LOCUS       sequence-193541-       15803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The promoter of AAEL005635 drive expression of Streptococcus 
            pyogenes Cas9 gene in Aedes aegypti.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15803)
  AUTHORS   Li M, Bui M, Yang T, Bowman CS, White BJ, Akbari OS
  TITLE     Germline Cas9 expression yields highly efficient genome engineering 
            in a major worldwide disease vector, Aedes aegypti.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Nov 14. pii: 201711538. doi: 
            10.1073/pnas.1711538114.
  PUBMED    29138316
REFERENCE   2  (bases 1 to 15803)
  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: "Proc Natl 
            Acad Sci U S A. 2017 Nov 14. pii: 201711538. doi: 
            10.1073/pnas.1711538114."
COMMENT     Sequence Label: AAEL005635-Cas9
FEATURES             Location/Qualifiers
     source          1..15803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             905..970
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             977..997
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1022..5122
                     /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             1022..5122
                     /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             5123..5170
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5171..5224
                     /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(5225..5227,5228..5230,5231..5944)
                     /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(5531..5550)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5878..5899
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(7179..7181,7182..7184,7185..7859)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(7362..7382)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     7580..7599
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7774..7797
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    7973..8094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8010..8029)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8064..8083
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8762..8796
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8977..8993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8977..8993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8990..9012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9001..9017
                     /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(9025..9031,9032..9049,9050..9055))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9070..9091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9208..9225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9379..9967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9459..9478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10138..10929,10930..10998))
                     /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     10761..10780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10999..11103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     11171..11189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(11227..11249)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     11349..11368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11562..11584
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11576..11593
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11577..11593
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   12297..12359
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    13393..13462
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
ORIGIN
        1 tcgacataga cgttcatgaa tcgatccata attttaaaca cacggatcaa gcgtgtggat
       61 ttttttcagt ggggggtggg tgggtgtcct cgtggtgtta cggctcatac aaaatttgta
      121 aactattcat ataaaaagcg ttacgagggg gtgggtgggt gtcgaaaatg gccatttttg
      181 gcgttatgaa atttgtgaat aaacccatag tttatttcga atacaatact tttcactttc
      241 acagccataa aaaggacggg ctgcatttga agtttcatga cagtcgaatc tggactgcat
      301 gggctcattc gcaaatttta taacgctgga gggggtggat gggtgtccta gggttgttac
      361 agctcataca aaattcgaaa aacattcata caaaatgggt tacgaggggg tgggtaggta
      421 tggaaaatga ccattttcgg cgttatgaaa tttgttgata tacccagacg ttgatatata
      481 tttttcctct gagcgagtct ctctcaggtt ccaaccagcg aatccccgct gtaacaagct
      541 tcgactgtac tttggccacc agccgcgtaa aagcttcatg aacgagctcc ttcgctctga
      601 tgcatgcgca gtcacagggg ctccagctgt caaatcatcc ggcaaaaccg aactcaaaac
      661 aaaacgtgtt tgtgtgtttc gtgtgtttcc gcgcgttgct ttctcttgga taatttcaca
      721 aagttttcgt gaattgtgcc gtaaaaatcg tgttaattca acagaatatt ttctgtaatt
      781 gtttgccaga cgcacactgt ttgcacgtaa gtaaaaatcc acaaagattg tcaaaattct
      841 atcaagtttt taccgcaaag tcttacgtcc ctcagaaggc tccaccctgg ggaaactcga
      901 gatggactat aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga
      961 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
     1021 cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat
     1081 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
     1141 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
     1201 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
     1261 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
     1321 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
     1381 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
     1441 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
     1501 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
