LOCUS       sequence-193542-       15271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The promoter of AAEL007097 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 15271)
  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 15271)
  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: AAEL007097-Cas9
FEATURES             Location/Qualifiers
     source          1..15271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(460..513)
                     /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(514..561)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(562..4662)
                     /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(562..4662)
                     /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(4687..4707)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4714..4779)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    7493..7562
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     repeat_region   8596..8658
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9362..9379)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9362..9378)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9371..9393)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(9587..9606)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9706..9728
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9766..9784)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9852..9956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9957..10025,10026..10817)
                     /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(10175..10194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10988..11576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11477..11496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11730..11747
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11864..11885
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11900..11905,11906..11923,11924..11930)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11938..11954
                     /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     11943..11965
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11962..11978
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11962..11978
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   12159..12193
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    12861..12982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12872..12891)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     12926..12945
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(13096..13770,13771..13773,13774..13776))
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(13158..13181)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(13356..13375)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     13573..13593
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     CDS             complement(join(15011..15271,1..453,454..456,457..459))
                     /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(15056..15077)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
       61 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
      121 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
      181 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
      241 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
      301 cttgctggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
      361 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
      421 caccaccccg gtgaacagct cctcgccctt gctcaccata gggccgggat tctcctccac
      481 gtcaccgcat gttagaagac ttcctctgcc ctcctttttc ttttttgcct ggccggcctt
