LOCUS       sequence-458133-       11918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ISEre1 controled by pCas promoter, lambda-red 
            recombinase, SacB and ISFba1 guide RNA targeting pMB1 ori.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11918)
  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 11918)
  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: pISFba1
FEATURES             Location/Qualifiers
     source          1..11918
                     /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(4272..5150)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5177..5461
                     /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     5384..5403
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             5488..5496
                     /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             5505..5921
                     /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             5927..6712
                     /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             6709..7389
                     /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
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      7390..7634
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             join(7971..8057,8058..9392)
                     /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(9509..9556)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     9552..9570
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     ncRNA           complement(9609..9676)
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     promoter        complement(9884..10002)
                     /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             10148..10984
                     /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             11058..11906
                     /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 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 gtggaaattg ctaaagagct
     3001 tcaagacgga aataaatccg tcagaggaac agaaagtcaa gattcataag acgataggaa
     3061 cttgtagatt catttataat ttctatcttg ctcataacaa agaactttac gataaaggcg
     3121 aaaagtttat gagtggtaaa agttttagtg tttggctgaa caatgagtat cttccacaaa
     3181 atccagacaa gttatggatt aaggaagtca gttcaaaatc tgtaaagcac tcaattgaaa
     3241 atggttgtat cgcatttacc agatttttca aacatcaaag tgcttttccg aatttaaaaa
     3301 agaaaggcaa atctgatgta aaaatgtatt ttgtaaagaa taatccaaaa gattgtcgtt
     3361 gcgaaagaca cagaatcaat attccgtcac ttggttgggt tcgcatcaaa gaaaaaggat
     3421 atattccaac aactaaagac ggatatgtga ttaaaagcgg cacggtctca atgaaagctg
     3481 acagatacta tgtttcagtt cttgtagaaa tttcaaataa caagatagcc aataattcca
     3541 atgcaggaat tggcattgac cttggattaa aggattttgc cattgtttct aatggtaaaa
     3601 cttataagaa cattaacaaa tctgcaagat taaagaaaca tgaaaaacag cttattcgag
     3661 aacagagaag tctttctcga aagtatgaaa atttaaagaa aggagagtcc actcaaaaag
     3721 caaatataca aaaacaaagg cttaaggtac agaaacttca tcatagaatg gataatattc
     3781 gtactgatta catcaataaa acaatcgcag agatagtaaa aaccaaacca tcttatatta
     3841 cgattgaaga tttgaatgta aaaggtatga tgaaaaacag acatctttca aaagctgttg
     3901 cttcacagaa gttttatgaa tttagaacaa agcttcaagc taaatgcaat gaaaatggaa
     3961 tcgaacttcg agtagtagac agatggtatc catcatctaa aacctgtcac tgctgtggcg
