LOCUS       sequence-327852-       15070 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 15070)
  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 15070)
  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..15070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..66
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             73..93
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             118..4218
                     /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             118..4218
                     /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             4219..4266
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(5513..5515,5516..5518,5519..6232)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5819..5838)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6166..6187
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6345..6466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6382..6401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6436..6455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7125..7159
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7340..7356)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7340..7356)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7353..7375)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7364..7380
                     /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(7388..7394,7395..7412,7413..7418))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7433..7454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7571..7588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7742..8330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7822..7841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8501..9292,9293..9361))
                     /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     9124..9143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9362..9466)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9534..9552
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9590..9612)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     9712..9731
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9925..9947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9939..9956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9940..9956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   10660..10722
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    11756..11825
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 gactataagg accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac
       61 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac
      121 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
      181 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
      241 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
      301 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
      361 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
      421 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
      481 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
      541 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
      601 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
      661 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
      721 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
      781 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
      841 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
      901 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
      961 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     1021 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     1081 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     1141 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     1201 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     1261 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     1321 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     1381 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     1441 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     1501 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     1561 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     1621 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     1681 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     1741 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     1801 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     1861 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     1921 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     1981 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     2041 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     2101 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     2161 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     2221 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     2281 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     2341 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     2401 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     2461 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     2521 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     2581 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     2641 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     2701 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     2761 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     2821 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     2881 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     2941 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     3001 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     3061 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     3121 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     3181 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     3241 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     3301 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     3361 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     3421 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     3481 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     3541 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     3601 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     3661 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     3721 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     3781 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     3841 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     3901 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     3961 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     4021 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     4081 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     4141 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     4201 tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     4261 aaaaagtaat taattaacta gaatgaatcg tttttaaaat aacaaatcaa ttgttttata
     4321 atattcgtac gattctttga ttatgtaata aaatgtgatc attaggaaga ttacgaaaaa
     4381 tataaaaaat atgagttctg tgtgtataac aaatgctgta aacgccacaa ttgtgtttgt
     4441 tgcaaataaa cccatgatta tttgattaaa attgttgttt tctttgttca tagacaatag
     4501 tgtgttttgc ctaaacgtgt actgcataaa ctccatgcga gtgtatagcg agctagtggc
     4561 taacgcttgc cccaccaaag tagattcgtc aaaatcctca atttcatcac cctcctccaa
     4621 gtttaacatt tggccgtcgg aattaacttc taaagatgcc acataatcta ataaatgaaa
     4681 tagagattca aacgtggcgt catcgtccgt ttcgaccatt tccgaaaaga actcgggcat
     4741 aaactctatg atttctctgg acgtggtgtt gtcgaaactc tcaaagtacg cagtcaggaa
     4801 cgtgcgcgac atgtcgtcgg gaaactcgcg cggaaacatg ttgttgtaac cgaacgggtc
     4861 ccatagcgcc aaaaccaaat ctgccagcgt caatagaatg agcacgatgc cgacaatgga
     4921 gctggcttgg atagcgattc gagttaacgg ccggggcgcg cccatactcg gtggcctccc
     4981 caccaccaac ttttttgcac tgcaaaaaaa cacgcttttg cacgcgggcc catacatagt
     5041 acaaactcta cgtttcgtag actattttac ataaatagtc tacaccgttg tatacgctcc
     5101 aaatacacta ccacacattg aacctttttg cagtgcaaaa aagtacgtgt cggcagtcac
     5161 gtaggccggc cttatcgggt cgcgtcctgt cacgtacgaa tcacattatc ggaccggacg
     5221 agtgttgtct tatcgtgaca ggacgccagc ttcctgtgtt gctaaccgca gccggacgca
     5281 actccttatc ggaacaggac gcgcctccat atcagccgcg cgttatctca tgcgcgtgac
     5341 cggacacgag gcgcccgtcc cgcttatcgc gcctataaat acagcccgca acgatctggt
     5401 aaacacagtt gaacagcatc tgttacagcg acacaacatg agccggtcca acaacgccaa
     5461 cgcgcccacg ccatccaacc gccgccgcaa cctgtctctg gtgctagcca aaatggtgag
     5521 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     5581 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     5641 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     5701 caccctgacc tggggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     5761 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     5821 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5881 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5941 gtacaactac atcagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     6001 ggccaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     6061 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     6121 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     6181 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaggcctaga
     6241 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     6301 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     6361 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6421 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaagc ttaagatctc
     6481 ggatctgaca atgttcagtg cagagactcg gctacgcctc gtggactttg aagttgacca
     6541 acaatgttta ttcttacctc taatagtcct ctgtggcaag gtcaagattc tgttagaagc
     6601 caatgaagaa cctggttgtt caataacatt ttgttcgtct aatatttcac taccgcttga
     6661 cgttggctgc acttcatgta cctcatctat aaacgcttct tctgtatcgc tctggacgtc
