LOCUS       sequence-482811-       11825 bp    DNA     circular SYN 05-SEP-2025
DEFINITION  Expresses lambda red machinery and cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11825)
  TITLE     Iterative in vivo cut'n'paste of chromosomal loci in Escherichia 
            coli K-12 using synthetic DNA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11825)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pRedCas9
FEATURES             Location/Qualifiers
     source          1..11825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(122..366)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(367..1047)
                     /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"
     CDS             complement(1044..1829)
                     /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             complement(1835..2251)
                     /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"
     RBS             2260..2268
                     /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)"
     promoter        complement(2295..2579)
                     /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     complement(2353..2372)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2606..3484
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      complement(3535..3566)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     misc_RNA        4807..4856
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             4903..9006
                     /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             4903..9006
                     /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             9016..9039
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        9087..9190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9191..9982
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      10604..10826
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             10874..11824
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 gttcttatgg ctcttgtatc tatcagtgaa gcatcaagac taacaaacaa aagtagaaca
       61 actgttcacc gttacatatc aaagggaaaa ctgtccatac ccatggattc ttcgtctgtt
      121 tctactggta ttggcacaaa cctgattcca atttgagcaa ggctatgtgc catctcgata
      181 ctcgttctta actcaacaga agatgctttg tgcatacagc ccctcgttta ttatttatct
      241 cctcagccag ccgctgtgct ttcagtggat ttcggataac agaaaggccg ggaaataccc
      301 agcctcgctt tgtaacggag tagacgaaag tgattgcgcc tacccggata ttatcgtgag
      361 gatgcgtcat cgccattgct ccccaaatac aaaaccaatt tcagccagtg cctcgtccat
      421 tttttcgatg aactccggca cgatctcgtc aaaactcgcc atgtactttt catcccgctc
      481 aatcacgaca taatgcaggc cttcacgctt catacgcggg tcatagttgg caaagtacca
      541 ggcatttttt cgcgtcaccc acatgctgta ctgcacctgg gccatgtaag ctgactttat
      601 ggcctcgaaa ccaccgagcc ggaacttcat gaaatcccgg gaggtaaacg ggcatttcag
      661 ttcaaggccg ttgccgtcac tgcataaacc atcgggagag caggcggtac gcatactttc
      721 gtcgcgatag atgatcgggg attcagtaac attcacgccg gaagtgaatt caaacagggt
      781 tctggcgtcg ttctcgtact gttttcccca ggccagtgct ttagcgttaa cttccggagc
      841 cacaccggtg caaacctcag caagcagggt gtggaagtag gacattttca tgtcaggcca
      901 cttctttccg gagcggggtt ttgctatcac gttgtgaact tctgaagcgg tgatgacgcc
      961 gagccgtaat ttgtgccacg catcatcccc ctgttcgaca gctctcacat cgatcccggt
     1021 acgctgcagg ataatgtccg gtgtcatgct gccaccttct gctctgcggc tttctgtttc