     4021 ctgtcaaaaa agatttaaag ctttcagata gaatatttaa atgcagttgc ggttatgtcg
     4081 aagacagaga tttcaatgct gcacttaatt tgagagatgc gataacttac gaagttgcat
     4141 gaagtatatt ttagatgaag attatttctt aatctagaca tgagcggata catatttgaa
     4201 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4261 gcatcgattt attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca
     4321 cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa gtagagttga
     4381 tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc
     4441 agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc
     4501 tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg
     4561 atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga
     4621 ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc
     4681 tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg
     4741 atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc
     4801 gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag
     4861 taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct
     4921 cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc
     4981 accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg
     5041 tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg
     5101 gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata
     5161 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
     5221 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
     5281 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
     5341 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
     5401 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca
     5461 tacccgtttt tttgggaatt cgagctctaa ggaggttata aaaaatggat attaatactg
     5521 aaactgagat caagcaaaag cattcactaa ccccctttcc tgttttccta atcagcccgg
     5581 catttcgcgg gcgatatttt cacagctatt tcaggagttc agccatgaac gcttattaca
     5641 ttcaggatcg tcttgaggct cagagctggg cgcgtcacta ccagcagctc gcccgtgaag
     5701 agaaagaggc agaactggca gacgacatgg aaaaaggcct gccccagcac ctgtttgaat
     5761 cgctatgcat cgatcatttg caacgccacg gggccagcaa aaaatccatt acccgtgcgt
     5821 ttgatgacga tgttgagttt caggagcgca tggcagaaca catccggtac atggttgaaa
     5881 ccattgctca ccaccaggtt gatattgatt cagaggtata aaacgaatga gtactgcact
     5941 cgcaacgctg gctgggaagc tggctgaacg tgtcggcatg gattctgtcg acccacagga
     6001 actgatcacc actcttcgcc agacggcatt taaaggtgat gccagcgatg cgcagttcat
     6061 cgcattactg atcgttgcca accagtacgg ccttaatccg tggacgaaag aaatttacgc
     6121 ctttcctgat aagcagaatg gcatcgttcc ggtggtgggc gttgatggct ggtcccgcat
     6181 catcaatgaa aaccagcagt ttgatggcat ggactttgag caggacaatg aatcctgtac
     6241 atgccggatt taccgcaagg accgtaatca tccgatctgc gttaccgaat ggatggatga
     6301 atgccgccgc gaaccattca aaactcgcga aggcagagaa atcacggggc cgtggcagtc
     6361 gcatcccaaa cggatgttac gtcataaagc catgattcag tgtgcccgtc tggccttcgg
     6421 atttgctggt atctatgaca aggatgaagc cgagcgcatt gtcgaaaata ctgcatacac
     6481 tgcagaacgt cagccggaac gcgacatcac tccggttaac gatgaaacca tgcaggagat
     6541 taacactctg ctgatcgccc tggataaaac atgggatgac gacttattgc cgctctgttc
     6601 ccagatattt cgccgcgaca ttcgtgcatc gtcagaactg acacaggccg aagcagtaaa
     6661 agctcttgga ttcctgaaac agaaagccgc agagcagaag gtggcagcat gacaccggac
     6721 attatcctgc agcgtaccgg gatcgatgtg agagctgtcg aacaggggga tgatgcgtgg