      541 tttcgtggcc gccggccttt tgtcgcctcc cagctgagac aggtcgatcc gtgtctcgta
      601 caggccggtg atgctctggt ggatcagggt ggcgtccagc acctctttgg tgctggtgta
      661 cctcttccgg tcgatggtgg tgtcaaagta cttgaaggcg gcaggggctc ccagattggt
      721 cagggtaaac aggtggatga tattctcggc ctgctctctg atgggcttat cccggtgctt
      781 gttgtaggcg gacagcactt tgtccagatt agcgtcggcc aggatcactc tcttggagaa
      841 ctcgctgatc tgctcgatga tctcgtccag gtagtgcttg tgctgttcca caaacagctg
      901 tttctgctca ttatcctcgg gggagccctt cagcttctca tagtggctgg ccaggtacag
      961 gaagttcaca tatttggagg gcagggccag ttcgtttccc ttctgcagtt cgccggcaga
     1021 ggccagcatt ctcttccggc cgttttccag ctcgaacagg gagtacttag gcagcttgat
     1081 gatcaggtcc tttttcactt ctttgtagcc cttggcttcc agaaagtcga tgggattctt
     1141 ctcgaagctg cttctttcca tgatggtgat ccccagcagc tctttcacac tcttcagttt
     1201 cttggacttg cccttttcca ctttggccac caccagcaca gaataggcca cggtggggct
     1261 gtcgaagccg ccgtacttct tagggtccca gtccttcttt ctggcgatca gcttatcgct
     1321 gttcctcttg ggcaggatag actctttgct gaagccgcct gtctgcacct cggtcttttt
     1381 cacgatattc acttggggca tgctcagcac tttccgcacg gtggcaaaat cccggccctt
     1441 atcccacacg atctccccgg tttcgccgtt tgtctcgatc agaggccgct tccggatctc
     1501 gccgttggcc agggtaatct cggtcttgaa aaagttcatg atgttgctgt agaagaagta
     1561 cttggcggta gccttgccga tttcctgctc gctcttggcg atcatcttcc gcacgtcgta
     1621 caccttgtag tcgccgtaca cgaactcgct ttccagctta gggtactttt tgatcagggc
     1681 ggttcccacg acggcgttca ggtaggcgtc gtgggcgtgg tggtagttgt tgatctcgcg
     1741 cactttgtaa aactggaaat ccttccggaa atcggacacc agcttggact tcagggtgat
     1801 cactttcact tcccggatca gcttgtcatt ctcgtcgtac ttagtgttca tccgggagtc
     1861 caggatctgt gccacgtgct ttgtgatctg ccgggtttcc accagctgtc tcttgatgaa
     1921 gccggcctta tccagttcgc tcaggccgcc tctctcggcc ttggtcagat tgtcgaactt
     1981 tctctgggta atcagcttgg cgttcagcag ctgccgccag tagttcttca tcttcttcac
     2041 gacctcttcg gagggcacgt tgtcgctctt gccccggttc ttgtcgcttc tggtcagcac
     2101 cttgttgtcg atggagtcgt ccttcagaaa gctctgaggc acgatatggt ccacatcgta
     2161 gtcggacagc cggttgatgt ccagttcctg gtccacgtac atatcccgcc cattctgcag
     2221 gtagtacagg tacagcttct cgttctgcag ctgggtgttt tccacggggt gttctttcag
     2281 gatctggctg cccagctctt tgatgccctc ttcgatccgc ttcattctct cgcggctgtt
     2341 cttctgtccc ttctgggtgg tctggttctc tctggccatt tcgatcacga tgttctcggg
     2401 cttgtgccgg cccatcactt tcacgagctc gtccaccacc ttcactgtct gcaggatgcc
     2461 cttcttaatg gcggggctgc cggccagatt ggcaatgtgc tcgtgcaggc tatcgccctg
     2521 gccggacacc tgggctttct ggatgtcctc tttaaaggtc aggctgtcgt cgtggatcag
     2581 ctgcatgaag tttctgttgg cgaagccgtc ggacttcagg aaatccagga ttgtcttgcc
     2641 ggactgcttg tcccggatgc cgttgatcag cttccggctc agcctgcccc agccggtgta
     2701 tctccgccgc ttcagctgct tcatcacttt gtcgtcgaac aggtgggcat aggttttcag
     2761 ccgttcctcg atcatctctc tgtcctcaaa cagtgtcagg gtcagcacga tatcttccag
     2821 aatgtcctcg ttttcctcat tgtccaggaa gtccttgtcc ttgataattt tcagcagatc
     2881 gtggtatgtg cccagggagg cgttgaaccg atcttccacg ccggagattt ccacggagtc
     2941 gaagcactcg attttcttga agtagtcctc tttcagctgc ttcacggtca ctttccggtt
     3001 ggtcttgaac agcaggtcca cgatggcctt tttctgctcg ccgctcagga aggcgggctt
     3061 tctcattccc tcggtcacgt atttcacttt ggtcagctcg ttatacacgg tgaagtactc
     3121 gtacagcagg ctgtgcttgg gcagcacctt ctcgttgggc aggttcttat cgaagttggt
     3181 catccgctcg atgaagctct gggcggaagc gcccttgtcc accacttcct cgaagttcca
     3241 gggggtgatg gtttcctcgc tctttctggt catccaggcg aatctgctgt ttcccctggc
     3301 cagagggccc acgtagtagg ggatgcggaa ggtcaggatc ttctcgatct tttcccggtt