     6721 atcttcactt acgtgatctg atatttcact gtcagaatcc tcaccaacaa gctcgtcatc
     6781 gctttgcaga agagcagaga ggatatgctc atcgtctaaa gaactaccca ttttattata
     6841 tattagtcac gatatctata acaagaaaat atatatataa taagttatca cgtaagtaga
     6901 acatgaaata acaatataat tatcgtatga gttaaatctt aaaagtcacg taaaagataa
     6961 tcatgcgtca ttttgactca cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat
     7021 tgacaagcac gcctcacggg agctccaagc ggcgactgag atgtcctaaa tgcacagcga
     7081 cggattcgcg ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac
     7141 gcagactatc tttctagggt taaaaaagat ttgcgcttta ctcgacctaa actttaaaca
     7201 cgtcatagaa tcttcgtttg acaaaaacca cattgtggcc aagctgtgtg acgcgacgcg
     7261 cgctaaagaa tggcaaacca agtcgcgcga gcgtcgactc tagaggatcc ccgggtaccg
     7321 agctcgaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7381 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7441 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7501 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7561 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7621 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7681 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7741 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7801 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7861 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7921 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7981 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     8041 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     8101 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     8161 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     8221 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     8281 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8341 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8401 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8461 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8521 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8581 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8641 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8701 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8761 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8821 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8881 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8941 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     9001 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     9061 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     9121 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     9181 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     9241 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9301 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9361 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9421 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9481 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     9541 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     9601 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     9661 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     9721 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     9781 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     9841 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     9901 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     9961 agctttgttt aaaaatataa caaaattgtg atcccacaaa atgaagtggg gcaaaatcaa
    10021 ataattaata gtgtccgtaa acttgttggt cttcaacttt ttgaggaaca cgttggacgg
    10081 caaatccgtg actataacac aagttgattt aataatttta gccaacacgt cgggctgcgt
    10141 gttttttgcc gacgcgtctg tgtacacgtt gattaactgg tcgattaaac tgttgaaata
    10201 atttaatttt tggttcttct ttaaatctgt gatgaaattt tttaaaataa ctttaaattc
    10261 ttcattggta aaaaatgcca cgttttgcaa cttgtgaggg tctaatatga ggtcaaactc
    10321 agtaggagtt ttatccaaaa aagaaaacat gattacgtct gtacacgaac gcgtattaac
    10381 gcagagtgca aagtataaga gggttaaaaa atatatttta cgcaccatat acgcatcggg
    10441 ttgatatcgt taatatggat caatttgaac agttgattaa cgtgtctctg ctcaagtctt
    10501 tgatcaaaac gcaaatcgac gaaaatgtgt cggacaatat caagtcgatg agcgaaaaac
    10561 taaaaaggct agaatacgac aatctcacag acagcgttga gatatacggt attcacgaca
    10621 gcaggctgaa taataaaaaa attagaaact attatttaac cctagaaaga taatcatatt
    10681 gtgacgtacg ttaaagataa tcatgcgtaa aattgacgca tgtgttttat cggtctgtat
    10741 atcgaggttt atttattaat ttgaatagat attaagtttt attatattta cacttacata
    10801 ctaataataa attcaacaaa caatttattt atgtttattt atttattaaa aaaaaacaaa
    10861 aactcaaaat ttcttctata aagtaacaaa acttttaaac attctctctt ttacaaaaat
    10921 aaacttattt tgtactttaa aaacagtcat gttgtattat aaaataagta attagcttaa
    10981 cttatacata atagaaacaa attatactta ttagtcagtc agaaacaact ttggcacata
    11041 tcaatattat gctctcgaca aataactttt ttgcattttt tgcacgatgc atttgccttt
    11101 cgccttattt tagaggggca gtaagtacag taagtacgtt ttttcattac tggctcttca
