LOCUS       sequence-193543-       15808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The promoter of AAEL007584 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 15808)
  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 15808)
  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: AAEL007584-Cas9
FEATURES             Location/Qualifiers
     source          1..15808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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)."
     primer_bind     598..620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   814..848
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    1516..1637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1527..1546)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1581..1600
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1751..2425,2426..2428,2429..2431))
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(1813..1836)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2011..2030)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2228..2248
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     CDS             complement(join(3666..4379,4380..4382,4383..4385))
                     /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(3711..3732)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4060..4079
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(4386..4439)
                     /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             complement(4440..4487)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4488..8588)
                     /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             complement(4488..8588)
                     /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             complement(8613..8633)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8640..8705)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    11956..12025
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     repeat_region   13059..13121
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(13825..13842)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(13825..13841)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13834..13856)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(14050..14069)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     14169..14191
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(14229..14247)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        14315..14419
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(14420..14488,14489..15280)
                     /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     complement(14638..14657)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(15451..15808,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gacatgatta cgaattcgag ctcggtaccc
      661 ggggatcctc tagagtcgac gctcgcgcga cttggtttgc cattctttag cgcgcgtcgc
      721 gtcacacagc ttggccacaa tgtggttttt gtcaaacgaa gattctatga cgtgtttaaa
      781 gtttaggtcg agtaaagcgc aaatcttttt taaccctaga aagatagtct gcgtaaaatt
      841 gacgcatgca ttcttgaaat attgctctct ctttctaaat agcgcgaatc cgtcgctgtg
      901 catttaggac atctcagtcg ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta
      961 agtgtcactg attttgaact ataacgaccg cgtgagtcaa aatgacgcat gattatcttt
     1021 tacgtgactt ttaagattta actcatacga taattatatt gttatttcat gttctactta
     1081 cgtgataact tattatatat atattttctt gttatagata tcgtgactaa tatataataa
     1141 aatgggtagt tctttagacg atgagcatat cctctctgct cttctgcaaa gcgatgacga
     1201 gcttgttggt gaggattctg acagtgaaat atcagatcac gtaagtgaag atgacgtcca
