LOCUS       sequence-193540-       14898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The promoter of AAEL010097 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 14898)
  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 14898)
  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: AAEL010097-Cas9
FEATURES             Location/Qualifiers
     source          1..14898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     348..365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    482..503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(518..523,524..541,542..548)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    556..572
                     /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     561..583
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     580..596
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     580..596
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   777..811
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    1479..1600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1490..1509)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1544..1563
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1714..2388,2389..2391,2392..2394))
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(1776..1799)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1974..1993)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2191..2211
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     CDS             complement(join(3629..4342,4343..4345,4346..4348))
                     /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(3674..3695)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4023..4042
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(4349..4402)
                     /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(4403..4450)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4451..8551)
                     /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(4451..8551)
                     /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(8576..8596)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8603..8668)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    11009..11078
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     repeat_region   12112..12174
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(12878..12895)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12878..12894)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12887..12909)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(13103..13122)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     13222..13244
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(13282..13300)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        13368..13472
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13473..13541,13542..14333)
                     /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(13691..13710)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(14504..14898,1..194)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
       61 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      301 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      361 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
      421 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
      481 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
      541 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga
      601 ttacgaattc gagctcggta cccggggatc ctctagagtc gacgctcgcg cgacttggtt
      661 tgccattctt tagcgcgcgt cgcgtcacac agcttggcca caatgtggtt tttgtcaaac
      721 gaagattcta tgacgtgttt aaagtttagg tcgagtaaag cgcaaatctt ttttaaccct
      781 agaaagatag tctgcgtaaa attgacgcat gcattcttga aatattgctc tctctttcta
      841 aatagcgcga atccgtcgct gtgcatttag gacatctcag tcgccgcttg gagctcccgt
      901 gaggcgtgct tgtcaatgcg gtaagtgtca ctgattttga actataacga ccgcgtgagt
      961 caaaatgacg catgattatc ttttacgtga cttttaagat ttaactcata cgataattat
     1021 attgttattt catgttctac ttacgtgata acttattata tatatatttt cttgttatag
     1081 atatcgtgac taatatataa taaaatgggt agttctttag acgatgagca tatcctctct
     1141 gctcttctgc aaagcgatga cgagcttgtt ggtgaggatt ctgacagtga aatatcagat
     1201 cacgtaagtg aagatgacgt ccagagcgat acagaagaag cgtttataga tgaggtacat
     1261 gaagtgcagc caacgtcaag cggtagtgaa atattagacg aacaaaatgt tattgaacaa
     1321 ccaggttctt cattggcttc taacagaatc ttgaccttgc cacagaggac tattagaggt
     1381 aagaataaac attgttggtc aacttcaaag tccacgaggc gtagccgagt ctctgcactg
     1441 aacattgtca gatccgagat cggccggcct agaagcttta agatacattg atgagtttgg
     1501 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1561 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1621 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1681 caaatgtggt atggctgatt atgatctaga gtcctacagg aacaggtggt ggcggccctc
     1741 ggtgcgctcg tactgctcca cgatggtgta gtcctcgttg tgggaggtga tgtccagctt
     1801 ggagtccacg tagtagtagc cgggcagctg cacgggcttc ttggccatgt agatggactt
     1861 gaactccacc aggtagtggc cgccgtcctt cagcttcagg gccttgtgga tctcgccctt
     1921 cagcacgccg tcgcgggggt acaggcgctc ggtggaggcc tcccagccca tggtcttctt
     1981 ctgcattacg gggccgtcgg aggggaagtt cacgccgatg aacttcacct tgtagatgaa
     2041 gcagccgtcc tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt
     2101 catcacgcgc tcccacttga agccctcggg gaaggacagc ttcttgtagt cggggatgtc
     2161 ggcggggtgc ttcacgtaca ccttggagcc gtactggaac tggggggaca ggatgtccca
     2221 ggcgaagggc agggggccgc ccttggtcac cttcagcttc acggtgttgt ggccctcgta
     2281 ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg tgccctccat
     2341 gcgcaccttg aagcgcatga actccttgat gacgttcttg gaggagcgca ccatcaccag
     2401 agacaggttg cggcggcggt tggatggcgt gggcgcgttg gcgttgttgg accggctcat
     2461 gttgtgtcgc tgtaacagat gctgttcaac tgtgtttacc agatcgttgc gggctgtatt
     2521 tataggcgcg ataagcggga cgggcgcctc gtgtccggtc acgcgcatga gataacgcgc
     2581 ggctgatatg gaggcgcgtc ctgttccgat aaggagttgc gtccggctgc ggttagcaac
     2641 acaggaagct ggcgtcctgt cacgataaga caacactcgt ccggtccgat aatgtgattc
     2701 gtacgtgaca ggacgcgacc cgataaggcc ggcctacgtg actgccgaca cgtacttttt
     2761 tgcactgcaa aaaggttcaa tgtgtggtag tgtatttgga gcgtatacaa cggtgtagac
     2821 tatttatgta aaatagtcta cgaaacgtag agtttgtact atgtatgggc ccgcgtgcaa
     2881 aagcgtgttt ttttgcagtg caaaaaagtt ggtggtgggg aggccaccga gtatgggcgc
     2941 gccccggccg ttaactcgaa tcgctatcca agccagctcc attgtcggca tcgtgctcat
     3001 tctattgacg ctggcagatt tggttttggc gctatgggac ccgttcggtt acaacaacat
     3061 gtttccgcgc gagtttcccg acgacatgtc gcgcacgttc ctgactgcgt actttgagag
     3121 tttcgacaac accacgtcca gagaaatcat agagtttatg cccgagttct tttcggaaat
     3181 ggtcgaaacg gacgatgacg ccacgtttga atctctattt catttattag attatgtggc
     3241 atctttagaa gttaattccg acggccaaat gttaaacttg gaggagggtg atgaaattga
     3301 ggattttgac gaatctactt tggtggggca agcgttagcc actagctcgc tatacactcg
     3361 catggagttt atgcagtaca cgtttaggca aaacacacta ttgtctatga acaaagaaaa
