LOCUS       sequence-194325-       14155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The promoter of AAEL003877 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 14155)
  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 14155)
  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: AAEL003877-Cas9
FEATURES             Location/Qualifiers
     source          1..14155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..197
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(504..557)
                     /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(558..605)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(606..4706)
                     /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(606..4706)
                     /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(4731..4751)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4758..4823)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    6421..6490
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     repeat_region   7524..7586
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8290..8307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8290..8306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8299..8321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(8515..8534)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8634..8656
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8694..8712)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8780..8884
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8885..8953,8954..9745)
                     /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(9103..9122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9916..10504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10405..10424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10658..10675
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10792..10813
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10828..10833,10834..10851,10852..10858)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10866..10882
                     /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     10871..10893
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10890..10906
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10890..10906
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   11087..11121
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    11789..11910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11800..11819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     11854..11873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(12024..12698,12699..12701,12702..12704))
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(12086..12109)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(12284..12303)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     12501..12521
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     CDS             complement(join(13939..14155,1..497,498..500,501..503))
                     /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(13984..14005)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
       61 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      121 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      181 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
      241 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
      301 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgct ggtggtgcag
      361 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
      421 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
      481 agctcctcgc ccttgctcac catagggccg ggattctcct ccacgtcacc gcatgttaga
      541 agacttcctc tgccctcctt tttctttttt gcctggccgg cctttttcgt ggccgccggc
      601 cttttgtcgc ctcccagctg agacaggtcg atccgtgtct cgtacaggcc ggtgatgctc
      661 tggtggatca gggtggcgtc cagcacctct ttggtgctgg tgtacctctt ccggtcgatg
      721 gtggtgtcaa agtacttgaa ggcggcaggg gctcccagat tggtcagggt aaacaggtgg
      781 atgatattct cggcctgctc tctgatgggc ttatcccggt gcttgttgta ggcggacagc
      841 actttgtcca gattagcgtc ggccaggatc actctcttgg agaactcgct gatctgctcg