     1261 gagcgataca gaagaagcgt ttatagatga ggtacatgaa gtgcagccaa cgtcaagcgg
     1321 tagtgaaata ttagacgaac aaaatgttat tgaacaacca ggttcttcat tggcttctaa
     1381 cagaatcttg accttgccac agaggactat tagaggtaag aataaacatt gttggtcaac
     1441 ttcaaagtcc acgaggcgta gccgagtctc tgcactgaac attgtcagat ccgagatcgg
     1501 ccggcctaga agctttaaga tacattgatg agtttggaca aaccacaact agaatgcagt
     1561 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     1621 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1681 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     1741 atctagagtc ctacaggaac aggtggtggc ggccctcggt gcgctcgtac tgctccacga
     1801 tggtgtagtc ctcgttgtgg gaggtgatgt ccagcttgga gtccacgtag tagtagccgg
     1861 gcagctgcac gggcttcttg gccatgtaga tggacttgaa ctccaccagg tagtggccgc
     1921 cgtccttcag cttcagggcc ttgtggatct cgcccttcag cacgccgtcg cgggggtaca
     1981 ggcgctcggt ggaggcctcc cagcccatgg tcttcttctg cattacgggg ccgtcggagg
     2041 ggaagttcac gccgatgaac ttcaccttgt agatgaagca gccgtcctgc agggaggagt
     2101 cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc
     2161 cctcggggaa ggacagcttc ttgtagtcgg ggatgtcggc ggggtgcttc acgtacacct
     2221 tggagccgta ctggaactgg ggggacagga tgtcccaggc gaagggcagg gggccgccct
     2281 tggtcacctt cagcttcacg gtgttgtggc cctcgtaggg gcggccctcg ccctcgccct
     2341 cgatctcgaa ctcgtggccg ttcacggtgc cctccatgcg caccttgaag cgcatgaact
     2401 ccttgatgac gttcttggag gagcgcacca tcaccagaga caggttgcgg cggcggttgg
     2461 atggcgtggg cgcgttggcg ttgttggacc ggctcatgtt gtgtcgctgt aacagatgct
     2521 gttcaactgt gtttaccaga tcgttgcggg ctgtatttat aggcgcgata agcgggacgg
     2581 gcgcctcgtg tccggtcacg cgcatgagat aacgcgcggc tgatatggag gcgcgtcctg
     2641 ttccgataag gagttgcgtc cggctgcggt tagcaacaca ggaagctggc gtcctgtcac
     2701 gataagacaa cactcgtccg gtccgataat gtgattcgta cgtgacagga cgcgacccga
     2761 taaggccggc ctacgtgact gccgacacgt acttttttgc actgcaaaaa ggttcaatgt
     2821 gtggtagtgt atttggagcg tatacaacgg tgtagactat ttatgtaaaa tagtctacga
     2881 aacgtagagt ttgtactatg tatgggcccg cgtgcaaaag cgtgtttttt tgcagtgcaa
     2941 aaaagttggt ggtggggagg ccaccgagta tgggcgcgcc ccggccgtta actcgaatcg
     3001 ctatccaagc cagctccatt gtcggcatcg tgctcattct attgacgctg gcagatttgg
     3061 ttttggcgct atgggacccg ttcggttaca acaacatgtt tccgcgcgag tttcccgacg
     3121 acatgtcgcg cacgttcctg actgcgtact ttgagagttt cgacaacacc acgtccagag
     3181 aaatcataga gtttatgccc gagttctttt cggaaatggt cgaaacggac gatgacgcca
     3241 cgtttgaatc tctatttcat ttattagatt atgtggcatc tttagaagtt aattccgacg
     3301 gccaaatgtt aaacttggag gagggtgatg aaattgagga ttttgacgaa tctactttgg
     3361 tggggcaagc gttagccact agctcgctat acactcgcat ggagtttatg cagtacacgt
     3421 ttaggcaaaa cacactattg tctatgaaca aagaaaacaa caattttaat caaataatca
     3481 tgggtttatt tgcaacaaac acaattgtgg cgtttacagc atttgttata cacacagaac
     3541 tcatattttt tatatttttc gtaatcttcc taatgatcac attttattac ataatcaaag
     3601 aatcgtacga atattataaa acaattgatt tgttatttta aaaacgattc attctagtta
     3661 attaattact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     3721 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc
     3781 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     3841 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
     3901 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     3961 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     4021 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     4081 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     4141 tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc
     4201 acgagggtgg gccagggcac gggcagcttg ctggtggtgc agatgaactt cagggtcagc
     4261 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     4321 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     4381 accatagggc cgggattctc ctccacgtca ccgcatgtta gaagacttcc tctgccctcc
     4441 tttttctttt ttgcctggcc ggcctttttc gtggccgccg gccttttgtc gcctcccagc
     4501 tgagacaggt cgatccgtgt ctcgtacagg ccggtgatgc tctggtggat cagggtggcg
     4561 tccagcacct ctttggtgct ggtgtacctc ttccggtcga tggtggtgtc aaagtacttg
     4621 aaggcggcag gggctcccag attggtcagg gtaaacaggt ggatgatatt ctcggcctgc
     4681 tctctgatgg gcttatcccg gtgcttgttg taggcggaca gcactttgtc cagattagcg
     4741 tcggccagga tcactctctt ggagaactcg ctgatctgct cgatgatctc gtccaggtag
     4801 tgcttgtgct gttccacaaa cagctgtttc tgctcattat cctcggggga gcccttcagc
     4861 ttctcatagt ggctggccag gtacaggaag ttcacatatt tggagggcag ggccagttcg
     4921 tttcccttct gcagttcgcc ggcagaggcc agcattctct tccggccgtt ttccagctcg
     4981 aacagggagt acttaggcag cttgatgatc aggtcctttt tcacttcttt gtagcccttg
     5041 gcttccagaa agtcgatggg attcttctcg aagctgcttc tttccatgat ggtgatcccc
     5101 agcagctctt tcacactctt cagtttcttg gacttgccct tttccacttt ggccaccacc
     5161 agcacagaat aggccacggt ggggctgtcg aagccgccgt acttcttagg gtcccagtcc
     5221 ttctttctgg cgatcagctt atcgctgttc ctcttgggca ggatagactc tttgctgaag
     5281 ccgcctgtct gcacctcggt ctttttcacg atattcactt ggggcatgct cagcactttc
     5341 cgcacggtgg caaaatcccg gcccttatcc cacacgatct ccccggtttc gccgtttgtc
     5401 tcgatcagag gccgcttccg gatctcgccg ttggccaggg taatctcggt cttgaaaaag
     5461 ttcatgatgt tgctgtagaa gaagtacttg gcggtagcct tgccgatttc ctgctcgctc
     5521 ttggcgatca tcttccgcac gtcgtacacc ttgtagtcgc cgtacacgaa ctcgctttcc
     5581 agcttagggt actttttgat cagggcggtt cccacgacgg cgttcaggta ggcgtcgtgg
     5641 gcgtggtggt agttgttgat ctcgcgcact ttgtaaaact ggaaatcctt ccggaaatcg
     5701 gacaccagct tggacttcag ggtgatcact ttcacttccc ggatcagctt gtcattctcg
     5761 tcgtacttag tgttcatccg ggagtccagg atctgtgcca cgtgctttgt gatctgccgg
     5821 gtttccacca gctgtctctt gatgaagccg gccttatcca gttcgctcag gccgcctctc
     5881 tcggccttgg tcagattgtc gaactttctc tgggtaatca gcttggcgtt cagcagctgc
     5941 cgccagtagt tcttcatctt cttcacgacc tcttcggagg gcacgttgtc gctcttgccc
     6001 cggttcttgt cgcttctggt cagcaccttg ttgtcgatgg agtcgtcctt cagaaagctc
     6061 tgaggcacga tatggtccac atcgtagtcg gacagccggt tgatgtccag ttcctggtcc
     6121 acgtacatat cccgcccatt ctgcaggtag tacaggtaca gcttctcgtt ctgcagctgg
     6181 gtgttttcca cggggtgttc tttcaggatc tggctgccca gctctttgat gccctcttcg
     6241 atccgcttca ttctctcgcg gctgttcttc tgtcccttct gggtggtctg gttctctctg
     6301 gccatttcga tcacgatgtt ctcgggcttg tgccggccca tcactttcac gagctcgtcc
     6361 accaccttca ctgtctgcag gatgcccttc ttaatggcgg ggctgccggc cagattggca
     6421 atgtgctcgt gcaggctatc gccctggccg gacacctggg ctttctggat gtcctcttta
     6481 aaggtcaggc tgtcgtcgtg gatcagctgc atgaagtttc tgttggcgaa gccgtcggac
     6541 ttcaggaaat ccaggattgt cttgccggac tgcttgtccc ggatgccgtt gatcagcttc
     6601 cggctcagcc tgccccagcc ggtgtatctc cgccgcttca gctgcttcat cactttgtcg
     6661 tcgaacaggt gggcataggt tttcagccgt tcctcgatca tctctctgtc ctcaaacagt
     6721 gtcagggtca gcacgatatc ttccagaatg tcctcgtttt cctcattgtc caggaagtcc
     6781 ttgtccttga taattttcag cagatcgtgg tatgtgccca gggaggcgtt gaaccgatct
     6841 tccacgccgg agatttccac ggagtcgaag cactcgattt tcttgaagta gtcctctttc
     6901 agctgcttca cggtcacttt ccggttggtc ttgaacagca ggtccacgat ggcctttttc
     6961 tgctcgccgc tcaggaaggc gggctttctc attccctcgg tcacgtattt cactttggtc