     3421 caacaatttt aatcaaataa tcatgggttt atttgcaaca aacacaattg tggcgtttac
     3481 agcatttgtt atacacacag aactcatatt ttttatattt ttcgtaatct tcctaatgat
     3541 cacattttat tacataatca aagaatcgta cgaatattat aaaacaattg atttgttatt
     3601 ttaaaaacga ttcattctag ttaattaatt acttgtacag ctcgtccatg ccgagagtga
     3661 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     3721 tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     3781 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     3841 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     3901 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     3961 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     4021 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     4081 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     4141 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgctggtgg
     4201 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     4261 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     4321 tgaacagctc ctcgcccttg ctcaccatag ggccgggatt ctcctccacg tcaccgcatg
     4381 ttagaagact tcctctgccc tcctttttct tttttgcctg gccggccttt ttcgtggccg
     4441 ccggcctttt gtcgcctccc agctgagaca ggtcgatccg tgtctcgtac aggccggtga
     4501 tgctctggtg gatcagggtg gcgtccagca cctctttggt gctggtgtac ctcttccggt
     4561 cgatggtggt gtcaaagtac ttgaaggcgg caggggctcc cagattggtc agggtaaaca
     4621 ggtggatgat attctcggcc tgctctctga tgggcttatc ccggtgcttg ttgtaggcgg
     4681 acagcacttt gtccagatta gcgtcggcca ggatcactct cttggagaac tcgctgatct
     4741 gctcgatgat ctcgtccagg tagtgcttgt gctgttccac aaacagctgt ttctgctcat
     4801 tatcctcggg ggagcccttc agcttctcat agtggctggc caggtacagg aagttcacat
     4861 atttggaggg cagggccagt tcgtttccct tctgcagttc gccggcagag gccagcattc
     4921 tcttccggcc gttttccagc tcgaacaggg agtacttagg cagcttgatg atcaggtcct
     4981 ttttcacttc tttgtagccc ttggcttcca gaaagtcgat gggattcttc tcgaagctgc
     5041 ttctttccat gatggtgatc cccagcagct ctttcacact cttcagtttc ttggacttgc
     5101 ccttttccac tttggccacc accagcacag aataggccac ggtggggctg tcgaagccgc
     5161 cgtacttctt agggtcccag tccttctttc tggcgatcag cttatcgctg ttcctcttgg
     5221 gcaggataga ctctttgctg aagccgcctg tctgcacctc ggtctttttc acgatattca
     5281 cttggggcat gctcagcact ttccgcacgg tggcaaaatc ccggccctta tcccacacga
     5341 tctccccggt ttcgccgttt gtctcgatca gaggccgctt ccggatctcg ccgttggcca
     5401 gggtaatctc ggtcttgaaa aagttcatga tgttgctgta gaagaagtac ttggcggtag
     5461 ccttgccgat ttcctgctcg ctcttggcga tcatcttccg cacgtcgtac accttgtagt
     5521 cgccgtacac gaactcgctt tccagcttag ggtacttttt gatcagggcg gttcccacga
     5581 cggcgttcag gtaggcgtcg tgggcgtggt ggtagttgtt gatctcgcgc actttgtaaa
     5641 actggaaatc cttccggaaa tcggacacca gcttggactt cagggtgatc actttcactt
     5701 cccggatcag cttgtcattc tcgtcgtact tagtgttcat ccgggagtcc aggatctgtg
     5761 ccacgtgctt tgtgatctgc cgggtttcca ccagctgtct cttgatgaag ccggccttat
     5821 ccagttcgct caggccgcct ctctcggcct tggtcagatt gtcgaacttt ctctgggtaa
     5881 tcagcttggc gttcagcagc tgccgccagt agttcttcat cttcttcacg acctcttcgg
     5941 agggcacgtt gtcgctcttg ccccggttct tgtcgcttct ggtcagcacc ttgttgtcga
     6001 tggagtcgtc cttcagaaag ctctgaggca cgatatggtc cacatcgtag tcggacagcc
     6061 ggttgatgtc cagttcctgg tccacgtaca tatcccgccc attctgcagg tagtacaggt
     6121 acagcttctc gttctgcagc tgggtgtttt ccacggggtg ttctttcagg atctggctgc
     6181 ccagctcttt gatgccctct tcgatccgct tcattctctc gcggctgttc ttctgtccct
     6241 tctgggtggt ctggttctct ctggccattt cgatcacgat gttctcgggc ttgtgccggc
