LOCUS       sequence-320195-       15056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AAEL005635-Cas9 with OpIE2-CFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15056)
  AUTHORS   Li M, Yang T, Bui M, Gamez S, Wise T, Kandul NP, Liu J, Alcantara L,
            Lee H, Edula JR, Raban R, Zhan Y, Wang Y, DeBeaubien N, Chen J, 
            Sanchez C HM, Bennett JB, Antoshechkin I, Montell C, Marshall JM, 
            Akbari OS
  TITLE     Suppressing mosquito populations with precision guided sterile 
            males.
  JOURNAL   Nat Commun. 2021 Sep 10;12(1):5374. doi: 10.1038/s41467-021-25421-w.
  PUBMED    34508072
REFERENCE   2  (bases 1 to 15056)
  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: "Nat 
            Commun."; date: "2021-09-10"; volume: "12(1):5374. doi"; pages: " 
            10.1038/s41467-021-25421-w"
COMMENT     Sequence Label: OA-874PA
FEATURES             Location/Qualifiers
     source          1..15056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(95..683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(175..194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(854..1645,1646..1714))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1477..1496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1715..1819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1887..1905
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1943..1965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2065..2084
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2278..2300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2292..2309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2293..2309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2998..3060
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4094..4163
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     CDS             7409..7474
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7481..7501
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7526..11626
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7526..11626
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             11627..11674
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(12921..12923,12924..12926,12927..13640)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(13227..13246)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     13574..13595
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    13753..13874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13790..13809)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13844..13863
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   14533..14567
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(14749..14765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14749..14765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14762..14784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14773..14789
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(14797..14803,14804..14821,14822..14827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14842..14863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14980..14997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
       61 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
      301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
      541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
      841 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
      961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1861 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1921 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1981 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2041 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2101 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2161 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2221 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2281 agtcacgacg ttgtaaaacg acggccagtg ccaagctttg tttaaaatat aacaaaattg
     2341 tgatcccaca aaatgaagtg gggcaaaatc aaataattaa ctagtgtccg taaacttgtt
     2401 ggtcttcaac tttttgagga acacgttgga cggcaaatcg tgactataac acaagttgat
     2461 ttaataattt tagccaacac gtcgggctgc gtgttttttg cgctctgtgt acacgttgat
     2521 taactggtcg attaaataat ttaatttttg gttcttcttt aaatctgtga tgaaattttt
     2581 taaaataact ttaaattctt cattggtaaa aaatgccacg ttttgcaact tgtgagggtc
     2641 taatatgagg tcaaactcag taggagtttt atccaaaaaa gaaaacatga ttacgtctgt
     2701 acacgaacgc gtattaacgc agagtgcaaa gtataagagg gttaaaaaat atattttacg
     2761 caccatatac gcatcgggtt gatatcgtta atatggatca atttgaacag ttgattaacg
     2821 tgtctctgct caagtctttg atcaaaacgc aaatcgacga aaatgtgtcg gacaatatca
     2881 agtcgatgag cgaaaaacta aaaaggctag aatacgacaa tctcacagac agcgttgaga