      901 atgatctcgt ccaggtagtg cttgtgctgt tccacaaaca gctgtttctg ctcattatcc
      961 tcgggggagc ccttcagctt ctcatagtgg ctggccaggt acaggaagtt cacatatttg
     1021 gagggcaggg ccagttcgtt tcccttctgc agttcgccgg cagaggccag cattctcttc
     1081 cggccgtttt ccagctcgaa cagggagtac ttaggcagct tgatgatcag gtcctttttc
     1141 acttctttgt agcccttggc ttccagaaag tcgatgggat tcttctcgaa gctgcttctt
     1201 tccatgatgg tgatccccag cagctctttc acactcttca gtttcttgga cttgcccttt
     1261 tccactttgg ccaccaccag cacagaatag gccacggtgg ggctgtcgaa gccgccgtac
     1321 ttcttagggt cccagtcctt ctttctggcg atcagcttat cgctgttcct cttgggcagg
     1381 atagactctt tgctgaagcc gcctgtctgc acctcggtct ttttcacgat attcacttgg
     1441 ggcatgctca gcactttccg cacggtggca aaatcccggc ccttatccca cacgatctcc
     1501 ccggtttcgc cgtttgtctc gatcagaggc cgcttccgga tctcgccgtt ggccagggta
     1561 atctcggtct tgaaaaagtt catgatgttg ctgtagaaga agtacttggc ggtagccttg
     1621 ccgatttcct gctcgctctt ggcgatcatc ttccgcacgt cgtacacctt gtagtcgccg
     1681 tacacgaact cgctttccag cttagggtac tttttgatca gggcggttcc cacgacggcg
     1741 ttcaggtagg cgtcgtgggc gtggtggtag ttgttgatct cgcgcacttt gtaaaactgg
     1801 aaatccttcc ggaaatcgga caccagcttg gacttcaggg tgatcacttt cacttcccgg
     1861 atcagcttgt cattctcgtc gtacttagtg ttcatccggg agtccaggat ctgtgccacg
     1921 tgctttgtga tctgccgggt ttccaccagc tgtctcttga tgaagccggc cttatccagt
     1981 tcgctcaggc cgcctctctc ggccttggtc agattgtcga actttctctg ggtaatcagc
     2041 ttggcgttca gcagctgccg ccagtagttc ttcatcttct tcacgacctc ttcggagggc
     2101 acgttgtcgc tcttgccccg gttcttgtcg cttctggtca gcaccttgtt gtcgatggag
     2161 tcgtccttca gaaagctctg aggcacgata tggtccacat cgtagtcgga cagccggttg
     2221 atgtccagtt cctggtccac gtacatatcc cgcccattct gcaggtagta caggtacagc
     2281 ttctcgttct gcagctgggt gttttccacg gggtgttctt tcaggatctg gctgcccagc
     2341 tctttgatgc cctcttcgat ccgcttcatt ctctcgcggc tgttcttctg tcccttctgg
     2401 gtggtctggt tctctctggc catttcgatc acgatgttct cgggcttgtg ccggcccatc
     2461 actttcacga gctcgtccac caccttcact gtctgcagga tgcccttctt aatggcgggg
     2521 ctgccggcca gattggcaat gtgctcgtgc aggctatcgc cctggccgga cacctgggct
     2581 ttctggatgt cctctttaaa ggtcaggctg tcgtcgtgga tcagctgcat gaagtttctg
     2641 ttggcgaagc cgtcggactt caggaaatcc aggattgtct tgccggactg cttgtcccgg
     2701 atgccgttga tcagcttccg gctcagcctg ccccagccgg tgtatctccg ccgcttcagc
     2761 tgcttcatca ctttgtcgtc gaacaggtgg gcataggttt tcagccgttc ctcgatcatc
     2821 tctctgtcct caaacagtgt cagggtcagc acgatatctt ccagaatgtc ctcgttttcc
     2881 tcattgtcca ggaagtcctt gtccttgata attttcagca gatcgtggta tgtgcccagg
     2941 gaggcgttga accgatcttc cacgccggag atttccacgg agtcgaagca ctcgattttc
     3001 ttgaagtagt cctctttcag ctgcttcacg gtcactttcc ggttggtctt gaacagcagg
     3061 tccacgatgg cctttttctg ctcgccgctc aggaaggcgg gctttctcat tccctcggtc
     3121 acgtatttca ctttggtcag ctcgttatac acggtgaagt actcgtacag caggctgtgc
     3181 ttgggcagca ccttctcgtt gggcaggttc ttatcgaagt tggtcatccg ctcgatgaag
     3241 ctctgggcgg aagcgccctt gtccaccact tcctcgaagt tccagggggt gatggtttcc
     3301 tcgctctttc tggtcatcca ggcgaatctg ctgtttcccc tggccagagg gcccacgtag
     3361 taggggatgc ggaaggtcag gatcttctcg atcttttccc ggttgtcctt caggaatggg
     3421 taaaaatctt cctgccgccg cagaatggcg tgcagctctc ccaggtggat ctggtggggg
     3481 atgctgccgt tgtcgaaggt ccgctgcttc cgcagcaggt cctctctgtt cagcttcacg
     3541 agcagttcct cggtgccgtc catcttttcc aggatgggct tgatgaactt gtagaactct
     3601 tcctggctgg ctccgccgtc aatgtagccg gcgtagccgt tcttgctctg gtcgaagaaa
     3661 atctctttgt acttctcagg cagctgctgc cgcacgagag ctttcagcag ggtcaggtcc
     3721 tggtggtgct cgtcgtatct cttgatcata gaggcgctca ggggggcctt ggtgatctcg
     3781 gtgttcactc tcaggatgtc gctcagcagg atggcgtcgg acaggttctt ggcggccaga
     3841 aacaggtcgg cgtactggtc gccgatctgg gccagcaggt tgtccaggtc gtcgtcgtag
     3901 gtgtccttgc tcagctgcag tttggcatcc tcggccaggt cgaagttgct cttgaagttg
     3961 ggggtcaggc ccaggctcag ggcaatcagg tttccgaaca ggccattctt cttctcgccg
     4021 ggcagctggg cgatcagatt ttccagccgt ctgctcttgc tcagtctggc agacaggatg
     4081 gccttggcgt ccacgccgct ggcgttgatg gggttttcct cgaacagctg gttgtaggtc