     6301 ccatcacttt cacgagctcg tccaccacct tcactgtctg caggatgccc ttcttaatgg
     6361 cggggctgcc ggccagattg gcaatgtgct cgtgcaggct atcgccctgg ccggacacct
     6421 gggctttctg gatgtcctct ttaaaggtca ggctgtcgtc gtggatcagc tgcatgaagt
     6481 ttctgttggc gaagccgtcg gacttcagga aatccaggat tgtcttgccg gactgcttgt
     6541 cccggatgcc gttgatcagc ttccggctca gcctgcccca gccggtgtat ctccgccgct
     6601 tcagctgctt catcactttg tcgtcgaaca ggtgggcata ggttttcagc cgttcctcga
     6661 tcatctctct gtcctcaaac agtgtcaggg tcagcacgat atcttccaga atgtcctcgt
     6721 tttcctcatt gtccaggaag tccttgtcct tgataatttt cagcagatcg tggtatgtgc
     6781 ccagggaggc gttgaaccga tcttccacgc cggagatttc cacggagtcg aagcactcga
     6841 ttttcttgaa gtagtcctct ttcagctgct tcacggtcac tttccggttg gtcttgaaca
     6901 gcaggtccac gatggccttt ttctgctcgc cgctcaggaa ggcgggcttt ctcattccct
     6961 cggtcacgta tttcactttg gtcagctcgt tatacacggt gaagtactcg tacagcaggc
     7021 tgtgcttggg cagcaccttc tcgttgggca ggttcttatc gaagttggtc atccgctcga
     7081 tgaagctctg ggcggaagcg cccttgtcca ccacttcctc gaagttccag ggggtgatgg
     7141 tttcctcgct ctttctggtc atccaggcga atctgctgtt tcccctggcc agagggccca
     7201 cgtagtaggg gatgcggaag gtcaggatct tctcgatctt ttcccggttg tccttcagga
     7261 atgggtaaaa atcttcctgc cgccgcagaa tggcgtgcag ctctcccagg tggatctggt
     7321 gggggatgct gccgttgtcg aaggtccgct gcttccgcag caggtcctct ctgttcagct
     7381 tcacgagcag ttcctcggtg ccgtccatct tttccaggat gggcttgatg aacttgtaga
     7441 actcttcctg gctggctccg ccgtcaatgt agccggcgta gccgttcttg ctctggtcga
     7501 agaaaatctc tttgtacttc tcaggcagct gctgccgcac gagagctttc agcagggtca
     7561 ggtcctggtg gtgctcgtcg tatctcttga tcatagaggc gctcaggggg gccttggtga
     7621 tctcggtgtt cactctcagg atgtcgctca gcaggatggc gtcggacagg ttcttggcgg
     7681 ccagaaacag gtcggcgtac tggtcgccga tctgggccag caggttgtcc aggtcgtcgt
     7741 cgtaggtgtc cttgctcagc tgcagtttgg catcctcggc caggtcgaag ttgctcttga
     7801 agttgggggt caggcccagg ctcagggcaa tcaggtttcc gaacaggcca ttcttcttct
     7861 cgccgggcag ctgggcgatc agattttcca gccgtctgct cttgctcagt ctggcagaca
     7921 ggatggcctt ggcgtccacg ccgctggcgt tgatggggtt ttcctcgaac agctggttgt
     7981 aggtctgcac cagctggatg aacagcttgt ccacgtcgct gttgtcgggg ttcaggtcgc
     8041 cctcgatcag gaagtggccc cggaacttga tcatgtgggc cagggccaga tagatcagcc
     8101 gcaggtcggc cttgtcggtg ctgtccacca gtttctttct caggtggtag atggtggggt
     8161 acttctcgtg gtaggccacc tcgtccacga tgttgccgaa gatggggtgc cgctcgtgct
     8221 tcttatcctc ttccaccagg aaggactctt ccagtctgtg gaagaagctg tcgtccacct
     8281 tggccatctc gttgctgaag atctcttgca gatagcagat ccggttcttc cgtctggtgt
     8341 atcttcttct ggcggttctc ttcagccggg tggcctcggc tgtttcgccg ctgtcgaaca
     8401 gcagggctcc gatcaggttc ttcttgatgc tgtgccggtc ggtgttgccc agcaccttga
     8461 atttcttgct gggcaccttg tactcgtcgg tgatcacggc ccagcccaca gagttggtgc
     8521 cgatgtccag gccgatgctg tacttcttgt cggctgctgg gactccgtgg ataccgacct
     8581 tccgcttctt ctttggggcc atcttatcgt catcgtcttt gtaatcaata tcatgatcct
     8641 tgtagtctcc gtcgtggtcc ttatagtcca tctcgagttt cactctgtag acaaaagata
     8701 acctctttta tcccagcaac ctgcttttcc tccgaaaagg aaaattccac aaaatggtca
     8761 catggtggaa aaaagaacga cacttaccaa tactctttcg aagaaattcg caattttcac
     8821 acaggtttta atcactgttt tcacgcacgc ttttagcgac ccgtttgtgt tttagatttt
     8881 cacttttgag gatattccac agaacacttt tctgaaactt tccttatttt tcttccaacg
     8941 cggcgatgct cgtccttggc ttgatgaact tgaagtgaag gttggcaagg tcgatctggt
     9001 ctacagcaaa aaaaaaattc ggacgaaacg ggagcaaaat tccgttagtg tcggtgcggt