    11161 gtactgtcat ctgatgtacc aggcacttca tttggcaaaa tattagagat attatcgcgc
    11221 aaatatctct tcaaagtagg agcttctaaa cgcttacgca taaacgatga cgtcaggctc
    11281 atgtaaaggt ttctcataaa ttttttgcga ctttgaacct tttctccctt gctactgaca
    11341 ttatggctgt atataataaa agaatttatg caggcaatgt ttatcattcc gtacaataat
    11401 gccataggcc acctattcgt cttcctactg caggtcatca cagaacacat ttggtctagc
    11461 gtgtccactc cgcctttagt ttgattataa tacataacca tttgcggttt accggtactt
    11521 tcgttgatag aagcatcctc atcacaagat gataataagt ataccatctt agctggcttc
    11581 ggtttatatg agacgagagt aaggggtccg tcaaaacaaa acatcgatgt tcccactggc
    11641 ctggagcgac tgtttttcag tacttccggt atctcgcgtt tgtttgatcg cacggttccc
    11701 acaatggtta attcgagctc gcccggggtc ctaggtcgac gatgtaggtc acggtctcga
    11761 agccgcggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac tccacctcac
    11821 ccatctggtc catcatgatg aacgggtcga ggtggcggta gttgatcccg gcgaacgcgc
    11881 ggcgcaccgg gaagccctcg ccctcgaaac cgctgggcgc ggtggtcacg gtgagcacgg
    11941 gacgtgcgac ggcgtcggcg ggtgcggata cgcggggcag cgtcagcggg ttctcgacgg
    12001 tcacggcggg catgtcgacg cggccgcatg tgaatcaaca ttaggttgtc atgagaaaca
    12061 aacatgaatg tctaaatatt aaatcatgaa aatgaaaacc aactgatttg cttattaaat
    12121 tctaaatgta gtggctttag atagtcatgt gtgatagaac atgaatgtca tgagactgaa
    12181 ataagaaatt tgatagaaga aatcttgctc cccaaaatat ggattcgagg ctcctctgct
    12241 tgtcgcaagc tcacttgaat tttttttttt aaaacaaatg agtcagcaac aagcggagcg
    12301 ccgcaaatcc ataatttgtg aagtctggga ttagcatgtt tcattttatt cgaagctgaa
    12361 agctaggaaa atctgttagt ggaatccgat ttttctctaa caccatgcat tacatccaac
    12421 gctggaagta atgcatttta tttatagaaa attagaatga actcctcata ttcactcgga
    12481 catcttcact aatagaaaga aaaatgactc cgcaaatttt cctctaacac tttcagcttc
    12541 agaatttgcc tctttacttt ggaacatgat gatgactgtc gttcgacatg aggtccatcc
    12601 gcaatttagt tcgctatggg ttgatcacta cccaagtaac caataagctg tttatatcgg
    12661 caccaagtcg gttctatagt cgatttaaag gcttggtcga cagagccaat tggaactata
    12721 tcgtaataaa cggctgttat aaagccactt ttggcgaatt atttggctga taaacagcct
    12781 actcgtactg tgtctctgaa atgagtgatg tcaaaatttt atagcaagtg gcaaaaaaaa
    12841 gagaaaaaaa agttgtgtat cttctgttcg attttcattt cttttatgct ccctctggag
    12901 acgtttttta atttcaggct caaactgctg tcgtctaagt tcatattaat gattcccgtc
    12961 gcattttcta tctgctccat cgatgcacga gtggattggg atgcttaacg tcgtatttaa
    13021 aagagaaatg tttgcagcga tgtgtctatt tttgttctgc tgaatcacga gtgatgctgg
    13081 cgatgccggc aaggcatttg accatatatg tatagcagca caaaagaagc gaaatcaaca
    13141 gaaaaaaaat tacaatcaat ttacgaatct gttcacatct ttgagcatct taaattaaaa
    13201 cattcctgat aacgccgttt gttttgtttt ctgcggtgag agaaaaagag gcggagagat
    13261 gtgtttcgct ccattccctc gcatgttgac atttgcgtac gatatgaaat ctctttcgcc
    13321 gtaatcccga ttcagttttg ttttggttga taaacgtcat gacactaggc cgtttagaag
    13381 cattgacggt cgttgtgtac ggcttattgg cacagggctt acaagcacta aaataaagcg
    13441 ttatatactc atatacggcc aatatcagtg cctgatggtc acagtgccta taagcattcg
    13501 aaaatgtttt atgaagctag taggcactga aaatccgtta aatgtctcat ataccgctta
    13561 taacagtgct tgttggttac ttcttcttct tctttctggc gttacgtccc cactgggaca
    13621 gagcctgctt ctcggcttag tgttcttatg agcacttcca cagtaattat ctgagagctt
    13681 actatgccaa tgaccatttt tgcatgtgta tatcgtgtgg caggtacgaa gatactctat
    13741 gccctgggaa atcgagaaaa tttccaaccc gaaaatatcc tcgaccggtg ggattcgaac
    13801 ccacgaccct cagcttggtc ttgctgaata gctgcgcgtt taccgctacg gctatctggg
    13861 cccctgggta atgattcaga aatttggaac acggcaatcg acagcaagct catcgatttc
    13921 aaatcgacca acttgaatat gatgatcatg acacaagata atcataagtt aaacacactg
    13981 aatccgtgtt gggtgatgat ccatagcacc cataggttga cgcatacgaa tttgaaatgg
    14041 aaagctttag gaaatcatgt ggaaaactat ggaattcatg ttgtcggttg atgaatcagt
    14101 ccatgtagca aaactaaaga attgaatgtg taatacacat gaagtttgta agaaaatcac
    14161 aacaaatcga catagacgtt catgaatcga tccataattt taaacacacg gatcaagcgt
    14221 gtggattttt ttcagtgggg ggtgggtggg tgtcctcgtg gtgttacggc tcatacaaaa
    14281 tttgtaaact attcatataa aaagcgttac gagggggtgg gtgggtgtcg aaaatggcca
    14341 tttttggcgt tatgaaattt gtgaataaac ccatagttta tttcgaatac aatacttttc
    14401 actttcacag ccataaaaag gacgggctgc atttgaagtt tcatgacagt cgaatctgga
    14461 ctgcatgggc tcattcgcaa attttataac gctggagggg gtggatgggt gtcctagggt
    14521 tgttacagct catacaaaat tcgaaaaaca ttcatacaaa atgggttacg agggggtggg
    14581 taggtatgga aaatgaccat tttcggcgtt atgaaatttg ttgatatacc cagacgttga
    14641 tatatatttt tcctctgagc gagtctctct caggttccaa ccagcgaatc cccgctgtaa
    14701 caagcttcga ctgtactttg gccaccagcc gcgtaaaagc ttcatgaacg agctccttcg
    14761 ctctgatgca tgcgcagtca caggggctcc agctgtcaaa tcatccggca aaaccgaact
    14821 caaaacaaaa cgtgtttgtg tgtttcgtgt gtttccgcgc gttgctttct cttggataat
    14881 ttcacaaagt tttcgtgaat tgtgccgtaa aaatcgtgtt aattcaacag aatattttct
    14941 gtaattgttt gccagacgca cactgtttgc acgtaagtaa aaatccacaa agattgtcaa
    15001 aattctatca agtttttacc gcaaagtctt acgtccctca gaaggctcca ccctggggaa
    15061 actcgagatg
//