     1561 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
     1621 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
     1681 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
     1741 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
     1801 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
     1861 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     1921 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     1981 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     2041 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     2101 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     2161 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     2221 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     2281 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     2341 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     2401 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     2461 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     2521 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     2581 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     2641 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     2701 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     2761 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     2821 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     2881 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     2941 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     3001 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     3061 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     3121 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     3181 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     3241 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     3301 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     3361 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     3421 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     3481 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
     3541 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
     3601 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     3661 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     3721 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     3781 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     3841 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     3901 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     3961 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     4021 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     4081 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     4141 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
     4201 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     4261 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     4321 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     4381 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     4441 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     4501 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     4561 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     4621 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     4681 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     4741 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     4801 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     4861 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     4921 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     4981 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     5041 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     5101 cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
     5161 aaagaaaaag gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg
     5221 ccctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     5281 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     5341 ctacggcaag ctgaccctga agttcatctg caccaccagc aagctgcccg tgccctggcc
     5401 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     5461 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     5521 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     5581 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     5641 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     5701 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     5761 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     5821 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     5881 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     5941 gtaattaatt aactagaatg aatcgttttt aaaataacaa atcaattgtt ttataatatt
     6001 cgtacgattc tttgattatg taataaaatg tgatcattag gaagattacg aaaaatataa
     6061 aaaatatgag ttctgtgtgt ataacaaatg ctgtaaacgc cacaattgtg tttgttgcaa
     6121 ataaacccat gattatttga ttaaaattgt tgttttcttt gttcatagac aatagtgtgt
     6181 tttgcctaaa cgtgtactgc ataaactcca tgcgagtgta tagcgagcta gtggctaacg
     6241 cttgccccac caaagtagat tcgtcaaaat cctcaatttc atcaccctcc tccaagttta
     6301 acatttggcc gtcggaatta acttctaaag atgccacata atctaataaa tgaaatagag
     6361 attcaaacgt ggcgtcatcg tccgtttcga ccatttccga aaagaactcg ggcataaact
     6421 ctatgatttc tctggacgtg gtgttgtcga aactctcaaa gtacgcagtc aggaacgtgc
     6481 gcgacatgtc gtcgggaaac tcgcgcggaa acatgttgtt gtaaccgaac gggtcccata
     6541 gcgccaaaac caaatctgcc agcgtcaata gaatgagcac gatgccgaca atggagctgg
     6601 cttggatagc gattcgagtt aacggccggg gcgcgcccat actcggtggc ctccccacca
