcgaagaatc tttatttaac aaagcatact
    11761 atggcaaaag cacatcattc ttccgtcaag aaagtcaaaa acttctgcaa agcgataaaa
    11821 aacgcacggc tgagttagca aacggcgctc tcggtatgat tgagctaaac gatgattaca
    11881 cactgaaaaa agtgatgaaa ccgctgattg catctaacac agtaacagat gaaattgaac
    11941 gcgcgaacgt ctttaaaatg aacggcaaat ggtacctgtt cactgactcc cgcggatcaa
    12001 aaatgacgat tgacggcatt acgtctaacg atatttacat gcttggttat gtttctaatt
    12061 ctttaactgg cccatacaag ccgctgaaca aaactggcct tgtgttaaaa atggatcttg
    12121 atcctaacga tgtaaccttt acttactcac acttcgctgt acctcaagcg aaaggaaaca
    12181 atgtcgtgat tacaagctat atgacaaaca gaggattcta cgcagacaaa caatcaacgt
    12241 ttgcgccaag cttcctgctg aacatcaaag gcaagaaaac atctgttgtc aaagacagca
    12301 tccttgaaca aggacaatta acagttaaca aataaaaacg caaaagaaaa tgccgatatc
    12361 ctattggcat tttcttttat ttcttatcaa cataaaggtg aatcccatat gaactatata
    12421 aaagcaggca aatggctaac cgtattcctc aaaaaaagca ccgactcggt gccacttttt
    12481 caagttgata acggactagc cttattttaa cttgctattt ctagctctaa aacggttgga
    12541 ctcaagacga tagttacgac cagtctaaaa agcgcctgaa ttcgcgacct tctcgttact
    12601 gacaggaaaa tgggccattg gcaaccaggg aaagatgaac gtgatgatgt tcacaatttg
    12661 ctgaattgtg gtgatgtgat gctcaccgca tttcctgaaa attcacgctg tatcttgaaa
    12721 aatcgacgtt ttttacgtgg ttttccgtcg aaaatttaag gtaagaacct gacctcgtga
    12781 ttactatttc gccgtgttga cgacatcagg aggccagtat gaccgtatta catagtgtgg
    12841 atttttttcc gtctggtaac gcgtccgtgg cgatagaacc ccggctcccg caggcggatt
    12901 ttcctgaaca tcatcatgat tttcatgaaa ttgtgattgt cgaacatggc acgggtattc
    12961 atgtgtttaa tgggcagccc tataccatca ccggtggcac ggtctgtttc gtacgcgatc
    13021 atgatcggca tctgtatgaa cataccgata atctgtgtct gaccaatgtg ctgtatcgct
    13081 cgccggatcg atttcagttt ctcgccgggc tgaatcagtt gctgccacaa gagctggatg
    13141 ggcagtatcc gtctcactgg cgcgttaacc acagcgtatt gcagcaggtg cgacagctgg
    13201 ttgcacagat ggaacagcag gaaggggaaa atgatttacc ctcgaccgcc agtcgcgaga
    13261 tcttgtttat gcaattactg ctcttgctgc gtaaaagcag tttgcaggag aacctggaaa
    13321 acagcgcatc acgtctcaac ttgcttctgg cctggctgga ggaccatttt gccgatgagg
    13381 tgaattggga tgccgtggcg gatcaatttt ctctttcact gcgtacgcta catcggcagc
    13441 ttaagcagca aacgggactg acgcctcagc gatacctgaa ccgcctgcga ctgatgaaag
    13501 cccgacatct gctacgccac agcgaggcca gcgttactga catcgcctat cgctgtggat
    13561 tcagcgacag taaccacttt tcgacgcttt ttcgccgaga gtttaactgg tcaccgcgtg
    13621 atattcgcca gggacgggat ggctttctgc aataacgcga atcttctcaa cgtatttgta
    13681 cgccatattg cgaataatca acttcgttct ctggccgagg tagccacggt ggcgcatcag
    13741 ttaaaacttc tcaaagatga tttttttgcc agcgaccagc aggcagtcgc tgtggctgac
    13801 cgttatccgc aagatgtctt tgctgaacat acacatgatt tttgtgagct ggtgattgtc
    13861 tggcgcggta atggcctgca tgtactcaac gatcgccctt atcgcattac ccgtggcgat
    13921 ctcttttaca ttcatgctga cgataaacac tcctacgctt ccgttaacga tctggttttg
    13981 cagaatatta tttattgccc ggagcgtctg aagctgaatc ttgactggca gggggcgatt
    14041 ccgggattta acgccagcgc agggcaacca cactggcgct taggtagcat ggggatggcg
    14101 caggcgcggc aggttatcgg tcagcttgag catgaaagta gtcagcatgt gccgtttgct
    14161 aacgaaatgg ctgagttgct gttcgggcag ttggtgatgt tgctgaatcg ccatcgttac
    14221 accagtgatt cgttgccgcc aacatccagc gaaacgttgc tggataagct gattacccgg
    14281 ctggcggcta gcctgaaaag tccctttgcg ctggataaat tttgtgatga ggcatcgtgc
    14341 agtgagcgcg ttttgcgtca gcaatttcgc cagcagactg gaatgaccat caatcaatat
    14401 ctgcgacagg tcagagtgtg tcatgcgcaa tatcttctcc agcatagccg cctgttaatc
    14461 agtgatattt cgaccgaatg tggctttgaa gatagtaact atttttcggt ggtgtttacc
    14521 cgggaaaccg ggatgacgcc cagccagtgg cgtcatctca attcgcagaa agattaattc
    14581 gccatgccc
//