     3361 gtccttcagg aatgggtaaa aatcttcctg ccgccgcaga atggcgtgca gctctcccag
     3421 gtggatctgg tgggggatgc tgccgttgtc gaaggtccgc tgcttccgca gcaggtcctc
     3481 tctgttcagc ttcacgagca gttcctcggt gccgtccatc ttttccagga tgggcttgat
     3541 gaacttgtag aactcttcct ggctggctcc gccgtcaatg tagccggcgt agccgttctt
     3601 gctctggtcg aagaaaatct ctttgtactt ctcaggcagc tgctgccgca cgagagcttt
     3661 cagcagggtc aggtcctggt ggtgctcgtc gtatctcttg atcatagagg cgctcagggg
     3721 ggccttggtg atctcggtgt tcactctcag gatgtcgctc agcaggatgg cgtcggacag
     3781 gttcttggcg gccagaaaca ggtcggcgta ctggtcgccg atctgggcca gcaggttgtc
     3841 caggtcgtcg tcgtaggtgt ccttgctcag ctgcagtttg gcatcctcgg ccaggtcgaa
     3901 gttgctcttg aagttggggg tcaggcccag gctcagggca atcaggtttc cgaacaggcc
     3961 attcttcttc tcgccgggca gctgggcgat cagattttcc agccgtctgc tcttgctcag
     4021 tctggcagac aggatggcct tggcgtccac gccgctggcg ttgatggggt tttcctcgaa
     4081 cagctggttg taggtctgca ccagctggat gaacagcttg tccacgtcgc tgttgtcggg
     4141 gttcaggtcg ccctcgatca ggaagtggcc ccggaacttg atcatgtggg ccagggccag
     4201 atagatcagc cgcaggtcgg ccttgtcggt gctgtccacc agtttctttc tcaggtggta
     4261 gatggtgggg tacttctcgt ggtaggccac ctcgtccacg atgttgccga agatggggtg
     4321 ccgctcgtgc ttcttatcct cttccaccag gaaggactct tccagtctgt ggaagaagct
     4381 gtcgtccacc ttggccatct cgttgctgaa gatctcttgc agatagcaga tccggttctt
     4441 ccgtctggtg tatcttcttc tggcggttct cttcagccgg gtggcctcgg ctgtttcgcc
     4501 gctgtcgaac agcagggctc cgatcaggtt cttcttgatg ctgtgccggt cggtgttgcc
     4561 cagcaccttg aatttcttgc tgggcacctt gtactcgtcg gtgatcacgg cccagcccac
     4621 agagttggtg ccgatgtcca ggccgatgct gtacttcttg tcggctgctg ggactccgtg
     4681 gataccgacc ttccgcttct tctttggggc catcttatcg tcatcgtctt tgtaatcaat
     4741 atcatgatcc ttgtagtctc cgtcgtggtc cttatagtcc atgtttaaac ttggattttg
     4801 gacacttatt aacacactta tttaaaacaa ctattgaata cacttagttt tctgtcactt
     4861 tgtcacttga atataaaact taactttctt ctctggatag tagttcaatt ttgaagcggt
     4921 gagtgacctc aaggccctat ttatagtcga gagagataaa ggagatctgc atggttagcc
     4981 catgttgcgg ataggaaagc gacagatgtg attgtttcta ccgagaagag cattgtttcg
     5041 aataacagta tctcactgat aagctatctt attctttcga tcggttttcg tggcaatatc
     5101 gttcatatgc ggaaatatgt ggacactgaa ctgagtgaaa ttttaatacc tgataaaatt
     5161 ctgataaaaa tattgatgaa aaaaatcata agtttgtatc aatattgttt gtaatctaat
     5221 gactctaaat ttgctacaca agattgatca tacacagcca ttgagtttta agtcactcaa
     5281 attggctaac aggattgcaa ttatgaattg taaattatgc tcataaaatt tcaattctgc
     5341 tacaaaatgt tatgctactg aacatgactt taaaactgaa tgttaaaact agtggcaact
     5401 attagaaagg attgctattc acgctatatg gtgatatcga aatcatttcc attatttact
     5461 gagtaccatg aatcatctgt ttagctcact tgcacaaact gcccaaatgg gttcttgatc
     5521 attatctgag ttgatgatta cgggtttgtg agttatcgtt caattgaaaa cgatgatgat
     5581 aatgatattc tataagaggc aacgtatcga tcgatcgcat acatcgatca taattgtatt
     5641 tgaattgtag ttgagaagtg agcgaataac cgtctgtgag cgaattgatg ccaatatgct
     5701 tcaatactga ataaatgagg acaatcaatt taaccctctc ttttccatgg tggttccaga
     5761 gctccattcg ttttcgacaa acttgagaac cagatgaatg ctatgcaata catagttact
     5821 acaatatgat tctatactca gcagattaat ccaaaagtaa actaagagtt tcaatagtaa
     5881 aactattgat aaacaatcaa acaaaaaaat aaaccgattt cgaaaagtgt agaccagggc
     5941 tgcccaacgt acggcccgcg ggccgcatcc ggcccgcgac tccatttttt gcggcccgcg
     6001 gagggttcga agattcttct tatattttgc ccgctgaata ttctaccaac ccaaagttac
     6061 cataccaaga ctattaaatt ttcaatttag tttcaatatc atattctagc ttagtaatta
     6121 tttaaattgt tttgtttttt aaccttctta gatatcgaat tgtatggatg attcaaattc
     6181 aaaatctcga attcggagtg tttcaacaat atttctacca tgacttccat ctacagtgaa