     6781 cacaaattac ggctcggcgt catcaccgct tcagaagttc acaacgtgat agcaaaaccc
     6841 cgctccggaa agaagtggcc tgacatgaaa atgtcctact tccacaccct gcttgctgag
     6901 gtttgcaccg gtgtggctcc ggaagttaac gctaaagcac tggcctgggg aaaacagtac
     6961 gagaacgacg ccagaaccct gtttgaattc acttccggcg tgaatgttac tgaatccccg
     7021 atcatctatc gcgacgaaag tatgcgtacc gcctgctctc ccgatggttt atgcagtgac
     7081 ggcaacggcc ttgaactgaa atgcccgttt acctcccggg atttcatgaa gttccggctc
     7141 ggtggtttcg aggccataaa gtcagcttac atggcccagg tgcagtacag catgtgggtg
     7201 acgcgaaaaa atgcctggta ctttgccaac tatgacccgc gtatgaagcg tgaaggcctg
     7261 cattatgtcg tgattgagcg ggatgaaaag tacatggcga gttttgacga gatcgtgccg
     7321 gagttcatcg aaaaaatgga cgaggcactg gctgaaattg gttttgtatt tggggagcaa
     7381 tggcgatgac gcatcctcac gataatatcc gggtaggcgc aatcactttc gtctactccg
     7441 ttacaaagcg aggctgggta tttcccggcc tttctgttat ccgaaatcca ctgaaagcac
     7501 agcggctggc tgaggagata aataataaac gaggggctgt atgcacaaag catcttctgt
     7561 tgagttaaga acgagtatcg agatggcaca tagccttgct caaattggaa tcaggtttgt
     7621 gccaatacca gtagaaacag acgaagaatc catgggtatg gacagatcct ctttaggccc
     7681 gtagtctgca aatcctttta tgattttcta tcaaacaaaa gaggaaaata gaccagttgc
     7741 aatccaaacg agagtctaat agaatgaggt cgaaaagtaa atcgcgcggg tttgttactg
     7801 ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg tatactttgg cgtcacccct
     7861 tacatatttt aggtcttttt ttattgtgcg taactaactt gccatcttca aacaggaggg
     7921 ctggaagaag cagaccgcta acacagtaca taaaaaagga gacatgaacg atgaacatca
     7981 aaaagtttgc aaaacaagca acagtattaa cctttactac cgcactgctg gcaggaggcg
     8041 caactcaagc gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca tacggcattt
     8101 cccatattac acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat gaaaaatatc
     8161 aagttcctga attcgattcg tccacaatta aaaatatctc ttctgcaaaa ggcctggacg
     8221 tttgggacag ctggccatta caaaacgctg acggcactgt cgcaaactat cacggctacc
     8281 acatcgtctt tgcattagcc ggagatccta aaaatgcgga tgacacatcg atttacatgt
     8341 tctatcaaaa agtcggcgaa acttctattg acagctggaa aaacgctggc cgcgtcttta
     8401 aagacagcga caaattcgat gcaaatgatt ctatcctaaa agaccaaaca caagaatggt
     8461 caggttcagc cacatttaca tctgacggaa aaatccgttt attctacact gatttctccg
     8521 gtaaacatta cggcaaacaa acactgacaa ctgcacaagt taacgtatca gcatcagaca
     8581 gctctttgaa catcaacggt gtagaggatt ataaatcaat ctttgacggt gacggaaaaa
     8641 cgtatcaaaa tgtacagcag ttcatcgatg aaggcaacta cagctcaggc gacaaccata
     8701 cgctgagaga tcctcactac gtagaagata aaggccacaa atacttagta tttgaagcaa
     8761 acactggaac tgaagatggc taccaaggcg aagaatcttt atttaacaaa gcatactatg
     8821 gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc gataaaaaac
     8881 gcacggctga gttagcaaac ggcgctctcg gtatgattga gctaaacgat gattacacac
     8941 tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt aacagatgaa attgaacgcg
     9001 cgaacgtctt taaaatgaac ggcaaatggt acctgttcac tgactcccgc ggatcaaaaa
     9061 tgacgattga cggcattacg tctaacgata tttacatgct tggttatgtt tctaattctt
     9121 taactggccc atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg gatcttgatc
     9181 ctaacgatgt aacctttact tactcacact tcgctgtacc tcaagcgaaa ggaaacaatg
     9241 tcgtgattac aagctatatg acaaacagag gattctacgc agacaaacaa tcaacgtttg