     7021 agctcgttat acacggtgaa gtactcgtac agcaggctgt gcttgggcag caccttctcg
     7081 ttgggcaggt tcttatcgaa gttggtcatc cgctcgatga agctctgggc ggaagcgccc
     7141 ttgtccacca cttcctcgaa gttccagggg gtgatggttt cctcgctctt tctggtcatc
     7201 caggcgaatc tgctgtttcc cctggccaga gggcccacgt agtaggggat gcggaaggtc
     7261 aggatcttct cgatcttttc ccggttgtcc ttcaggaatg ggtaaaaatc ttcctgccgc
     7321 cgcagaatgg cgtgcagctc tcccaggtgg atctggtggg ggatgctgcc gttgtcgaag
     7381 gtccgctgct tccgcagcag gtcctctctg ttcagcttca cgagcagttc ctcggtgccg
     7441 tccatctttt ccaggatggg cttgatgaac ttgtagaact cttcctggct ggctccgccg
     7501 tcaatgtagc cggcgtagcc gttcttgctc tggtcgaaga aaatctcttt gtacttctca
     7561 ggcagctgct gccgcacgag agctttcagc agggtcaggt cctggtggtg ctcgtcgtat
     7621 ctcttgatca tagaggcgct caggggggcc ttggtgatct cggtgttcac tctcaggatg
     7681 tcgctcagca ggatggcgtc ggacaggttc ttggcggcca gaaacaggtc ggcgtactgg
     7741 tcgccgatct gggccagcag gttgtccagg tcgtcgtcgt aggtgtcctt gctcagctgc
     7801 agtttggcat cctcggccag gtcgaagttg ctcttgaagt tgggggtcag gcccaggctc
     7861 agggcaatca ggtttccgaa caggccattc ttcttctcgc cgggcagctg ggcgatcaga
     7921 ttttccagcc gtctgctctt gctcagtctg gcagacagga tggccttggc gtccacgccg
     7981 ctggcgttga tggggttttc ctcgaacagc tggttgtagg tctgcaccag ctggatgaac
     8041 agcttgtcca cgtcgctgtt gtcggggttc aggtcgccct cgatcaggaa gtggccccgg
     8101 aacttgatca tgtgggccag ggccagatag atcagccgca ggtcggcctt gtcggtgctg
     8161 tccaccagtt tctttctcag gtggtagatg gtggggtact tctcgtggta ggccacctcg
     8221 tccacgatgt tgccgaagat ggggtgccgc tcgtgcttct tatcctcttc caccaggaag
     8281 gactcttcca gtctgtggaa gaagctgtcg tccaccttgg ccatctcgtt gctgaagatc
     8341 tcttgcagat agcagatccg gttcttccgt ctggtgtatc ttcttctggc ggttctcttc
     8401 agccgggtgg cctcggctgt ttcgccgctg tcgaacagca gggctccgat caggttcttc
     8461 ttgatgctgt gccggtcggt gttgcccagc accttgaatt tcttgctggg caccttgtac
     8521 tcgtcggtga tcacggccca gcccacagag ttggtgccga tgtccaggcc gatgctgtac
     8581 ttcttgtcgg ctgctgggac tccgtggata ccgaccttcc gcttcttctt tggggccatc
     8641 ttatcgtcat cgtctttgta atcaatatca tgatccttgt agtctccgtc gtggtcctta
     8701 tagtccatct cgaggctgtg ataatgcgaa aaaacatttt ttgaatattt tggtagaaag
     8761 ataacttagt aagcactgtt tcattgcaat tccacttctg tttttgactc gactggagaa
     8821 ttccaaagct attcactcct tttattgaaa gactaggtat gaacgaaaca tagcaacggt
     8881 ctgctgtttt ggtatgaaat taggagaatc ataatttgta aggggaatcc atgtgatgtg
     8941 gtacctctga attgtttata ttttgtccaa aaataaaaga taatggttga tttcttttca
     9001 aagaacaaca atacggaaca ctacaaacga ctagtgaata gctgttgaaa tcttgtgtct
     9061 gtgcttggaa atcatttgaa acagtgctta aacatcttcc aaataattta aaaatattcc
     9121 acattctttt cggagcttac tgttttatac gaggttcact tcattcagtt gatccacaaa
     9181 tccgaccaaa aatcacccca aaacgatcga attctactga caaaccccgg atcaacaaac
     9241 gaatacttag taggctgttt caccactgaa ctggatggcc gtcttcaacg acaaccaaca
     9301 gctaccgttt ctaagtggtt cgaatgtgtg cgaacttcac agttgattac gtgttcacaa
     9361 tcaatatgtg gtgacatctc aattaggccc aacaatgaca caccagagcc tgccatccag
     9421 aaagcgggat ccatctgcgg tgggaaattg tcttgcaccg atgtccgtag tccatgtgag
     9481 gggtattaac ttttcaacaa ttgattccgt ttgcgtgcgt tgaaggagat gaaaaagaat
     9541 tttcttcgca ttggagatgg ttcgacaatg ggacggactc tataatccac gttaagagtc
     9601 ttatgtctcg gtcaccgcat cggaaacgtc cacgtggggg tggacgaagt aggcgcgcgc
     9661 ggtcgatggt agttctagca aggcaatttg tcacttgtca atggttgctt tgattaggat
     9721 tttgctctaa ggaaaggcgt tggaagtctg agaaaaaaat gcgtattctt tggaataaat
     9781 cactacaagg tgggatattc agtgctgcca ttttctttgc aaatgccgtt taaatattta
