LOCUS       sequence-218700-       13694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hSpCas9 under nanos promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13694)
  AUTHORS   Kandul NP, Liu J, Sanchez C HM, Wu SL, Marshall JM, Akbari OS
  TITLE     Transforming insect population control with precision guided sterile
            males with demonstration in flies.
  JOURNAL   Nat Commun. 2019 Jan 8;10(1):84. doi: 10.1038/s41467-018-07964-7.
  PUBMED    30622266
REFERENCE   2  (bases 1 to 13694)
  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: "2019-01-8"; volume: "10(1):84. doi"; pages: " 
            10.1038/s41467-018-07964-7"
COMMENT     Sequence Label: nos-Cas9.874Z1
FEATURES             Location/Qualifiers
     source          1..13694
                     /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   3013..3075
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4109..4178
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     CDS             5315..5380
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5387..5407
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5432..9532
                     /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             5432..9532
                     /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             9533..9580
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9581..9634
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(9635..9637,9638..9640,9641..10354)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTSKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9941..9960)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10288..10309
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(11589..11591,11592..11594,11595..12269)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(11772..11792)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     11990..12009
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     12184..12207
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    12383..12504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12420..12439)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12474..12493
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   13172..13206
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(13387..13403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13387..13403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13400..13422)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13411..13427
                     /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(13435..13441,13442..13459,13460..13465))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13480..13501
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13618..13635)
                     /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 gctttctcat agctcacgct
      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 agaagaacag
      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 tttaaaaata taacaaaatt
     2341 gtgatcccac aaaatgaagt ggggcaaaat caaataatta atagtgtccg taaacttgtt
     2401 ggtcttcaac tttttgagga acacgttgga cggcaaatcc gtgactataa cacaagttga
     2461 tttaataatt ttagccaaca cgtcgggctg cgtgtttttt gccgacgcgt ctgtgtacac
     2521 gttgattaac tggtcgatta aactgttgaa ataatttaat ttttggttct tctttaaatc
     2581 tgtgatgaaa ttttttaaaa taactttaaa ttcttcattg gtaaaaaatg ccacgttttg
     2641 caacttgtga gggtctaata tgaggtcaaa ctcagtagga gttttatcca aaaaagaaaa
     2701 catgattacg tctgtacacg aacgcgtatt aacgcagagt gcaaagtata agagggttaa
     2761 aaaatatatt ttacgcacca tatacgcatc gggttgatat cgttaatatg gatcaatttg
     2821 aacagttgat taacgtgtct ctgctcaagt ctttgatcaa aacgcaaatc gacgaaaatg
     2881 tgtcggacaa tatcaagtcg atgagcgaaa aactaaaaag gctagaatac gacaatctca
     2941 cagacagcgt tgagatatac ggtattcacg acagcaggct gaataataaa aaaattagaa
     3001 actattattt aaccctagaa agataatcat attgtgacgt acgttaaaga taatcatgcg
     3061 taaaattgac gcatgtgttt tatcggtctg tatatcgagg tttatttatt aatttgaata
     3121 gatattaagt tttattatat ttacacttac atactaataa taaattcaac aaacaattta
     3181 tttatgttta tttatttatt aaaaaaaaac aaaaactcaa aatttcttct ataaagtaac
     3241 aaaactttta aacattctct cttttacaaa aataaactta ttttgtactt taaaaacagt
     3301 catgttgtat tataaaataa gtaattagct taacttatac ataatagaaa caaattatac
     3361 ttattagtca gtcagaaaca actttggcac atatcaatat tatgctctcg acaaataact
     3421 tttttgcatt ttttgcacga tgcatttgcc tttcgcctta ttttagaggg gcagtaagta
     3481 cagtaagtac gttttttcat tactggctct tcagtactgt catctgatgt accaggcact
     3541 tcatttggca aaatattaga gatattatcg cgcaaatatc tcttcaaagt aggagcttct
     3601 aaacgcttac gcataaacga tgacgtcagg ctcatgtaaa ggtttctcat aaattttttg
     3661 cgactttgaa ccttttctcc cttgctactg acattatggc tgtatataat aaaagaattt
     3721 atgcaggcaa tgtttatcat tccgtacaat aatgccatag gccacctatt cgtcttccta
     3781 ctgcaggtca tcacagaaca catttggtct agcgtgtcca ctccgccttt agtttgatta
     3841 taatacataa ccatttgcgg tttaccggta ctttcgttga tagaagcatc ctcatcacaa
     3901 gatgataata agtataccat cttagctggc ttcggtttat atgagacgag agtaaggggt
     3961 ccgtcaaaac aaaacatcga tgttcccact ggcctggagc gactgttttt cagtacttcc
     4021 ggtatctcgc gtttgtttga tcgcacggtt cccacaatgg ttaattcgag ctcgcccggg
     4081 gtcctaggtc gacgatgtag gtcacggtct cgaagccgcg gtgcgggtgc cagggcgtgc
     4141 ccttgggctc cccgggcgcg tactccacct cacccatctg gtccatcatg atgaacgggt
     4201 cgaggtggcg gtagttgatc ccggcgaacg cgcggcgcac cgggaagccc tcgccctcga
     4261 aaccgctggg cgcggtggtc acggtgagca cgggacgtgc gacggcgtcg gcgggtgcgg