     9061 actagaatga gttacagttg attacgtgcg ggcagtgccg ggttttgtgg aatatccggc
     9121 ggagaaattc gaatattaga ttaaatgtga ccattgatgg aagtgaaaat ttatactcga
     9181 tgcggtatta ttggaatatt tgagatgcca atggaatata ttgcaagaaa tgcgaaaaat
     9241 gtcgaccgtt aatatttttt taatcatgca ctcaccacta ccagacaggg atgccaggta
     9301 atttattttt atgtattgat ttcaatgcac ggaaaagtgt catccaatgt tttccacctt
     9361 aaatttttca aaaataacac tgcggacctt tttttcgcct caagcatcaa aatatctata
     9421 tttacttaag gctaagtagc ccgtcattcg ttttggcaac aatgatgacc tttcagcttg
     9481 catttcaaag tgataaaact cagtcttgat agtttatatt gacttgaaaa agtatctctt
     9541 tacgcactaa gatgcataaa gtatgctgta tgctgatact ttttcagctg tgtcagtgca
     9601 aaaccaactg attttctttg attcgaaatc gtgagatgag ttagggccca ttcacaaatt
     9661 tcataacgct agagggggtg ggtagcgatg ttacggccca tacaaaaatt gaataacatt
     9721 catacaaaaa gcgttacaag ggggtgggtg ggtgcccaaa atacgcttcc aatcgcaaaa
     9781 cgcaacatac tgttagccat ctatactttg gtagcgcaac tagtcagatg atgagagatt
     9841 tgatttttca cgcatccatc acatgccgct gtggggaaaa atgttttatt ttcctgatat
     9901 aagacggttc acagtgaaac agttaggcgg aaaaggtagg aatgtccatt ctctatgttc
     9961 tacatacgtt tgtttttttg tattattgca ctacttttat aaaaattgag atccggctgg
    10021 taggagcact agtttgaggt tggaaaacca ggatatgtga ggttagattc attggaactt
    10081 tcgcctgagc gatttttgcg cttgaataaa acacgcttcg gaacattctc cgctgcccct
    10141 aaaaatacca ttgggtagat aaatgaattc tctaccattg gatctcattc aatttcatac
    10201 ataagtgaag taagcggtga aaataaatat tttgttccga atgttttgaa accatgaatg
    10261 agtttgacat tgctctcgtc aaccatcatc atgacgacag cagcacaccc caccggacga
    10321 attgtcgtat tgggagcgaa tggccaattt cagcgttatg aaataaatga atgaaccctt
    10381 agcaacaatc atcaacgacg cgtacaaatt tcaatgacgg cctacttcgc cttaatttag
    10441 ggttgtcgtt ttataaattg tataagccat tttacgagac gtttcggaac agctgatcgc
    10501 cgaaacggcg tcaattgaca ggaggcctgg gattaaatcc agcgtgattt ttaacaacgc
    10561 ctcatcttac caaacgagga catgaagaaa atgacaaaaa agagatccaa aaatgtttgt
    10621 tactgcaact taacacctag ttattgtatc agctacctct ttttcaccta tatttcacat
    10681 aaaaaggcat ttttgtgatc agaaatattt taatattttt tctccattta agttttcgcc
    10741 tgtataaaaa gctacgcttg aaatgcgcac agtcatcaac catcatcatc gccataactg
    10801 acgacggcgg ccgcgtcgac atgcccgccg tgaccgtcga gaacccgctg acgctgcccc
    10861 gcgtatccgc acccgccgac gccgtcgcac gtcccgtgct caccgtgacc accgcgccca
    10921 gcggtttcga gggcgagggc ttcccggtgc gccgcgcgtt cgccgggatc aactaccgcc
    10981 acctcgaccc gttcatcatg atggaccaga tgggtgaggt ggagtacgcg cccggggagc
    11041 ccaagggcac gccctggcac ccgcaccgcg gcttcgagac cgtgacctac atcgtcgacc
    11101 taggaccccg ggcgagctcg aattaaccat tgtgggaacc gtgcgatcaa acaaacgcga
    11161 gataccggaa gtactgaaaa acagtcgctc caggccagtg ggaacatcga tgttttgttt
    11221 tgacggaccc cttactctcg tctcatataa accgaagcca gctaagatgg tatacttatt
    11281 atcatcttgt gatgaggatg cttctatcaa cgaaagtacc ggtaaaccgc aaatggttat
    11341 gtattataat caaactaaag gcggagtgga cacgctagac caaatgtgtt ctgtgatgac
    11401 ctgcagtagg aagacgaata ggtggcctat ggcattattg tacggaatga taaacattgc
    11461 ctgcataaat tcttttatta tatacagcca taatgtcagt agcaagggag aaaaggttca
    11521 aagtcgcaaa aaatttatga gaaaccttta catgagcctg acgtcatcgt ttatgcgtaa
    11581 gcgtttagaa gctcctactt tgaagagata tttgcgcgat aatatctcta atattttgcc
    11641 aaatgaagtg cctggtacat cagatgacag tactgaagag ccagtaatga aaaaacgtac
    11701 ttactgtact tactgcccct ctaaaataag gcgaaaggca aatgcatcgt gcaaaaaatg
    11761 caaaaaagtt atttgtcgag agcataatat tgatatgtgc caaagttgtt tctgactgac