     1081 aggaatccaa gagcttttac tgcttcggcc tgtgtcagtt ctgacgatgc acgaatgtcg
     1141 cggcgaaata tctgggaaca gagcggcaat aagtcgtcat cccatgtttt atccagggcg
     1201 atcagcagag tgttaatctc ctgcatggtt tcatcgttaa ccggagtgat gtcgcgttcc
     1261 ggctgacgtt ctgcagtgta tgcagtattt tcgacaatgc gctcggcttc atccttgtca
     1321 tagataccag caaatccgaa ggccagacgg gcacactgaa tcatggcttt atgacgtaac
     1381 atccgtttgg gatgcgactg ccacggcccc gtgatttctc tgccttcgcg agttttgaat
     1441 ggttcgcggc ggcattcatc catccattcg gtaacgcaga tcggatgatt acggtccttg
     1501 cggtaaatcc ggcatgtaca ggattcattg tcctgctcaa agtccatgcc atcaaactgc
     1561 tggttttcat tgatgatgcg ggaccagcca tcaacgccca ccaccggaac gatgccattc
     1621 tgcttatcag gaaaggcgta aatttctttc gtccacggat taaggccgta ctggttggca
     1681 acgatcagta atgcgatgaa ctgcgcatcg ctggcatcac ctttaaatgc cgtctggcga
     1741 agagtggtga tcagttcctg tgggtcgaca gaatccatgc cgacacgttc agccagcttc
     1801 ccagccagcg ttgcgagtgc agtactcatt cgttttatac ctctgaatca atatcaacct
     1861 ggtggtgagc aatggtttca accatgtacc ggatgtgttc tgccatgcgc tcctgaaact
     1921 caacatcgtc atcaaacgca cgggtaatgg attttttgct ggccccgtgg cgttgcaaat
     1981 gatcgatgca tagcgattca aacaggtgct ggggcaggcc tttttccatg tcgtctgcca
     2041 gttctgcctc tttctcttca cgggcgagct gctggtagtg acgcgcccag ctctgagcct
     2101 caagacgatc ctgaatgtaa taagcgttca tggctgaact cctgaaatag ctgtgaaaat
     2161 atcgcccgcg aaatgccggg ctgattagga aaacaggaaa gggggttagt gaatgctttt
     2221 gcttgatctc agtttcagta ttaatatcca ttttttataa cctccttaga gctcgaattc
     2281 ccaaaaaaac gggtatggag aaacagtaga gagttgcgat aaaaagcgtc aggtaggatc
     2341 cgctaatctt atggataaaa atgctatggc atagcaaagt gtgacgccgt gcaaataatc
     2401 aatgtggact tttctgccgt gattatagac acttttgtta cgcgtttttg tcatggcttt
     2461 ggtcccgctt tgttacagaa tgcttttaat aagcggggtt accggtttgg ttagcgagaa
     2521 gagccagtaa aagacgcagt gacggcaatg tctgatgcaa tatggacaat tggtttcttc
     2581 tctgaatggc gggagtatga aaagtatggc tgaagcgcaa aatgatcccc tgctgccggg
     2641 atactcgttt aatgcccatc tggtggcggg tttaacgccg attgaggcca acggttatct
     2701 cgattttttt atcgaccgac cgctgggaat gaaaggttat attctcaatc tcaccattcg
     2761 cggtcagggg gtggtgaaaa atcagggacg agaatttgtt tgccgaccgg gtgatatttt
     2821 gctgttcccg ccaggagaga ttcatcacta cggtcgtcat ccggaggctc gcgaatggta
     2881 tcaccagtgg gtttactttc gtccgcgcgc ctactggcat gaatggctta actggccgtc
     2941 aatatttgcc aatacggggt tctttcgccc ggatgaagcg caccagccgc atttcagcga
     3001 cctgtttggg caaatcatta acgccgggca aggggaaggg cgctattcgg agctgctggc
     3061 gataaatctg cttgagcaat tgttactgcg gcgcatggaa gcgattaacg agtcgctcca
     3121 tccaccgatg gataatcggg tacgcgaggc ttgtcagtac atcagcgatc acctggcaga
     3181 cagcaatttt gatatcgcca gcgtcgcaca gcatgtttgc ttgtcgccgt cgcgtctgtc
     3241 acatcttttc cgccagcagt tagggattag cgtcttaagc tggcgcgagg accaacgtat
     3301 cagccaggcg aagctgcttt tgagcaccac ccggatgcct atcgccaccg tcggtcgcaa
     3361 tgttggtttt gacgatcaac tctacttctc gcgggtattt aaaaaatgca ccggggccag
     3421 cccgagcgag ttccgtgccg gttgtgaaga aaaagtgaat gatgtagccg tcaagttgtc
     3481 ataataaatc gatgcaggtg gcacttttcg gggaaatgtg ccggatcccc ctcaagtcaa
     3541 aagcctccgg tcggaggttt ttgactttct gctatggagg tcaggtatga tttaaatggt
     3601 cagtattgag cgatatctag agaattcgtc aacgaattca agcttaaggt tgaaaatttg
     3661 ccaattagga tccttatcaa tccgtaaaca ggtcaaacat cagttgccgc aaccaaatat
     3721 tggctaggtc cttgtggtac ttcgcatgcc agaacatgtt gatggctatt tcaggcaaga
