LOCUS       sequence-89645-e       11652 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses human codon-optimized S. pyogenes Cas9 from the human 
            PGK-1 gene promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11652)
  AUTHORS   Maggio I, Holkers M, Liu J, Janssen JM, Chen X, Goncalves MA
  TITLE     Adenoviral vector delivery of RNA-guided CRISPR/Cas9 nuclease 
            complexes induces targeted mutagenesis in a diverse array of human 
            cells.
  JOURNAL   Sci Rep. 2014 May 29;4:5105. doi: 10.1038/srep05105.
  PUBMED    24870050
REFERENCE   2  (bases 1 to 11652)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2014-05-29"; volume: "4:5105. doi"; pages: " 
            10.1038/srep05105"
COMMENT     Sequence Label: pAdSh.PGK.Cas9
FEATURES             Location/Qualifiers
     source          1..11652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..103
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     191..341
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     promoter        454..963
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     883..901
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      994..1003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1000..5103
                     /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             1000..5103
                     /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             1000..5103
                     /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             5116..5136
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5207..5328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5218..5237)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5272..5291
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   8612..8714
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(8803..8820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8974..9562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9054..9073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             10394..11188
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             10394..11188
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(10448..10467)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     11058..11077
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(11236..11615)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11298..11319)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11510..11529
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt
       61 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt
      121 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg
      181 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag
      241 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga
      301 agtgaaatct gaataatttt gtgttactca tagcgcgtaa tactggtacc ggttgttaac
      361 gttagccggc tacgtatact ccggaatatt aataggccta ggatgcatat ggcggccgcc
      421 tgcagctggc gccatcgata cgcgtacttc cacggggttg gggttgcgcc ttttccaagg
      481 cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg
      541 cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc
      601 cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc
      661 cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct
      721 cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa
      781 tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc
      841 ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa
      901 gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc
      961 ccaccgggta cgtcgctaga ggatcgaacc cttgccacca tggacaagaa gtactccatt
     1021 gggctcgata tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg
     1081 ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc
     1141 attggcgccc tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca
     1201 gcacggcgca gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt
     1261 aatgagatgg ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg
     1321 gaggaggata aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg
     1381 taccatgaaa agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat
     1441 aaggctgact tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac
     1501 ttcctcatcg agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa
     1561 ctggttcaga cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac
     1621 gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca
     1681 cagctccctg gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg
     1741 ctgaccccca actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc
     1801 aaagacacct acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca
     1861 gacctttttt tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga
     1921 gtgaacacgg agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag
     1981 caccaccaag acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac
     2041 aaggaaattt tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca
     2101 agccaggagg aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag
     2161 gagctgctgg taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat
     2221 ggaagcatcc cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag
     2281 gatttctacc cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg
     2341 ataccctact atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc
     2401 aaatcagaag agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct
     2461 gcccagtcct tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg
     2521 cttcctaaac actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc
     2581 aaatacgtca cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct
     2641 atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac
     2701 tatttcaaaa agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc
     2761 aacgcatccc tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg
     2821 gacaatgagg agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa
     2881 gatagggaga tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc
     2941 atgaaacagc tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc
     3001 aatgggatcc gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga
     3061 tttgccaacc ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac
     3121 atccagaaag cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt
     3181 gcaggtagcc cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc
     3241 gtcaaagtaa tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac
     3301 caaactaccc agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt
     3361 ataaaagaac tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag
     3421 aatgagaagc tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa
     3481 ctggacatca atcggctctc cgactacgac gtggatcata tcgtgcccca gtcttttctc
     3541 aaagatgatt ctattgataa taaagtgttg acaagatccg ataaaaatag agggaagagt
     3601 gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg
     3661 aacgccaaac tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc
     3721 ctgtctgagt tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc
     3781 accaagcacg tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac
     3841 aaactgattc gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga
     3901 aaggactttc agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc
     3961 tacctgaatg cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa
     4021 tttgtttacg gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag
     4081 gaaataggca aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag
     4141 accgagatta cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga
     4201 gaaacaggag aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg
     4261 tccatgccgc aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag
     4321 gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac
     4381 cccaagaaat acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc
     4441 aaagtggaga aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca
     4501 atcatggagc gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat
     4561 aaagaggtca aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa
     4621 aacggccgga aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca
     4681 ctgccctcta aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg
     4741 tctcccgaag ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat
     4801 gagatcatcg agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc
     4861 gataaggtgc tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa
     4921 aacattatcc acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc
     4981 gacaccacca tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg
     5041 attcatcagt caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga
     5101 gacagcaggg ctgaccccaa gaagaagagg aaggtgtgaa agggttcgat ccctacccga
     5161 ccgcggacat gtacagagct cgagcggccg cttcgagcag acatgataag atacattgat
     5221 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     5281 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     5341 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     5401 aacctctaca aatgtggtaa aatcgataag gatctgggcg tggttaaggg tgggaaagaa
     5461 tatataaggt gggggtctta tgtagttttg tatctgtttt gcagcagccg ccgccgccat
     5521 gagcaccaac tcgtttgatg gaagcattgt gagctcatat ttgacaacgc gcatgccccc
     5581 atgggccggg gtgcgtcaga atgtgatggg ctccagcatt gatggtcgcc ccgtcctgcc
     5641 cgcaaactct actaccttga cctacgagac cgtgtctgga acgccgttgg agactgcagc
     5701 ctccgccgcc gcttcagccg ctgcagccac cgcccgcggg attgtgactg actttgcttt
     5761 cctgagcccg cttgcaagca gtgcagcttc ccgttcatcc gcccgcgatg acaagttgac
     5821 ggctcttttg gcacaattgg attctttgac ccgggaactt aatgtcgttt ctcagcagct
     5881 gttggatctg cgccagcagg tttctgccct gaaggcttcc tcccctccca atgcggttta
     5941 aaacataaat aaaaaaccag actctgtttg gatttggatc aagcaagtgt cttgctgtct
     6001 ttatttaggg gttttgcgcg cgcggtaggc ccgggaccag cggtctcggt cgttgagggt
     6061 cctgtgtatt ttttccagga cgtggtaaag gtgactctgg atgttcagat acatgggcat
     6121 aagcccgtct ctggggtgga ggtagcacca ctgcagagct tcatgctgcg gggtggtgtt
     6181 gtagatgatc cagtcgtagc aggagcgctg ggcgtggtgc ctaaaaatgt ctttcagtag
     6241 caagctgatt gccaggggca ggcccttggt gtaagtgttt acaaagcggt taagctggga
     6301 tgggtgcata cgtggggata tgagatgcat cttggactgt atttttaggt tggctatgtt
     6361 cccagccata tccctccggg gattcatgtt gtgcagaacc accagcacag tgtatccggt
     6421 gcacttggga aatttgtcat gtagcttaga aggaaatgcg tggaagaact tggagacgcc
     6481 cttgtgacct ccaagatttt ccatgcattc gtccataatg atggcaatgg gcccacgggc
     6541 ggcggcctgg gcgaagatat ttctgggatc actaacgtca tagttgtgtt ccaggatgag
     6601 atcgtcatag gccattttta caaagcgcgg gcggagggtg ccagactgcg gtataatggt
     6661 tccatccggc ccaggggcgt agttaccctc acagatttgc atttcccacg ctttgagttc
     6721 agatgggggg atcatgtcta cctgcggggc gatgaagaaa acggtttccg gggtagggga
     6781 gatcagctgg gaagaaagca ggttcctgag cagctgcgac ttaccgcagc cggtgggccc
     6841 gtaaatcaca cctattaccg ggtgcaactg gtagttaaga gagctgcagc tgccgtcatc