    11821 taataagtat aatttgtttc tattatgtat aagttaagct aattacttat tttataatac
    11881 aacatgactg tttttaaagt acaaaataag tttatttttg taaaagagag aatgtttaaa
    11941 agttttgtta ctttatagaa gaaattttga gtttttgttt ttttttaata aataaataaa
    12001 cataaataaa ttgtttgttg aatttattat tagtatgtaa gtgtaaatat aataaaactt
    12061 aatatctatt caaattaata aataaacctc gatatacaga ccgataaaac acatgcgtca
    12121 attttacgca tgattatctt taacgtacgt cacaatatga ttatctttct agggttaaat
    12181 aatagtttct aattttttta ttattcagcc tgctgtcgtg aataccgtat atctcaacgc
    12241 tgtctgtgag attgtcgtat tctagccttt ttagtttttc gctcatcgac ttgatattgt
    12301 ccgacacatt ttcgtcgatt tgcgttttga tcaaagactt gagcagagac acgttaatca
    12361 actgttcaaa ttgatccata ttaacgatat caacccgatg cgtatatggt gcgtaaaata
    12421 tattttttaa ccctcttata ctttgcactc tgcgttaata cgcgttcgtg tacagacgta
    12481 atcatgtttt cttttttgga taaaactcct actgagtttg acctcatatt agaccctcac
    12541 aagttgcaaa acgtggcatt ttttaccaat gaagaattta aagttatttt aaaaaatttc
    12601 atcacagatt taaagaagaa ccaaaaatta aattatttca acagtttaat cgaccagtta
    12661 atcaacgtgt acacagacgc gtcggcaaaa aacacgcagc ccgacgtgtt ggctaaaatt
    12721 attaaatcaa cttgtgttat agtcacggat ttgccgtcca acgtgttcct caaaaagttg
    12781 aagaccaaca agtttacgga cactattaat tatttgattt tgccccactt cattttgtgg
    12841 gatcacaatt ttgttatatt tttaaacaaa gcttggcact ggccgtcgtt ttacaacgtc
    12901 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
    12961 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
    13021 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
    13081 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
    13141 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
    13201 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
    13261 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
    13321 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
    13381 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
    13441 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
    13501 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
    13561 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
    13621 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
    13681 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
    13741 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
    13801 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
    13861 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
    13921 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
    13981 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
    14041 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
    14101 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
    14161 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
    14221 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
    14281 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
    14341 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
    14401 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
    14461 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
    14521 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
    14581 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
    14641 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
    14701 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
    14761 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
    14821 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
    14881 acctacagcg tgagctat
//