     3781 cgactgggtg cggcaaggcg cttaggccga agggctccac gcagcagtcg gctaaacata
     3841 tcggcacagt ggcgagcaga tcggtgcgct ggaggatgtg gccaacggcg gcgaagtgcg
     3901 gcacttccag acggatgtcg cgccggatgc cgacccgtgt catgtacgtg tccacctcgc
     3961 cgtggccggt gccagcggcg atgacacgca cgtggccgta ggaacagaag cgctccagag
     4021 tcaggggttc gcgggtgact ggatggtcct tgcgacatag gcacacgtag tgattctgga
     4081 gcagccggcg ctgaaagaag ccagtttgca gattgggaag caggcccacg gccaagtcca
     4141 cggttccgtt ctgcaaggcc tgcatcaggc tcatcgaact gtcgcgcacc gtactgatca
     4201 cgcaattggg ggcctggtga gccagcgcat ccatcagccg cggcatgaag tagatctcgc
     4261 caatgtcggt catggccagg gtgaaggtac gctcgctggt cagcggatcg aagctttcat
     4321 ggtgctgtag ggcgttgcgc agtgcgtgca tggccgaagt gacgtgctcg gccagatgcg
     4381 cggcataggg tgtgggttcc attccctgat gtgtgcgcac gaagagtggg tcctgtagcg
     4441 aggtgcgcag gcgtttcagc gcattgctca cggctggctg ggtcaggccc aggttctccg
     4501 cagtgacaga gacgcgtctg tcgaccagca actggttgaa caccaccagc aggtttaaat
     4561 ccaggtcacg cagttccata ttcaaccccc atgaattgac tctcttccgg gcgctatcat
     4621 gccataccgc gaaaggtttt gcaccattcg atggcgcgcc gccattggag cttaacgatc
     4681 gttggctggg ggcctcgctt gggttattgc tggtgcccgg ccgggcgcaa tattcatgtt
     4741 gatgatttat tatatatcga gtggtgtatt tatttatatt gtttgctccg ttaccgttat
     4801 taacagctgt caccggatgt gctttccggt ctgatgagtc cgtgaggacg aaacagcctc
     4861 tacaaataat tttgtttaat actagagaag gaggtacaat caatggataa gaaatactca
     4921 ataggcttag atatcggcac aaatagcgtc ggatgggcgg tgatcactga tgaatataag
     4981 gttccgtcta aaaagttcaa ggttctggga aatacagacc gccacagtat caaaaaaaat
     5041 cttatagggg ctcttttatt tgacagtgga gagacagcgg aagcgactcg tctcaaacgg
     5101 acagctcgta gaaggtatac acgtcggaag aatcgtattt gttatctaca ggagattttt
     5161 tcaaatgaga tggcgaaagt agatgatagt ttctttcatc gacttgaaga gtcttttttg
     5221 gtggaagaag acaagaagca tgaacgtcat cctatttttg gaaatatagt agatgaagtt
     5281 gcttatcatg agaaatatcc aactatctat catctgcgaa aaaaattggt agattctact
     5341 gataaagcgg atttgcgctt aatctatttg gccttagcgc atatgattaa gtttcgtggt
     5401 cattttttga ttgagggaga tttaaatcct gataatagtg atgtggacaa actatttatc
     5461 cagttggtac aaacctacaa tcaattattt gaagaaaacc ctattaacgc aagtggagta
     5521 gatgctaaag cgattctttc tgcacgattg agtaaatcaa gacgattaga aaatctcatt
     5581 gctcagctcc ccggtgagaa gaaaaatggc ttatttggga atctcattgc tttgtcattg
     5641 ggtttgaccc ctaattttaa atcaaatttt gatttggcag aagatgctaa attacagctt
     5701 tcaaaagata cttacgatga tgatttagat aatttattgg cgcaaattgg agatcaatat
     5761 gctgatttgt ttttggcagc taagaattta tcagatgcta ttttactttc agatatccta
     5821 agagtaaata ctgaaataac taaggctccc ctatcagctt caatgattaa acgctacgat
     5881 gaacatcatc aagacttgac tcttttaaaa gctttagttc gacaacaact tccagaaaag
     5941 tataaagaaa tcttttttga tcaatcaaaa aacggatatg caggttatat tgatggggga
     6001 gctagccaag aagaatttta taaatttatc aaaccaattt tagaaaaaat ggatggtact
     6061 gaggaattat tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac
     6121 aacggctcta ttccccatca aattcacttg ggtgagctgc atgctatttt gagaagacaa
     6181 gaagactttt atccattttt aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt
     6241 cgaattcctt attatgttgg tccattggcg cgtggcaata gtcgttttgc atggatgact
     6301 cggaagtctg aagaaacaat taccccatgg aattttgaag aagttgtcga taaaggtgct
     6361 tcagctcaat catttattga acgcatgaca aactttgata aaaatcttcc aaatgaaaaa