     4141 tgcaccagct ggatgaacag cttgtccacg tcgctgttgt cggggttcag gtcgccctcg
     4201 atcaggaagt ggccccggaa cttgatcatg tgggccaggg ccagatagat cagccgcagg
     4261 tcggccttgt cggtgctgtc caccagtttc tttctcaggt ggtagatggt ggggtacttc
     4321 tcgtggtagg ccacctcgtc cacgatgttg ccgaagatgg ggtgccgctc gtgcttctta
     4381 tcctcttcca ccaggaagga ctcttccagt ctgtggaaga agctgtcgtc caccttggcc
     4441 atctcgttgc tgaagatctc ttgcagatag cagatccggt tcttccgtct ggtgtatctt
     4501 cttctggcgg ttctcttcag ccgggtggcc tcggctgttt cgccgctgtc gaacagcagg
     4561 gctccgatca ggttcttctt gatgctgtgc cggtcggtgt tgcccagcac cttgaatttc
     4621 ttgctgggca ccttgtactc gtcggtgatc acggcccagc ccacagagtt ggtgccgatg
     4681 tccaggccga tgctgtactt cttgtcggct gctgggactc cgtggatacc gaccttccgc
     4741 ttcttctttg gggccatctt atcgtcatcg tctttgtaat caatatcatg atccttgtag
     4801 tctccgtcgt ggtccttata gtccatctcg agattcgttg aaatctctgt tgagcagaaa
     4861 aagaaacgag gaaacgcttc agtaattggt tgtgaaatgc aaactctcat ttgatattga
     4921 ttcattgcct ttggcttcga gcacgacacg acaggtttta aacttgtttt gctttgtctg
     4981 cgtttgcagt cgcaggccaa gtgaaaaata tacacttgaa ggtgatgacg tcacaacaac
     5041 gccctacttt ttagtgaaaa attaacttgt tttcgacttt gaactacgta gtttggaaat
     5101 tgcgtatctt caagttttca cgatttcctc aaggtttttc tcgatatgtg ttaatattac
     5161 cttaatgggt aattatcacc acaatattta attttagaaa tatggcgaat tcgcgcttgc
     5221 tctagaaatt taattcaatt agcagcgatg attcaacgaa atatgattat cgccgtgaat
     5281 cacaattggg ttttatcaat gatgatgaaa ctgcgttgca aattttcact aatcactcaa
     5341 agctcaatag tcgccatctt ggaaaatagt ttgctcattc aaagacaaag gaatcattca
     5401 ccaaactagt tttcgctcat agctataatt tcatcattta atttacctac cttcactaga
     5461 agattccctt caccgaaatg cactttcaca aatttaatag taattgtcct ttgaacaaag
     5521 ctttgttcac tctgaaattt tctcctctgg ctaattggat cactcttttt cactagagac
     5581 ttcacttcac ttgcactggc actgcttact tggaccgcgt aatgttcact ccactaggaa
     5641 acgtattcga ttgagctggt tttgcctttg caggggcgtt ttatagacac tgccgtagtg
     5701 gtggttgtac ttctagaaaa tttctgcaat ccattcatac atatacctct gttcggttgg
     5761 atggctctaa atcgatcgac tatgggtacc atccctgtgt ctgaatggac agcaaaacgt
     5821 gcttgtgtct gttagtcgtt cattctgtac cttgaacgat gcagttcaac ttctggcaaa
     5881 gacgtcaatg tacctaccat tcgtgtatat ggcatacaga gaaagatgtg cgaatgcctt
     5941 tttcgataga gaaaggattt cctgatttga tcgacaattt cggtagctat tcttagcttc
     6001 gaatgtaatc tgataaccaa atccagagac aaagcagtaa tttgggaatt tcacttgaat
     6061 cattctaatt gtatcatttg ctgtattagt gcagtcagca aagtgacgtc aacccttcta
     6121 aatcgatata cttctgggaa gctttctttc ttgtctggct cagctggtgc caaggcaaat
     6181 tataattgga ttcaatgcac aagctacatg taaagatagc ggccgcgtcg acatgcccgc
     6241 cgtgaccgtc gagaacccgc tgacgctgcc ccgcgtatcc gcacccgccg acgccgtcgc
     6301 acgtcccgtg ctcaccgtga ccaccgcgcc cagcggtttc gagggcgagg gcttcccggt
     6361 gcgccgcgcg ttcgccggga tcaactaccg ccacctcgac ccgttcatca tgatggacca
     6421 gatgggtgag gtggagtacg cgcccgggga gcccaagggc acgccctggc acccgcaccg
     6481 cggcttcgag accgtgacct acatcgtcga cctaggaccc cgggcgagct cgaattaacc
     6541 attgtgggaa ccgtgcgatc aaacaaacgc gagataccgg aagtactgaa aaacagtcgc
     6601 tccaggccag tgggaacatc gatgttttgt tttgacggac cccttactct cgtctcatat
     6661 aaaccgaagc cagctaagat ggtatactta ttatcatctt gtgatgagga tgcttctatc
     6721 aacgaaagta ccggtaaacc gcaaatggtt atgtattata atcaaactaa aggcggagtg
     6781 gacacgctag accaaatgtg ttctgtgatg acctgcagta ggaagacgaa taggtggcct
     6841 atggcattat tgtacggaat gataaacatt gcctgcataa attcttttat tatatacagc
     6901 cataatgtca gtagcaaggg agaaaaggtt caaagtcgca aaaaatttat gagaaacctt
     6961 tacatgagcc tgacgtcatc gtttatgcgt aagcgtttag aagctcctac tttgaagaga
     7021 tatttgcgcg ataatatctc taatattttg ccaaatgaag tgcctggtac atcagatgac
     7081 agtactgaag agccagtaat gaaaaaacgt acttactgta cttactgccc ctctaaaata
     7141 aggcgaaagg caaatgcatc gtgcaaaaaa tgcaaaaaag ttatttgtcg agagcataat
     7201 attgatatgt gccaaagttg tttctgactg actaataagt ataatttgtt tctattatgt
     7261 ataagttaag ctaattactt attttataat acaacatgac tgtttttaaa gtacaaaata
     7321 agtttatttt tgtaaaagag agaatgttta aaagttttgt tactttatag aagaaatttt