     6901 cctgagcagg ggggccactt cgttaagcat gtccctgact cgcatgtttt ccctgaccaa
     6961 atccgccaga aggcgctcgc cgcccagcga tagcagttct tgcaaggaag caaagttttt
     7021 caacggtttg agaccgtccg ccgtaggcat gcttttgagc gtttgaccaa gcagttccag
     7081 gcggtcccac agctcggtca cctgctctac ggcatctcga tccagcatat ctcctcgttt
     7141 cgcgggttgg ggcggctttc gctgtacggc agtagtcggt gctcgtccag acgggccagg
     7201 gtcatgtctt tccacgggcg cagggtcctc gtcagcgtag tctgggtcac ggtgaagggg
     7261 tgcgctccgg gctgcgcgct ggccagggtg cgcttgaggc tggtcctgct ggtgctgaag
     7321 cgctgccggt cttcgccctg cgcgtcggcc aggtagcatt tgaccatggt gtcatagtcc
     7381 agcccctccg cggcgtggcc cttggcgcgc agcttgccct tggaggaggc gccgcacgag
     7441 gggcagtgca gacttttgag ggcgtagagc ttgggcgcga gaaataccga ttccggggag
     7501 taggcatccg cgccgcaggc cccgcagacg gtctcgcatt ccacgagcca ggtgagctct
     7561 ggccgttcgg ggtcaaaaac caggtttccc ccatgctttt tgatgcgttt cttacctctg
     7621 gtttccatga gccggtgtcc acgctcggtg acgaaaaggc tgtccgtgtc cccgtataca
     7681 gacttgagag ggagtttaaa cgaattcaat agcttgttgc atgggcggcg atataaaatg
     7741 caaggtgctg ctcaaaaaat caggcaaagc ctcgcgcaaa aaagaaagca catcgtagtc
     7801 atgctcatgc agataaaggc aggtaagctc cggaaccacc acagaaaaag acaccatttt
     7861 tctctcaaac atgtctgcgg gtttctgcat aaacacaaaa taaaataaca aaaaaacatt
     7921 taaacattag aagcctgtct tacaacagga aaaacaaccc ttataagcat aagacggact
     7981 acggccatgc cggcgtgacc gtaaaaaaac tggtcaccgt gattaaaaag caccaccgac
     8041 agctcctcgg tcatgtccgg agtcataatg taagactcgg taaacacatc aggttgattc
     8101 atcggtcagt gctaaaaagc gaccgaaata gcccggggga atacataccc gcaggcgtag
     8161 agacaacatt acagccccca taggaggtat aacaaaatta ataggagaga aaaacacata
     8221 aacacctgaa aaaccctcct gcctaggcaa aatagcaccc tcccgctcca gaacaacata
     8281 cagcgcttca cagcggcagc ctaacagtca gccttaccag taaaaaagaa aacctattaa
     8341 aaaaacacca ctcgacacgg caccagctca atcagtcaca gtgtaaaaaa gggccaagtg
     8401 cagagcgagt atatatagga ctaaaaaatg acgtaacggt taaagtccac aaaaaacacc
     8461 cagaaaaccg cacgcgaacc tacgcccaga aacgaaagcc aaaaaaccca caacttcctc
     8521 aaatcgtcac ttccgttttc ccacgttacg taacttccca ttttaagaaa actacaattc
     8581 ccaacacata caagttactc cgccctaaaa cctacgtcac ccgccccgtt cccacgcccc
     8641 gcgccacgtc acaaactcca ccccctcatt atcatattgg cttcaatcca aaataaggta
     8701 tattattgat gatgttaatt aacatgcatg gatccatatg cggtgtgaaa taccgcacag
     8761 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     8821 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     8881 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     8941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     9001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     9061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     9121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     9181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     9241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     9301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     9361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     9421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     9481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     9541 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     9601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     9661 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     9721 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     9781 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     9841 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     9901 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     9961 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    10021 tagtttgcgc aacgttgttg ccattgctgc agccatgaga ttatcaaaaa ggatcttcac
    10081 ctagatcctt ttcacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt
    10141 cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg
    10201 cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga
    10261 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
    10321 tttcttgccg ccaaggatct gatggcgcag gggatcaagc tctgatcaag agacaggatg
    10381 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
    10441 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
    10501 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
    10561 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
    10621 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
    10681 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
    10741 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
    10801 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
    10861 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
    10921 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
    10981 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
    11041 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
    11101 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
    11161 ctatcgcctt cttgacgagt tcttctgaat tttgttaaaa tttttgttaa atcagctcat
    11221 tttttaacca ataggccgaa atcggcaaca tcccttataa atcaaaagaa tagaccgcga
    11281 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
    11341 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
    11401 aatcaagttt tttgcggtcg aggtgccgta aagctctaaa tcggaaccct aaagggagcc
    11461 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
    11521 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
    11581 cacccgcgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gatcgaatta
    11641 attcttaatt aa
//