     6421 gtactaccaa aacatagttt gctttatgag tattttacgg tttataacga attgacaaag
     6481 gtcaaatatg ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa
     6541 gccattgttg atttactctt caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa
     6601 gattatttca aaaaaataga atgttttgat agtgttgaaa tttcaggagt tgaagataga
     6661 tttaatgctt cattaggtac ctaccatgat ttgctaaaaa ttattaaaga taaagatttt
     6721 ttggataatg aagaaaatga agatatctta gaggatattg ttttaacatt gaccttattt
     6781 gaagataggg agatgattga ggaaagactt aaaacatatg ctcacctctt tgatgataag
     6841 gtgatgaaac agcttaaacg tcgccgttat actggttggg gacgtttgtc tcgaaaattg
     6901 attaatggta ttagggataa gcaatctggc aaaacaatat tagatttttt gaaatcagat
     6961 ggttttgcca atcgcaattt tatgcagctg atccatgatg atagtttgac atttaaagaa
     7021 gacattcaaa aagcacaagt gtctggacaa ggcgatagtt tacatgaaca tattgcaaat
     7081 ttagctggta gccctgctat taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa
     7141 ttggtcaaag taatggggcg gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa
     7201 aatcagacaa ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa
     7261 ggtatcaaag aattaggaag tcagattctt aaagagcatc ctgttgaaaa tactcaattg
     7321 caaaatgaaa agctctatct ctattatctc caaaatggaa gagacatgta tgtggaccaa
     7381 gaattagata ttaatcgttt aagtgattat gatgtcgatc acattgttcc acaaagtttc
     7441 cttaaagacg attcaataga caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa
     7501 tcggataacg ttccaagtga agaagtagtc aaaaagatga aaaactattg gagacaactt
     7561 ctaaacgcca agttaatcac tcaacgtaag tttgataatt taacgaaagc tgaacgtgga
     7621 ggtttgagtg aacttgataa agctggtttt atcaaacgcc aattggttga aactcgccaa
     7681 atcactaagc atgtggcaca aattttggat agtcgcatga atactaaata cgatgaaaat
     7741 gataaactta ttcgagaggt taaagtgatt accttaaaat ctaaattagt ttctgacttc
     7801 cgaaaagatt tccaattcta taaagtacgt gagattaaca attaccatca tgcccatgat
     7861 gcgtatctaa atgccgtcgt tggaactgct ttgattaaga aatatccaaa acttgaatcg
     7921 gagtttgtct atggtgatta taaagtttat gatgttcgta aaatgattgc taagtctgag
     7981 caagaaatag gcaaagcaac cgcaaaatat ttcttttact ctaatatcat gaacttcttc
     8041 aaaacagaaa ttacacttgc aaatggagag attcgcaaac gccctctaat cgaaactaat
     8101 ggggaaactg gagaaattgt ctgggataaa gggcgagatt ttgccacagt gcgcaaagta
     8161 ttgtccatgc cccaagtcaa tattgtcaag aaaacagaag tacagacagg cggattctcc
     8221 aaggagtcaa ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa aaaagactgg
     8281 gatccaaaaa aatatggtgg ttttgatagt ccaacggtag cttattcagt cctagtggtt
     8341 gctaaggtgg aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt actagggatc
     8401 acaattatgg aaagaagttc ctttgaaaaa aatccgattg actttttaga agctaaagga
     8461 tataaggaag ttaaaaaaga cttaatcatt aaactaccta aatatagtct ttttgagtta
     8521 gaaaacggtc gtaaacggat gctggctagt gccggagaat tacaaaaagg aaatgagctg
     8581 gctctgccaa gcaaatatgt gaatttttta tatttagcta gtcattatga aaagttgaag
     8641 ggtagtccag aagataacga acaaaaacaa ttgtttgtgg agcagcataa gcattattta
     8701 gatgagatta ttgagcaaat cagtgaattt tctaagcgtg ttattttagc agatgccaat
     8761 ttagataaag ttcttagtgc atataacaaa catagagaca aaccaatacg tgaacaagca
     8821 gaaaatatta ttcatttatt tacgttgacg aatcttggag ctcccgctgc ttttaaatat
     8881 tttgatacaa caattgatcg taaacgatat acgtctacaa aagaagtttt agatgccact
     8941 cttatccatc aatccatcac tggtctttat gaaacacgca ttgatttgag tcagctagga
     9001 ggtgacggca gcggtgatta taaagatgat gatgataaat gagatatcat tcaggacgag