     6241 acctccatga gtcgatattg aagggaccat cgactcatgg aaatatcgag tcatggaaca
     6301 gcaatccttt gcaaacctgc ttatagggat gatggtccct aaccatgtaa ttttgttttt
     6361 agtatgtata gtaatagtaa ccatgaaatt ttgtttttag tatggttcca tgagtcgata
     6421 tcgagtcatg gaacatcgac tcatggaggt atcactgtac attcattcat tgattcgaca
     6481 tttatccttg ataggattat agaaaaggcg ggcattttac aaaatcttga acaggatttt
     6541 ttaagaattc tggacacgac tttcgtggat aaaactgagt ttgctgtttc gctcgagaaa
     6601 aataaagcaa ttccttgagt ggatgggtaa aaaaaaaata attcagagag ccccatttga
     6661 catgacattt tatctcaact gagtactgcg attaaagaaa tgtgattttc tttaccactt
     6721 cctaccagga ttttcgtaaa agcgtgatca gagtttatca ttccggttca tgctaagaat
     6781 tttaagtcgg atactgacaa aattcggagt acgtttgaaa aaaattctag tatcattcta
     6841 ggatagattc tattctaaat attttggact atattatcat ataattttgt aagggatttt
     6901 cacaatgtcc agcagaatgc ttaattgctg ttaagctttt cttttttcga tttcttcaaa
     6961 tgaaattgac tccattcact gatttttgtc aggttttttg aacttcattt acaggctttt
     7021 gccagacttt atggaaatat ttgtgcgttt aaaaatgtgg cccgcgacac aaaattattt
     7081 ctgcgatttg gcccgcggtt caaaaaggtt gggcaggcct ggtgtagacc atttgcaaca
     7141 ttatttgctc aagcacttca agaaattgtc aatcaaagtc gtgggaaaaa gagggttaat
     7201 agctcttatc aaggaagatg ttttgtatgt gcgtggatcc aggatattca tttatcaaca
     7261 acgtgacttt tcccggagcg cacaaaacgt gcggccgcgt cgacatgccc gccgtgaccg
     7321 tcgagaaccc gctgacgctg ccccgcgtat ccgcacccgc cgacgccgtc gcacgtcccg
     7381 tgctcaccgt gaccaccgcg cccagcggtt tcgagggcga gggcttcccg gtgcgccgcg
     7441 cgttcgccgg gatcaactac cgccacctcg acccgttcat catgatggac cagatgggtg
     7501 aggtggagta cgcgcccggg gagcccaagg gcacgccctg gcacccgcac cgcggcttcg
     7561 agaccgtgac ctacatcgtc gacctaggac cccgggcgag ctcgaattaa ccattgtggg
     7621 aaccgtgcga tcaaacaaac gcgagatacc ggaagtactg aaaaacagtc gctccaggcc
     7681 agtgggaaca tcgatgtttt gttttgacgg accccttact ctcgtctcat ataaaccgaa
     7741 gccagctaag atggtatact tattatcatc ttgtgatgag gatgcttcta tcaacgaaag
     7801 taccggtaaa ccgcaaatgg ttatgtatta taatcaaact aaaggcggag tggacacgct
     7861 agaccaaatg tgttctgtga tgacctgcag taggaagacg aataggtggc ctatggcatt
     7921 attgtacgga atgataaaca ttgcctgcat aaattctttt attatataca gccataatgt
     7981 cagtagcaag ggagaaaagg ttcaaagtcg caaaaaattt atgagaaacc tttacatgag
     8041 cctgacgtca tcgtttatgc gtaagcgttt agaagctcct actttgaaga gatatttgcg
     8101 cgataatatc tctaatattt tgccaaatga agtgcctggt acatcagatg acagtactga
     8161 agagccagta atgaaaaaac gtacttactg tacttactgc ccctctaaaa taaggcgaaa
     8221 ggcaaatgca tcgtgcaaaa aatgcaaaaa agttatttgt cgagagcata atattgatat
     8281 gtgccaaagt tgtttctgac tgactaataa gtataatttg tttctattat gtataagtta
     8341 agctaattac ttattttata atacaacatg actgttttta aagtacaaaa taagtttatt
     8401 tttgtaaaag agagaatgtt taaaagtttt gttactttat agaagaaatt ttgagttttt
     8461 gttttttttt aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat
     8521 gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata
     8581 cagaccgata aaacacatgc gtcaatttta cgcatgatta tctttaacgt acgtcacaat
     8641 atgattatct ttctagggtt aaataatagt ttctaatttt tttattattc agcctgctgt
     8701 cgtgaatacc gtatatctca acgctgtctg tgagattgtc gtattctagc ctttttagtt
     8761 tttcgctcat cgacttgata ttgtccgaca cattttcgtc gatttgcgtt ttgatcaaag
     8821 acttgagcag agacacgtta atcaactgtt caaattgatc catattaacg atatcaaccc
     8881 gatgcgtata tggtgcgtaa aatatatttt ttaaccctct tatactttgc actctgcgtt
     8941 aatacgcgtt cgtgtacaga cgtaatcatg ttttcttttt tggataaaac tcctactgag
     9001 tttgacctca tattagaccc tcacaagttg caaaacgtgg cattttttac caatgaagaa
     9061 tttaaagtta ttttaaaaaa tttcatcaca gatttaaaga agaaccaaaa attaaattat