     9841 atttgttcca tgtatttagt catccatggg agaaactttt ggaagaatct caaatggatc
     9901 atttgacgaa atgcagctct acttctatac tcctctattt caccaaaaat gaaacaaagt
     9961 ccaccatgtt cctttcagtt gctcgaaatt ttatggcaca taagatagca aagaaaaata
    10021 caggataccg tgaaaagtgt attgcaatct gtcaaacttg tgtaaatacc tttagcagta
    10081 acaagggcca aaagttttac tagaagtatg gcacccactc tgagcccgtc tactatgtct
    10141 ttatttcgta tttgcttaca ctgctcattt gaatttacaa acattttatg ctcctggtaa
    10201 cgctaccatt tcaaatatac gagacatttt tcccgaacaa tatttaagtt gaatccctct
    10261 ttcacaattc acttaagttt ttgctgtatt ttgtcaagac ttctagtgaa acttttgagg
    10321 atgtctttgt gaaaccttta tgagcatgag catgattgac cgcccgaagt tgctactctg
    10381 ttattgcaag aacagctgta cttacacagg gaaccaacag atgctactca ggatcaatag
    10441 cattctcaat gtgtaagtac tggtgctctc attattatca caacgccggc tgcgtccgaa
    10501 tgcaggtcaa tttgggaatg ggagggaaat gttgacgtgt tacttgcttt acggaagccg
    10561 aggagtcctc tgcacttcca caagaaaaca ctgggagttt ggatatggaa aaggattcgt
    10621 ttaggtaaag ttataatttt taatgaatat gtgaacatat atacggatga tcgataccga
    10681 tagtgcggtt actaagtgaa ccggacacga ttcgaattcc gtcgctcgcc cacaaaggag
    10741 cgtgcaacag attcaacgga ccaaacacta ctcaatgtct ttgtgaaacc tttatggagg
    10801 attattacga gatttcctgt tgatatcact ttgaattttg ttggaaaatg ttctggaatg
    10861 tattgcggat aattttgtga attatttttt ggaaacattc gtgagacatt agtgaaactt
    10921 gtgtacatac ttcaatgaat aattatgtga aaattctatg gacacgtttg tggtgacttc
    10981 tgtggcacac tttgtaaaga cttttgtggt gctttatgtg aaaacttctt tggaatcctc
    11041 tgttaagagt tattttttta aagatatcct cgaagacttc tgttaagttt ctatggatgc
    11101 attcgtggaa acttctttgg gaaccttaat gaagacttct atggaagctt ttgtgggagc
    11161 tattgtgtag agttccacag agactttgta aatacttatt ggaagacata tgtggagatt
    11221 ttgtgattat tttcgtgaat aaatatttgg agcagaacat tcttgtggag agttttatag
    11281 aatcttctgt ggcaattttg tgggacactt tggtgaatat agtatatgta cttctgtaat
    11341 aaaaatctgt ggacacctct gcagatgcct gtttcgatga agatttctgt agagatgaaa
    11401 gaaaaaacaa gacgatcata gaaattgcaa cagcggtgtc taagtcgtct actttgattc
    11461 ccaagaggta attgtaaagt ctccaagagg ttctcaaggg gttttcggag gttcaccaaa
    11521 agttccctga ggtttcctaa acatttatta gttttacagt atttttaggg attccgacca
    11581 tcctttaaaa aaaatctata aaggaacatc tgaaattttt aaagattttt cggctttttg
    11641 ttaacagaat aacggcaaaa cattcacgaa actggttatt gcacacaaat tacaaaattt
    11701 aaattccact tttatggctg gtacgctgat cataagtacc gtatttcggg tgagcggccg
    11761 cgtcgacatg cccgccgtga ccgtcgagaa cccgctgacg ctgccccgcg tatccgcacc
    11821 cgccgacgcc gtcgcacgtc ccgtgctcac cgtgaccacc gcgcccagcg gtttcgaggg
    11881 cgagggcttc ccggtgcgcc gcgcgttcgc cgggatcaac taccgccacc tcgacccgtt
    11941 catcatgatg gaccagatgg gtgaggtgga gtacgcgccc ggggagccca agggcacgcc
    12001 ctggcacccg caccgcggct tcgagaccgt gacctacatc gtcgacctag gaccccgggc
    12061 gagctcgaat taaccattgt gggaaccgtg cgatcaaaca aacgcgagat accggaagta
    12121 ctgaaaaaca gtcgctccag gccagtggga acatcgatgt tttgttttga cggacccctt
    12181 actctcgtct catataaacc gaagccagct aagatggtat acttattatc atcttgtgat
    12241 gaggatgctt ctatcaacga aagtaccggt aaaccgcaaa tggttatgta ttataatcaa
    12301 actaaaggcg gagtggacac gctagaccaa atgtgttctg tgatgacctg cagtaggaag
    12361 acgaataggt ggcctatggc attattgtac ggaatgataa acattgcctg cataaattct
    12421 tttattatat acagccataa tgtcagtagc aagggagaaa aggttcaaag tcgcaaaaaa
    12481 tttatgagaa acctttacat gagcctgacg tcatcgttta tgcgtaagcg tttagaagct
    12541 cctactttga agagatattt gcgcgataat atctctaata ttttgccaaa tgaagtgcct
    12601 ggtacatcag atgacagtac tgaagagcca gtaatgaaaa aacgtactta ctgtacttac
    12661 tgcccctcta aaataaggcg aaaggcaaat gcatcgtgca aaaaatgcaa aaaagttatt