     9061 cctcagactc cagcgtaact ggactggcgg aacccctatt tgtttatttt tctaaataca
     9121 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     9181 aaggaagagt atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt
     9241 tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc
     9301 agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag
     9361 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc
     9421 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat
     9481 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat
     9541 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa
     9601 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt
     9661 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc
     9721 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc
     9781 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc
     9841 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga
     9901 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat
     9961 caccaaggta gtcggcaaat aactgtcaga ccaagtttac tcatatatac tttagattga
    10021 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
    10081 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg ttgatgatac
    10141 cgctgcctta ctgggtgcat tagccagtct gaatgacctg tcacgggata atccgaagtg
    10201 gtcagactgg aaaatcagag ggcaggaact gctgaacagc aaaaagtcag atagcaccac
    10261 atagcagacc cgccataaaa cgccctgaga agcccgtgac gggcttttct tgtattatgg
    10321 gtagtttcct tgcatgaatc cataaaaggc gcctgtagtg ccatttaccc ccattcactg
    10381 ccagagccgt gagcgcagcg aactgaatgt cacgaaaaag acagcgactc aggtgcctga
    10441 tggtcggaga caaaaggaat attcagcgat ttgcccgatt gcggccgcaa ccgagcttgc
    10501 gagggtgcta cttaagcctt tagggtttta aggtctgttt tgtagaggag caaacagcgt
    10561 ttgcgacatc cttttgtaat actgcggaac tgactaaagt agtgagttat acacagggct
    10621 gggatctatt ctttttatct ttttttattc tttctttatt ctataaatta taaccacttg
    10681 aatataaaca aaaaaaacac acaaaggtct agcggaattt acagagggtc tagcagaatt
    10741 tacaagtttt ccagcaaagg tctagcagaa tttacagata cccacaactc aaaggaaaag
    10801 gactagtaat tatcattgac tagcccatct caattggtat agtgattaaa atcacctaga
    10861 ccaattgaga tgtatgtctg aattagttgt tttcaaagca aatgaactag cgattagtcg
    10921 ctatgactta acggagcatg aaaccaagct aattttatgc tgtgtggcac tactcaaccc
    10981 cacgattgaa aaccctacaa ggaaagaacg gacggtatcg ttcacttata accaatacgt
    11041 tcagatgatg aacatcagta gggaaaatgc ttatggtgta ttagctaaag caaccagaga
    11101 gctgatgacg agaactgtgg aaatcaggaa tcctttggtt aaaggctttg agattttcca
    11161 gtggacaaac tatgccaagt tctcaagcga aaaattagaa ttagttttta gtgaagagat
    11221 attgccttat cttttccagt taaaaaaatt cataaaatat aatctggaac atgttaagtc
    11281 ttttgaaaac aaatactcta tgaggattta tgagtggtta ttaaaagaac taacacaaaa
    11341 gaaaactcac aaggcaaata tagagattag ccttgatgaa tttaagttca tgttaatgct
    11401 tgaaaataac taccatgagt ttaaaaggct taaccaatgg gttttgaaac caataagtaa
    11461 agatttaaac acttacagca atatgaaatt ggtggttgat aagcgaggcc gcccgactga
    11521 tacgttgatt ttccaagttg aactagatag acaaatggat ctcgtaaccg aacttgagaa
    11581 caaccagata aaaatgaatg gtgacaaaat accaacaacc attacatcag attcctacct
    11641 acataacgga ctaagaaaaa cactacacga tgctttaact gcaaaaattc agctcaccag
    11701 ttttgaggca aaatttttga gtgacatgca aagtaagtat gatctcaatg gttcgttctc
    11761 atggctcacg caaaaacaac gaaccacact agagaacata ctggctaaat acggaaggat
    11821 ctgag
//