     9121 ttcaacagtt taatcgacca gttaatcaac gtgtacacag acgcgtcggc aaaaaacacg
     9181 cagcccgacg tgttggctaa aattattaaa tcaacttgtg ttatagtcac ggatttgccg
     9241 tccaacgtgt tcctcaaaaa gttgaagacc aacaagttta cggacactat taattatttg
     9301 attttgcccc acttcatttt gtgggatcac aattttgtta tatttttaaa caaagcttgg
     9361 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     9421 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     9481 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     9541 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     9601 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     9661 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     9721 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     9781 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     9841 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     9901 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     9961 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
    10021 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
    10081 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
    10141 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
    10201 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
    10261 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
    10321 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
    10381 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
    10441 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
    10501 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
    10561 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
    10621 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
    10681 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
    10741 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
    10801 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
    10861 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
    10921 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
    10981 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
    11041 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
    11101 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
    11161 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
    11221 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
    11281 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
    11341 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
    11401 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
    11461 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
    11521 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
    11581 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
    11641 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
    11701 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
    11761 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
    11821 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
    11881 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
    11941 tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat tcgagctcgg
    12001 tacccgggga tcctctagag tcgacgctcg cgcgacttgg tttgccattc tttagcgcgc
    12061 gtcgcgtcac acagcttggc cacaatgtgg tttttgtcaa acgaagattc tatgacgtgt
    12121 ttaaagttta ggtcgagtaa agcgcaaatc ttttttaacc ctagaaagat agtctgcgta
    12181 aaattgacgc atgcattctt gaaatattgc tctctctttc taaatagcgc gaatccgtcg
    12241 ctgtgcattt aggacatctc agtcgccgct tggagctccc gtgaggcgtg cttgtcaatg
    12301 cggtaagtgt cactgatttt gaactataac gaccgcgtga gtcaaaatga cgcatgatta