    12721 tgtcgagagc ataatattga tatgtgccaa agttgtttct gactgactaa taagtataat
    12781 ttgtttctat tatgtataag ttaagctaat tacttatttt ataatacaac atgactgttt
    12841 ttaaagtaca aaataagttt atttttgtaa aagagagaat gtttaaaagt tttgttactt
    12901 tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat aaataaattg
    12961 tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat atctattcaa
    13021 attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt ttacgcatga
    13081 ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaaataat agtttctaat
    13141 ttttttatta ttcagcctgc tgtcgtgaat accgtatatc tcaacgctgt ctgtgagatt
    13201 gtcgtattct agccttttta gtttttcgct catcgacttg atattgtccg acacattttc
    13261 gtcgatttgc gttttgatca aagacttgag cagagacacg ttaatcaact gttcaaattg
    13321 atccatatta acgatatcaa cccgatgcgt atatggtgcg taaaatatat tttttaaccc
    13381 tcttatactt tgcactctgc gttaatacgc gttcgtgtac agacgtaatc atgttttctt
    13441 ttttggataa aactcctact gagtttgacc tcatattaga ccctcacaag ttgcaaaacg
    13501 tggcattttt taccaatgaa gaatttaaag ttattttaaa aaatttcatc acagatttaa
    13561 agaagaacca aaaattaaat tatttcaaca gtttaatcga ccagttaatc aacgtgtaca
    13621 cagacgcgtc ggcaaaaaac acgcagcccg acgtgttggc taaaattatt aaatcaactt
    13681 gtgttatagt cacggatttg ccgtccaacg tgttcctcaa aaagttgaag accaacaagt
    13741 ttacggacac tattaattat ttgattttgc cccacttcat tttgtgggat cacaattttg
    13801 ttatattttt aaacaaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
    13861 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
    13921 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
    13981 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
    14041 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
    14101 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
    14161 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
    14221 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
    14281 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
    14341 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
    14401 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
    14461 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
    14521 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
    14581 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
    14641 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
    14701 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
    14761 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
    14821 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
    14881 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
    14941 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
    15001 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
    15061 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
    15121 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
    15181 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
    15241 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
    15301 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
    15361 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
    15421 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
    15481 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
    15541 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
    15601 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
    15661 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
    15721 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
    15781 gtgcacacag cccagcttgg agcgaacg
//