     2941 tatacggtat tcacgacagc aggctgaata ataaaaaaat tagaaactat tatttaaccc
     3001 tagaaagata atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg
     3061 tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat
     3121 tatatttaca cttacatact aataataaat tcaacaaaca atttatttat gtttatttat
     3181 ttattaaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttaaacat
     3241 tctctctttt acaaaaataa acttattttg tactttaaaa acagtcatgt tgtattataa
     3301 aataagtaat tagcttaact tatacataat agaaacaaat tatacttatt agtcagtcag
     3361 aaacaacttt ggcacatatc aatattatgc tctcgacaaa taactttttt gcattttttg
     3421 cacgatgcat ttgcctttcg ccttatttta gaggggcagt aagtacagta agtacgtttt
     3481 ttcattactg gctcttcagt actgtcatct gatgtaccag gcacttcatt tggcaaaata
     3541 ttagagatat tatcgcgcaa atatctcttc aaagtaggag cttctaaacg cttacgcata
     3601 aacgatgacg tcaggctcat gtaaaggttt ctcataaatt ttttgcgact ttgaaccttt
     3661 tctcccttgc tactgacatt atggctgtat ataataaaag aatttatgca ggcaatgttt
     3721 atcattccgt acaataatgc cataggccac ctattcgtct tcctactgca ggtcatcaca
     3781 gaacacattt ggtctagcgt gtccactccg cctttagttt gattataata cataaccatt
     3841 tgcggtttac cggtactttc gttgatagaa gcatcctcat cacaagatga taataagtat
     3901 accatcttag ctggcttcgg tttatatgag acgagagtaa ggggtccgtc aaaacaaaac
     3961 atcgatgttc ccactggcct ggagcgactg tttttcagta cttccggtat ctcgcgtttg
     4021 tttgatcgca cggttcccac aatggttaat tcgagctcgc ccggggtcct aggtcgacga
     4081 tgtaggtcac ggtctcgaag ccgcggtgcg ggtgccaggg cgtgcccttg ggctccccgg
     4141 gcgcgtactc cacctcaccc atctggtcca tcatgatgaa cgggtcgagg tggcggtagt
     4201 tgatcccggc gaacgcgcgg cgcaccggga agccctcgcc ctcgaaaccg ctgggcgcgg
     4261 tggtcacggt gagcacggga cgtgcgacgg cgtcggcggg tgcggatacg cggggcagcg
     4321 tcagcgggtt ctcgacggtc acggcgggca tgtcgacgcg gccgcatgtg aatcaacatt
     4381 aggttgtcat gagaaacaaa catgaatgtc taaatattaa atcatgaaaa tgaaaaccaa
     4441 ctgatttgct tattaaattc taaatgtagt ggctttagat agtcatgtgt gatagaacat
     4501 gaatgtcatg agactgaaat aagaaatttg atagaagaaa tcttgctccc caaaatatgg
     4561 attcgaggct cctctgcttg tcgcaagctc acttgaattt ttttttttaa aacaaatgag
     4621 tcagcaacaa gcggagcgcc gcaaatccat aatttgtgaa gtctgggatt agcatgtttc
     4681 attttattcg aagctgaaag ctaggaaaat ctgttagtgg aatccgattt ttctctaaca
     4741 ccatgcatta catccaacgc tggaagtaat gcattttatt tatagaaaat tagaatgaac
     4801 tcctcatatt cactcggaca tcttcactaa tagaaagaaa aatgactccg caaattttcc
     4861 tctaacactt tcagcttcag aatttgcctc tttactttgg aacatgatga tgactgtcgt
     4921 tcgacatgag gtccatccgc aatttagttc gctatgggtt gatcactacc caagtaacca
     4981 ataagctgtt tatatcggca ccaagtcggt tctatagtcg atttaaaggc ttggtcgaca
     5041 gagccaattg gaactatatc gtaataaacg gctgttataa agccactttt ggcgaattat
     5101 ttggctgata aacagcctac tcgtactgtg tctctgaaat gagtgatgtc aaaattttat
     5161 agcaagtggc aaaaaaaaga gaaaaaaaag ttgtgtatct tctgttcgat tttcatttct
     5221 tttatgctcc ctctggagac gttttttaat ttcaggctca aactgctgtc gtctaagttc
     5281 atattaatga ttcccgtcgc attttctatc tgctccatcg atgcacgagt ggattgggat
     5341 gcttaacgtc gtatttaaaa gagaaatgtt tgcagcgatg tgtctatttt tgttctgctg
     5401 aatcacgagt gatgctggcg atgccggcaa ggcatttgac catatatgta tagcagcaca
     5461 aaagaagcga aatcaacaga aaaaaaatta caatcaattt acgaatctgt tcacatcttt
     5521 gagcatctta aattaaaaca ttcctgataa cgccgtttgt tttgttttct gcggtgagag
     5581 aaaaagaggc ggagagatgt gtttcgctcc attccctcgc atgttgacat ttgcgtacga
     5641 tatgaaatct ctttcgccgt aatcccgatt cagttttgtt ttggttgata aacgtcatga
     5701 cactaggccg tttagaagca ttgacggtcg ttgtgtacgg cttattggca cagggcttac
     5761 aagcactaaa ataaagcgtt atatactcat atacggccaa tatcagtgcc tgatggtcac
     5821 agtgcctata agcattcgaa aatgttttat gaagctagta ggcactgaaa atccgttaaa
     5881 tgtctcatat accgcttata acagtgcttg ttggttactt cttcttcttc tttctggcgt
     5941 tacgtcccca ctgggacaga gcctgcttct cggcttagtg ttcttatgag cacttccaca
     6001 gtaattatct gagagcttac tatgccaatg accatttttg catgtgtata tcgtgtggca
     6061 ggtacgaaga tactctatgc cctgggaaat cgagaaaatt tccaacccga aaatatcctc
     6121 gaccggtggg attcgaaccc acgaccctca gcttggtctt gctgaatagc tgcgcgttta
     6181 ccgctacggc tatctgggcc cctgggtaat gattcagaaa tttggaacac ggcaatcgac
     6241 agcaagctca tcgatttcaa atcgaccaac ttgaatatga tgatcatgac acaagataat
     6301 cataagttaa acacactgaa tccgtgttgg gtgatgatcc atagcaccca taggttgacg
     6361 catacgaatt tgaaatggaa agctttagga aatcatgtgg aaaactatgg aattcatgtt
     6421 gtcggttgat gaatcagtcc atgtagcaaa actaaagaat tgaatgtgta atacacatga
     6481 agtttgtaag aaaatcacaa caaatcgaca tagacgttca tgaatcgatc cataatttta
     6541 aacacacgga tcaagcgtgt ggattttttt cagtgggggg tgggtgggtg tcctcgtggt
     6601 gttacggctc atacaaaatt tgtaaactat tcatataaaa agcgttacga gggggtgggt