    12361 tcttttacgt gacttttaag atttaactca tacgataatt atattgttat ttcatgttct
    12421 acttacgtga taacttatta tatatatatt ttcttgttat agatatcgtg actaatatat
    12481 aataaaatgg gtagttcttt agacgatgag catatcctct ctgctcttct gcaaagcgat
    12541 gacgagcttg ttggtgagga ttctgacagt gaaatatcag atcacgtaag tgaagatgac
    12601 gtccagagcg atacagaaga agcgtttata gatgaggtac atgaagtgca gccaacgtca
    12661 agcggtagtg aaatattaga cgaacaaaat gttattgaac aaccaggttc ttcattggct
    12721 tctaacagaa tcttgacctt gccacagagg actattagag gtaagaataa acattgttgg
    12781 tcaacttcaa agtccacgag gcgtagccga gtctctgcac tgaacattgt cagatccgag
    12841 atcggccggc ctagaagctt taagatacat tgatgagttt ggacaaacca caactagaat
    12901 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
    12961 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
    13021 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
    13081 ttatgatcta gagtcctaca ggaacaggtg gtggcggccc tcggtgcgct cgtactgctc
    13141 cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc ttggagtcca cgtagtagta
    13201 gccgggcagc tgcacgggct tcttggccat gtagatggac ttgaactcca ccaggtagtg
    13261 gccgccgtcc ttcagcttca gggccttgtg gatctcgccc ttcagcacgc cgtcgcgggg
    13321 gtacaggcgc tcggtggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc
    13381 ggaggggaag ttcacgccga tgaacttcac cttgtagatg aagcagccgt cctgcaggga
    13441 ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt
    13501 gaagccctcg gggaaggaca gcttcttgta gtcggggatg tcggcggggt gcttcacgta
    13561 caccttggag ccgtactgga actgggggga caggatgtcc caggcgaagg gcagggggcc
    13621 gcccttggtc accttcagct tcacggtgtt gtggccctcg taggggcggc cctcgccctc
    13681 gccctcgatc tcgaactcgt ggccgttcac ggtgccctcc atgcgcacct tgaagcgcat
    13741 gaactccttg atgacgttct tggaggagcg caccatcacc agagacaggt tgcggcggcg
    13801 gttggatggc gtgggcgcgt tggcgttgtt ggaccggctc atgttgtgtc gctgtaacag
    13861 atgctgttca actgtgttta ccagatcgtt gcgggctgta tttataggcg cgataagcgg
    13921 gacgggcgcc tcgtgtccgg tcacgcgcat gagataacgc gcggctgata tggaggcgcg
    13981 tcctgttccg ataaggagtt gcgtccggct gcggttagca acacaggaag ctggcgtcct
    14041 gtcacgataa gacaacactc gtccggtccg ataatgtgat tcgtacgtga caggacgcga
    14101 cccgataagg ccggcctacg tgactgccga cacgtacttt tttgcactgc aaaaaggttc
    14161 aatgtgtggt agtgtatttg gagcgtatac aacggtgtag actatttatg taaaatagtc
    14221 tacgaaacgt agagtttgta ctatgtatgg gcccgcgtgc aaaagcgtgt ttttttgcag
    14281 tgcaaaaaag ttggtggtgg ggaggccacc gagtatgggc gcgccccggc cgttaactcg
    14341 aatcgctatc caagccagct ccattgtcgg catcgtgctc attctattga cgctggcaga
    14401 tttggttttg gcgctatggg acccgttcgg ttacaacaac atgtttccgc gcgagtttcc
    14461 cgacgacatg tcgcgcacgt tcctgactgc gtactttgag agtttcgaca acaccacgtc
    14521 cagagaaatc atagagttta tgcccgagtt cttttcggaa atggtcgaaa cggacgatga
    14581 cgccacgttt gaatctctat ttcatttatt agattatgtg gcatctttag aagttaattc
    14641 cgacggccaa atgttaaact tggaggaggg tgatgaaatt gaggattttg acgaatctac
    14701 tttggtgggg caagcgttag ccactagctc gctatacact cgcatggagt ttatgcagta
    14761 cacgtttagg caaaacacac tattgtctat gaacaaagaa aacaacaatt ttaatcaaat
    14821 aatcatgggt ttatttgcaa caaacacaat tgtggcgttt acagcatttg ttatacacac
    14881 agaactcata ttttttatat ttttcgtaat cttcctaatg atcacatttt attacataat
    14941 caaagaatcg tacgaatatt ataaaacaat tgatttgtta ttttaaaaac gattcattct
    15001 agttaattaa ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
    15061 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg
    15121 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
    15181 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca
    15241 ccttgatgcc gttcttctgc ttgtcggcca t
//