     6661 ccaacttttt tgcactgcaa aaaaacacgc ttttgcacgc gggcccatac atagtacaaa
     6721 ctctacgttt cgtagactat tttacataaa tagtctacac cgttgtatac gctccaaata
     6781 cactaccaca cattgaacct ttttgcagtg caaaaaagta cgtgtcggca gtcacgtagg
     6841 ccggccttat cgggtcgcgt cctgtcacgt acgaatcaca ttatcggacc ggacgagtgt
     6901 tgtcttatcg tgacaggacg ccagcttcct gtgttgctaa ccgcagccgg acgcaactcc
     6961 ttatcggaac aggacgcgcc tccatatcag ccgcgcgtta tctcatgcgc gtgaccggac
     7021 acgaggcgcc cgtcccgctt atcgcgccta taaatacagc ccgcaacgat ctggtaaaca
     7081 cagttgaaca gcatctgtta cagcgacaca acatgagccg gtccaacaac gccaacgcgc
     7141 ccacgccatc caaccgccgc cgcaacctgt ctctggtgat ggtgcgctcc tccaagaacg
     7201 tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg caccgtgaac ggccacgagt
     7261 tcgagatcga gggcgagggc gagggccgcc cctacgaggg ccacaacacc gtgaagctga
     7321 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttccagt
     7381 acggctccaa ggtgtacgtg aagcaccccg ccgacatccc cgactacaag aagctgtcct
     7441 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     7501 tgacccagga ctcctccctg caggacggct gcttcatcta caaggtgaag ttcatcggcg
     7561 tgaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca
     7621 ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccacaag gccctgaagc
     7681 tgaaggacgg cggccactac ctggtggagt tcaagtccat ctacatggcc aagaagcccg
     7741 tgcagctgcc cggctactac tacgtggact ccaagctgga catcacctcc cacaacgagg
     7801 actacaccat cgtggagcag tacgagcgca ccgagggccg ccaccacctg ttcctgtagg
     7861 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     7921 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     7981 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     8041 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaagctt
     8101 ctaggccggc cgatctcgga tctgacaatg ttcagtgcag agactcggct acgcctcgtg
     8161 gactttgaag ttgaccaaca atgtttattc ttacctctaa tagtcctctg tggcaaggtc
     8221 aagattctgt tagaagccaa tgaagaacct ggttgttcaa taacattttg ttcgtctaat
     8281 atttcactac cgcttgacgt tggctgcact tcatgtacct catctataaa cgcttcttct
     8341 gtatcgctct ggacgtcatc ttcacttacg tgatctgata tttcactgtc agaatcctca
     8401 ccaacaagct cgtcatcgct ttgcagaaga gcagagagga tatgctcatc gtctaaagaa
     8461 ctacccattt tattatatat tagtcacgat atctataaca agaaaatata tatataataa
     8521 gttatcacgt aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa
     8581 agtcacgtaa aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc
     8641 agtgacactt accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg
     8701 tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat
     8761 gcatgcgtca attttacgca gactatcttt ctagggttaa aaaagatttg cgctttactc
     8821 gacctaaact ttaaacacgt catagaatct tcgtttgaca aaaaccacat tgtggccaag
     8881 ctgtgtgacg cgacgcgcgc taaagaatgg caaaccaagt cgcgcgagcg tcgactctag
     8941 aggatccccg ggtaccgagc tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     9001 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     9061 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     9121 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     9181 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     9241 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     9301 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     9361 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     9421 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     9481 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     9541 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     9601 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     9661 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     9721 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     9781 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     9841 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     9901 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     9961 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
    10021 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
    10081 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
    10141 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
    10201 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
    10261 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
    10321 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
    10381 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
    10441 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
    10501 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
    10561 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
    10621 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
    10681 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
    10741 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
    10801 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
    10861 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
    10921 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
    10981 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
    11041 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
    11101 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
    11161 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
    11221 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
    11281 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
    11341 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