     6661 gggtgtcgaa aatggccatt tttggcgtta tgaaatttgt gaataaaccc atagtttatt
     6721 tcgaatacaa tacttttcac tttcacagcc ataaaaagga cgggctgcat ttgaagtttc
     6781 atgacagtcg aatctggact gcatgggctc attcgcaaat tttataacgc tggagggggt
     6841 ggatgggtgt cctagggttg ttacagctca tacaaaattc gaaaaacatt catacaaaat
     6901 gggttacgag ggggtgggta ggtatggaaa atgaccattt tcggcgttat gaaatttgtt
     6961 gatataccca gacgttgata tatatttttc ctctgcgcga gtctctctca ggttccaacc
     7021 agcgaatccc cgctgtaaca agcttcgact gtactttggc caccagccgc gtaaaagctt
     7081 catgaacgag ctccttcgct ctgatgcatg cgcagtcaca ggggctccag ctgtcaaatc
     7141 atccggcaaa accgaactca aaacaaaacg tgtttgtgtg tttcgtgtgt ttccgcgcgt
     7201 tgctttctct tggataattt cacaaagttt tcgtgaattg tgccgtaaaa atcgtgttaa
     7261 ttcaacagaa tattttctgt aattgtttgc cagacgcaca ctgtttgcac gtaagtaaaa
     7321 atccacaaag attgtcaaaa ttctatcaag tttttaccgc aaagtcttac gtccctcaga
     7381 aggctccacc ctggggaaac tcgagatgga ctataaggac cacgacggag actacaagga
     7441 tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt
     7501 cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac
     7561 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
     7621 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     7681 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     7741 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
     7801 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     7861 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     7921 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     7981 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     8041 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     8101 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     8161 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     8221 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     8281 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     8341 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     8401 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     8461 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     8521 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     8581 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     8641 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     8701 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     8761 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     8821 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     8881 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     8941 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     9001 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     9061 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     9121 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     9181 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     9241 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     9301 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     9361 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     9421 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     9481 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     9541 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     9601 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     9661 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     9721 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     9781 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     9841 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     9901 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     9961 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
    10021 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
    10081 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
    10141 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
    10201 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
    10261 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
    10321 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
    10381 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
    10441 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
    10501 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
    10561 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
    10621 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
    10681 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
    10741 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
    10801 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
    10861 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
    10921 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
    10981 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