     9301 cgccaagctt cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa gacagcatcc
     9361 ttgaacaagg acaattaaca gttaacaaat aaaaacgcaa aagaaaatgc cgatatccta
     9421 ttggcatttt cttttatttc ttatcaacat aaaggtgaat cccatatgaa ctatataaaa
     9481 gcaggcaaat ggctaaccgt attcctcaca aaaaacccct caagacccgt ttagaggccc
     9541 caaggggtta tgctagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg
     9601 ctttgttagt cccattcgcc atgccgaagc atgttgccca gccggcgcca gcgaggaggc
     9661 tgggaccatg ccggcctgga agctccctcg tgcgctctgc cactcaaaat gtgattaaat
     9721 atgctcacat cgagataatg taagttctca cgaacattac ttactatttc atcgtgtatg
     9781 ctttggattg accgaaatca tatctactca caagttcttg tacactccat agtcgtaaat
     9841 tcccgaaaat agccttcggt acatacttac gtttgcgttt tagtacgacc agtctaaaaa
     9901 gcgcctgaat tcgcgacctt ctcgttactg acaggaaaat gggccattgg caaccaggga
     9961 aagatgaacg tgatgatgtt cacaatttgc tgaattgtgg tgatgtgatg ctcaccgcat
    10021 ttcctgaaaa ttcacgctgt atcttgaaaa atcgacgttt tttacgtggt tttccgtcga
    10081 aaatttaagg taagaacctg acctcgtgat tactatttcg ccgtgttgac gacatcagga
    10141 ggccagtatg accgtattac atagtgtgga tttttttccg tctggtaacg cgtccgtggc
    10201 gatagaaccc cggctcccgc aggcggattt tcctgaacat catcatgatt ttcatgaaat
    10261 tgtgattgtc gaacatggca cgggtattca tgtgtttaat gggcagccct ataccatcac
    10321 cggtggcacg gtctgtttcg tacgcgatca tgatcggcat ctgtatgaac ataccgataa
    10381 tctgtgtctg accaatgtgc tgtatcgctc gccggatcga tttcagtttc tcgccgggct
    10441 gaatcagttg ctgccacaag agctggatgg gcagtatccg tctcactggc gcgttaacca
    10501 cagcgtattg cagcaggtgc gacagctggt tgcacagatg gaacagcagg aaggggaaaa
    10561 tgatttaccc tcgaccgcca gtcgcgagat cttgtttatg caattactgc tcttgctgcg
    10621 taaaagcagt ttgcaggaga acctggaaaa cagcgcatca cgtctcaact tgcttctggc
    10681 ctggctggag gaccattttg ccgatgaggt gaattgggat gccgtggcgg atcaattttc
    10741 tctttcactg cgtacgctac atcggcagct taagcagcaa acgggactga cgcctcagcg
    10801 atacctgaac cgcctgcgac tgatgaaagc ccgacatctg ctacgccaca gcgaggccag
    10861 cgttactgac atcgcctatc gctgtggatt cagcgacagt aaccactttt cgacgctttt
    10921 tcgccgagag tttaactggt caccgcgtga tattcgccag ggacgggatg gctttctgca
    10981 ataacgcgaa tcttctcaac gtatttgtac gccatattgc gaataatcaa cttcgttctc
    11041 tggccgaggt agccacggtg gcgcatcagt taaaacttct caaagatgat ttttttgcca
    11101 gcgaccagca ggcagtcgct gtggctgacc gttatccgca agatgtcttt gctgaacata
    11161 cacatgattt ttgtgagctg gtgattgtct ggcgcggtaa tggcctgcat gtactcaacg
    11221 atcgccctta tcgcattacc cgtggcgatc tcttttacat tcatgctgac gataaacact
    11281 cctacgcttc cgttaacgat ctggttttgc agaatattat ttattgcccg gagcgtctga
    11341 agctgaatct tgactggcag ggggcgattc cgggatttaa cgccagcgca gggcaaccac
    11401 actggcgctt aggtagcatg gggatggcgc aggcgcggca ggttatcggt cagcttgagc
    11461 atgaaagtag tcagcatgtg ccgtttgcta acgaaatggc tgagttgctg ttcgggcagt
    11521 tggtgatgtt gctgaatcgc catcgttaca ccagtgattc gttgccgcca acatccagcg
    11581 aaacgttgct ggataagctg attacccggc tggcggctag cctgaaaagt ccctttgcgc
    11641 tggataaatt ttgtgatgag gcatcgtgca gtgagcgcgt tttgcgtcag caatttcgcc
    11701 agcagactgg aatgaccatc aatcaatatc tgcgacaggt cagagtgtgt catgcgcaat
    11761 atcttctcca gcatagccgc ctgttaatca gtgatatttc gaccgaatgt ggctttgaag
    11821 atagtaacta tttttcggtg gtgtttaccc gggaaaccgg gatgacgccc agccagtggc
    11881 gtcatctcaa ttcgcagaaa gattaattcg ccatgccc
//