     4321 atacgcgggg cagcgtcagc gggttctcga cggtcacggc gggcatgtcg acgcaaagct
     4381 tcgaccgttt taacctcgaa atatgcacat gtaaggacgg atgtgagcga acgccagtga
     4441 tgaccgggat cagaggtaac ctaccatggt ggggattagg tgaccgttcg caggtagttt
     4501 gatcggagcg aatgttcggg gggtctggcg tcagaggctc taaactttat gtaattcctg
     4561 ccgcgaaaca cgcacgtatc aagcagtcag ctgttctctt cgttcagcgc gcgccggtgt
     4621 tgcaaaacga gcgctcttcg ccggcggtgg ctcgtgcgat agttcgtttt gtcggtaatc
     4681 cgatgttgcc gcgccgatat catgtgatgt tgtcacagtg cgcgaaattc gaatggtggt
     4741 gtgcagtgat tgtgttgtga cggcgagtgg cgcgtgtggg tgcttagttt tgggagatgt
     4801 tttcgtattt ttttgttgat aactcaggct ttgttgctgt gttgtagtac tattttccat
     4861 tgcgcggtgt ccagctttta attagtggca catattctta gcaagtaaaa attattttgc
     4921 atactattaa atttcttata aattattttc taaaattaag tttacctttt caattttact
     4981 aaaaatatcg atatatttat tatcgctgga aaactacatt attccacctc taagcaagaa
     5041 ccgttagttg gcgcgtagct ttaccacaaa attcctggaa ttgccgtacg cttcgcagtt
     5101 gtttcaagtt gtctaaggga catacgattt tttttgcctc tgcgtcacga ttttaaccca
     5161 aaagcgagtt tagttacatg tacattatta ttagataaag aagtatcgcg aatacttcag
     5221 ttgaataaac tgtgcttggt ttttgggtga ggatttgtgg aaagtagagt gcgcgataac
     5281 cgtaactttc gacccggatt ttcgcctcga gatggactat aaggaccacg acggagacta
     5341 caaggatcat gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg
     5401 gaaggtcggt atccacggag tcccagcagc cgacaagaag tacagcatcg gcctggacat
     5461 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
     5521 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
     5581 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
     5641 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
     5701 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
     5761 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
     5821 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
     5881 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
     5941 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
     6001 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat
     6061 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg
     6121 cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa
     6181 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta
     6241 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct
     6301 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga
     6361 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga
     6421 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt
     6481 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga
     6541 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt
     6601 gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc
     6661 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc
     6721 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta
     6781 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga
     6841 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt
     6901 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca
     6961 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac
     7021 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct
     7081 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa
     7141 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct
     7201 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga
     7261 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat
     7321 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct
     7381 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg
     7441 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag
     7501 aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc
     7561 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc
     7621 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat
     7681 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca
     7741 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct
     7801 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct
     7861 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa
     7921 ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc
     7981 catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc
     8041 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct
     8101 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact
     8161 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt
     8221 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg
     8281 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca
     8341 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc
     8401 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg
     8461 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa
     8521 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac
     8581 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga
     8641 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca
     8701 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct
     8761 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta
     8821 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa
     8881 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag
     8941 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa
     9001 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa
     9061 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa
     9121 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga
     9181 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga
     9241 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct
     9301 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca
     9361 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat
     9421 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag
     9481 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc
     9541 ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag gagggcagag gaagtcttct
     9601 aacatgcggt gacgtggagg agaatcccgg ccctatggtg agcaagggcg aggagctgtt
     9661 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     9721 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     9781 caccaccagc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     9841 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     9901 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     9961 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
    10021 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
    10081 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
    10141 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
    10201 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
    10261 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
    10321 gatcactctc ggcatggacg agctgtacaa gtaattaatt aactagaatg aatcgttttt
    10381 aaaataacaa atcaattgtt ttataatatt cgtacgattc tttgattatg taataaaatg
    10441 tgatcattag gaagattacg aaaaatataa aaaatatgag ttctgtgtgt ataacaaatg
    10501 ctgtaaacgc cacaattgtg tttgttgcaa ataaacccat gattatttga ttaaaattgt
    10561 tgttttcttt gttcatagac aatagtgtgt tttgcctaaa cgtgtactgc ataaactcca
    10621 tgcgagtgta tagcgagcta gtggctaacg cttgccccac caaagtagat tcgtcaaaat
    10681 cctcaatttc atcaccctcc tccaagttta acatttggcc gtcggaatta acttctaaag
    10741 atgccacata atctaataaa tgaaatagag attcaaacgt ggcgtcatcg tccgtttcga
    10801 ccatttccga aaagaactcg ggcataaact ctatgatttc tctggacgtg gtgttgtcga
    10861 aactctcaaa gtacgcagtc aggaacgtgc gcgacatgtc gtcgggaaac tcgcgcggaa
    10921 acatgttgtt gtaaccgaac gggtcccata gcgccaaaac caaatctgcc agcgtcaata
    10981 gaatgagcac gatgccgaca atggagctgg cttggatagc gattcgagtt aacggccggg
    11041 gcgcgcccat actcggtggc ctccccacca ccaacttttt tgcactgcaa aaaaacacgc
    11101 ttttgcacgc gggcccatac atagtacaaa ctctacgttt cgtagactat tttacataaa
    11161 tagtctacac cgttgtatac gctccaaata cactaccaca cattgaacct ttttgcagtg
    11221 caaaaaagta cgtgtcggca gtcacgtagg ccggccttat cgggtcgcgt cctgtcacgt
    11281 acgaatcaca ttatcggacc ggacgagtgt tgtcttatcg tgacaggacg ccagcttcct
    11341 gtgttgctaa ccgcagccgg acgcaactcc ttatcggaac aggacgcgcc tccatatcag
    11401 ccgcgcgtta tctcatgcgc gtgaccggac acgaggcgcc cgtcccgctt atcgcgccta
    11461 taaatacagc ccgcaacgat ctggtaaaca cagttgaaca gcatctgtta cagcgacaca
    11521 acatgagccg gtccaacaac gccaacgcgc ccacgccatc caaccgccgc cgcaacctgt
    11581 ctctggtgat ggtgcgctcc tccaagaacg tcatcaagga gttcatgcgc ttcaaggtgc
    11641 gcatggaggg caccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
    11701 cctacgaggg ccacaacacc gtgaagctga aggtgaccaa gggcggcccc ctgcccttcg
    11761 cctgggacat cctgtccccc cagttccagt acggctccaa ggtgtacgtg aagcaccccg
    11821 ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
    11881 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggct
    11941 gcttcatcta caaggtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc
    12001 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
    12061 tgaagggcga gatccacaag gccctgaagc tgaaggacgg cggccactac ctggtggagt
    12121 tcaagtccat ctacatggcc aagaagcccg tgcagctgcc cggctactac tacgtggact
    12181 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggagcag tacgagcgca
    12241 ccgagggccg ccaccacctg ttcctgtagg actctagatc ataatcagcc ataccacatt
    12301 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
    12361 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
    12421 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    12481 gtccaaactc atcaatgtat cttaaagctt ctaggccggc cgatctcgga tctgacaatg
    12541 ttcagtgcag agactcggct acgcctcgtg gactttgaag ttgaccaaca atgtttattc
    12601 ttacctctaa tagtcctctg tggcaaggtc aagattctgt tagaagccaa tgaagaacct
    12661 ggttgttcaa taacattttg ttcgtctaat atttcactac cgcttgacgt tggctgcact
    12721 tcatgtacct catctataaa cgcttcttct gtatcgctct ggacgtcatc ttcacttacg
    12781 tgatctgata tttcactgtc agaatcctca ccaacaagct cgtcatcgct ttgcagaaga
    12841 gcagagagga tatgctcatc gtctaaagaa ctacccattt tattatatat tagtcacgat
    12901 atctataaca agaaaatata tatataataa gttatcacgt aagtagaaca tgaaataaca
    12961 atataattat cgtatgagtt aaatcttaaa agtcacgtaa aagataatca tgcgtcattt
    13021 tgactcacgc ggtcgttata gttcaaaatc agtgacactt accgcattga caagcacgcc
    13081 tcacgggagc tccaagcggc gactgagatg tcctaaatgc acagcgacgg attcgcgcta
    13141 tttagaaaga gagagcaata tttcaagaat gcatgcgtca attttacgca gactatcttt
    13201 ctagggttaa aaaagatttg cgctttactc gacctaaact ttaaacacgt catagaatct
    13261 tcgtttgaca aaaaccacat tgtggccaag ctgtgtgacg cgacgcgcgc taaagaatgg
    13321 caaaccaagt cgcgcgagcg tcgactctag aggatccccg ggtaccgagc tcgaattcgt
    13381 aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
    13441 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
    13501 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
    13561 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
    13621 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
    13681 ggcggtaata cggt
//