    11041 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
    11101 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
    11161 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
    11221 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
    11281 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
    11341 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
    11401 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
    11461 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
    11521 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
    11581 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc
    11641 cacgaaaaag gccggccagg caaaaaagaa aaagtaatta attaactaga atgaatcgtt
    11701 tttaaaataa caaatcaatt gttttataat attcgtacga ttctttgatt atgtaataaa
    11761 atgtgatcat taggaagatt acgaaaaata taaaaaatat gagttctgtg tgtataacaa
    11821 atgctgtaaa cgccacaatt gtgtttgttg caaataaacc catgattatt tgattaaaat
    11881 tgttgttttc tttgttcata gacaatagtg tgttttgcct aaacgtgtac tgcataaact
    11941 ccatgcgagt gtatagcgag ctagtggcta acgcttgccc caccaaagta gattcgtcaa
    12001 aatcctcaat ttcatcaccc tcctccaagt ttaacatttg gccgtcggaa ttaacttcta
    12061 aagatgccac ataatctaat aaatgaaata gagattcaaa cgtggcgtca tcgtccgttt
    12121 cgaccatttc cgaaaagaac tcgggcataa actctatgat ttctctggac gtggtgttgt
    12181 cgaaactctc aaagtacgca gtcaggaacg tgcgcgacat gtcgtcggga aactcgcgcg
    12241 gaaacatgtt gttgtaaccg aacgggtccc atagcgccaa aaccaaatct gccagcgtca
    12301 atagaatgag cacgatgccg acaatggagc tggcttggat agcgattcga gttaacggcc
    12361 ggggcgcgcc catactcggt ggcctcccca ccaccaactt ttttgcactg caaaaaaaca
    12421 cgcttttgca cgcgggccca tacatagtac aaactctacg tttcgtagac tattttacat
    12481 aaatagtcta caccgttgta tacgctccaa atacactacc acacattgaa cctttttgca
    12541 gtgcaaaaaa gtacgtgtcg gcagtcacgt aggccggcct tatcgggtcg cgtcctgtca
    12601 cgtacgaatc acattatcgg accggacgag tgttgtctta tcgtgacagg acgccagctt
    12661 cctgtgttgc taaccgcagc cggacgcaac tccttatcgg aacaggacgc gcctccatat
    12721 cagccgcgcg ttatctcatg cgcgtgaccg gacacgaggc gcccgtcccg cttatcgcgc
    12781 ctataaatac agcccgcaac gatctggtaa acacagttga acagcatctg ttacagcgac
    12841 acaacatgag ccggtccaac aacgccaacg cgcccacgcc atccaaccgc cgccgcaacc
    12901 tgtctctggt gctagccaaa atggtgagca agggcgagga gctgttcacc ggggtggtgc
    12961 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
    13021 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
    13081 tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcagcc
    13141 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
    13201 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
    13261 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
    13321 acggcaacat cctggggcac aagctggagt acaactacat cagccacaac gtctatatca
    13381 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
    13441 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
    13501 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
    13561 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
    13621 tggacgagct gtacaagtaa ggcctagatc ataatcagcc ataccacatt tgtagaggtt
    13681 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
    13741 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    13801 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    13861 atcaatgtat cttaaagctt aagatctcgg atctgacaat gttcagtgca gagactcggc
    13921 tacgcctcgt ggactttgaa gttgaccaac aatgtttatt cttacctcta atagtcctct
    13981 gtggcaaggt caagattctg ttagaagcca atgaagaacc tggttgttca ataacatttt
    14041 gttcgtctaa tatttcacta ccgcttgacg ttggctgcac ttcatgtacc tcatctataa
    14101 acgcttcttc tgtatcgctc tggacgtcat cttcacttac gtgatctgat atttcactgt
    14161 cagaatcctc accaacaagc tcgtcatcgc tttgcagaag agcagagagg atatgctcat
    14221 cgtctaaaga actacccatt ttattatata ttagtcacga tatctataac aagaaaatat
    14281 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
    14341 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
    14401 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
    14461 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
    14521 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta aaaaagattt
    14581 gcgctttact cgacctaaac tttaaacacg tcatagaatc ttcgtttgac aaaaaccaca
    14641 ttgtggccaa gctgtgtgac gcgacgcgcg ctaaagaatg gcaaaccaag tcgcgcgagc
    14701 gtcgactcta gaggatcccc gggtaccgag ctcgaattcg taatcatggt catagctgtt
    14761 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
    14821 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
    14881 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
    14941 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
    15001 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggt
//