    11401 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
    11461 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
    11521 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
    11581 aacgacggcc agtgccaagc tttgtttaaa aatataacaa aattgtgatc ccacaaaatg
    11641 aagtggggca aaatcaaata attaatagtg tccgtaaact tgttggtctt caactttttg
    11701 aggaacacgt tggacggcaa atccgtgact ataacacaag ttgatttaat aattttagcc
    11761 aacacgtcgg gctgcgtgtt ttttgccgac gcgtctgtgt acacgttgat taactggtcg
    11821 attaaactgt tgaaataatt taatttttgg ttcttcttta aatctgtgat gaaatttttt
    11881 aaaataactt taaattcttc attggtaaaa aatgccacgt tttgcaactt gtgagggtct
    11941 aatatgaggt caaactcagt aggagtttta tccaaaaaag aaaacatgat tacgtctgta
    12001 cacgaacgcg tattaacgca gagtgcaaag tataagaggg ttaaaaaata tattttacgc
    12061 accatatacg catcgggttg atatcgttaa tatggatcaa tttgaacagt tgattaacgt
    12121 gtctctgctc aagtctttga tcaaaacgca aatcgacgaa aatgtgtcgg acaatatcaa
    12181 gtcgatgagc gaaaaactaa aaaggctaga atacgacaat ctcacagaca gcgttgagat
    12241 atacggtatt cacgacagca ggctgaataa taaaaaaatt agaaactatt atttaaccct
    12301 agaaagataa tcatattgtg acgtacgtta aagataatca tgcgtaaaat tgacgcatgt
    12361 gttttatcgg tctgtatatc gaggtttatt tattaatttg aatagatatt aagttttatt
    12421 atatttacac ttacatacta ataataaatt caacaaacaa tttatttatg tttatttatt
    12481 tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact tttaaacatt
    12541 ctctctttta caaaaataaa cttattttgt actttaaaaa cagtcatgtt gtattataaa
    12601 ataagtaatt agcttaactt atacataata gaaacaaatt atacttatta gtcagtcaga
    12661 aacaactttg gcacatatca atattatgct ctcgacaaat aacttttttg cattttttgc
    12721 acgatgcatt tgcctttcgc cttattttag aggggcagta agtacagtaa gtacgttttt
    12781 tcattactgg ctcttcagta ctgtcatctg atgtaccagg cacttcattt ggcaaaatat
    12841 tagagatatt atcgcgcaaa tatctcttca aagtaggagc ttctaaacgc ttacgcataa
    12901 acgatgacgt caggctcatg taaaggtttc tcataaattt tttgcgactt tgaacctttt
    12961 ctcccttgct actgacatta tggctgtata taataaaaga atttatgcag gcaatgttta
    13021 tcattccgta caataatgcc ataggccacc tattcgtctt cctactgcag gtcatcacag
    13081 aacacatttg gtctagcgtg tccactccgc ctttagtttg attataatac ataaccattt
    13141 gcggtttacc ggtactttcg ttgatagaag catcctcatc acaagatgat aataagtata
    13201 ccatcttagc tggcttcggt ttatatgaga cgagagtaag gggtccgtca aaacaaaaca
    13261 tcgatgttcc cactggcctg gagcgactgt ttttcagtac ttccggtatc tcgcgtttgt
    13321 ttgatcgcac ggttcccaca atggttaatt cgagctcgcc cggggtccta ggtcgacgat
    13381 gtaggtcacg gtctcgaagc cgcggtgcgg gtgccagggc gtgcccttgg gctccccggg
    13441 cgcgtactcc acctcaccca tctggtccat catgatgaac gggtcgaggt ggcggtagtt
    13501 gatcccggcg aacgcgcggc gcaccgggaa gccctcgccc tcgaaaccgc tgggcgcggt
    13561 ggtcacggtg agcacgggac gtgcgacggc gtcggcgggt gcggatacgc ggggcagcgt
    13621 cagcgggttc tcgacggtca cggcgggcat gtcgacgcgg ccgcatgtga atcaacatta
    13681 ggttgtcatg agaaacaaac atgaatgtct aaatattaaa tcatgaaaat gaaaaccaac
    13741 tgatttgctt attaaattct aaatgtagtg gctttagata gtcatgtgtg atagaacatg
    13801 aatgtcatga gactgaaata agaaatttga tagaagaaat cttgctcccc aaaatatgga
    13861 ttcgaggctc ctctgcttgt cgcaagctca cttgaatttt tttttttaaa acaaatgagt
    13921 cagcaacaag cggagcgccg caaatccata atttgtgaag tctgggatta gcatgtttca
    13981 ttttattcga agctgaaagc taggaaaatc tgttagtgga atccgatttt tctctaacac
    14041 catgcattac atccaacgct ggaagtaatg cattttattt atagaaaatt agaatgaact
    14101 cctcatattc actcggacat cttcactaat agaaagaaaa atgactccgc aaattttcct
    14161 ctaacacttt cagcttcaga atttgcctct ttactttgga acatgatgat gactgtcgtt
    14221 cgacatgagg tccatccgca atttagttcg ctatgggttg atcactaccc aagtaaccaa
    14281 taagctgttt atatcggcac caagtcggtt ctatagtcga tttaaaggct tggtcgacag
    14341 agccaattgg aactatatcg taataaacgg ctgttataaa gccacttttg gcgaattatt
    14401 tggctgataa acagcctact cgtactgtgt ctctgaaatg agtgatgtca aaattttata
    14461 gcaagtggca aaaaaaagag aaaaaaaagt tgtgtatctt ctgttcgatt ttcatttctt
    14521 ttatgctccc tctggagacg ttttttaatt tcaggctcaa actgctgtcg tctaagttca
    14581 tattaatgat tcccgtcgca ttttctatct gctccatcga tgcacgagtg gattgggatg
    14641 cttaacgtcg tatttaaaag agaaatgttt gcagcgatgt gtctattttt gttctgctga
    14701 atcacgagtg atgctggcga tgccggcaag gcatttgacc atatatgtat agcagcacaa
    14761 aagaagcgaa atcaacagaa aaaaaattac aatcaattta cgaatctgtt cacatctttg
    14821 agcatcttaa attaaaacat tcctgataac gccgtttgtt ttgttttctg cggtgagaga
    14881 aaaagaggcg gagagatgtg tttcgctcca ttccctcgca tgttgacatt tgcgtacgat
    14941 atgaaatctc tttcgccgta atcccgattc agttttgttt tggttgataa acgtcatgac
    15001 actaggccgt ttagaagcat tgacggtcgt tgtgtacggc ttattggcac agggcttaca
    15061 agcactaaaa taaagcgtta tatactcata tacggccaat atcagtgcct gatggtcaca
    15121 gtgcctataa gcattcgaaa atgttttatg aagctagtag gcactgaaaa tccgttaaat
    15181 gtctcatata ccgcttataa cagtgcttgt tggttacttc ttcttcttct ttctggcgtt
    15241 acgtccccac tgggacagag cctgcttctc ggcttagtgt tcttatgagc acttccacag
    15301 taattatctg agagcttact atgccaatga ccatttttgc atgtgtatat cgtgtggcag
    15361 gtacgaagat actctatgcc ctgggaaatc gagaaaattt ccaacccgaa aatatcctcg
    15421 accggtggga ttcgaaccca cgaccctcag cttggtcttg ctgaatagct gcgcgtttac
    15481 cgctacggct atctgggccc ctgggtaatg attcagaaat ttggaacacg gcaatcgaca
    15541 gcaagctcat cgatttcaaa tcgaccaact tgaatatgat gatcatgaca caagataatc
    15601 ataagttaaa cacactgaat ccgtgttggg tgatgatcca tagcacccat aggttgacgc
    15661 atacgaattt gaaatggaaa gctttaggaa atcatgtgga aaactatgga attcatgttg
    15721 tcggttgatg aatcagtcca tgtagcaaaa ctaaagaatt gaatgtgtaa tacacatgaa
    15781 gtttgtaaga aaatcacaac aaa
//