     7381 gagtttttgt ttttttttaa taaataaata aacataaata aattgtttgt tgaatttatt
     7441 attagtatgt aagtgtaaat ataataaaac ttaatatcta ttcaaattaa taaataaacc
     7501 tcgatataca gaccgataaa acacatgcgt caattttacg catgattatc tttaacgtac
     7561 gtcacaatat gattatcttt ctagggttaa ataatagttt ctaatttttt tattattcag
     7621 cctgctgtcg tgaataccgt atatctcaac gctgtctgtg agattgtcgt attctagcct
     7681 ttttagtttt tcgctcatcg acttgatatt gtccgacaca ttttcgtcga tttgcgtttt
     7741 gatcaaagac ttgagcagag acacgttaat caactgttca aattgatcca tattaacgat
     7801 atcaacccga tgcgtatatg gtgcgtaaaa tatatttttt aaccctctta tactttgcac
     7861 tctgcgttaa tacgcgttcg tgtacagacg taatcatgtt ttcttttttg gataaaactc
     7921 ctactgagtt tgacctcata ttagaccctc acaagttgca aaacgtggca ttttttacca
     7981 atgaagaatt taaagttatt ttaaaaaatt tcatcacaga tttaaagaag aaccaaaaat
     8041 taaattattt caacagttta atcgaccagt taatcaacgt gtacacagac gcgtcggcaa
     8101 aaaacacgca gcccgacgtg ttggctaaaa ttattaaatc aacttgtgtt atagtcacgg
     8161 atttgccgtc caacgtgttc ctcaaaaagt tgaagaccaa caagtttacg gacactatta
     8221 attatttgat tttgccccac ttcattttgt gggatcacaa ttttgttata tttttaaaca
     8281 aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     8341 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     8401 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     8461 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     8521 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     8581 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     8641 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     8701 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     8761 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     8821 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     8881 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     8941 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     9001 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     9061 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     9121 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     9181 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     9241 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     9301 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     9361 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     9421 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     9481 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     9541 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     9601 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     9661 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     9721 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     9781 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     9841 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     9901 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     9961 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
    10021 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
    10081 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
    10141 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
    10201 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
    10261 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
    10321 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
    10381 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
    10441 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
    10501 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
    10561 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
    10621 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
    10681 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
    10741 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
    10801 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
    10861 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattc
    10921 gagctcggta cccggggatc ctctagagtc gacgctcgcg cgacttggtt tgccattctt
    10981 tagcgcgcgt cgcgtcacac agcttggcca caatgtggtt tttgtcaaac gaagattcta
    11041 tgacgtgttt aaagtttagg tcgagtaaag cgcaaatctt ttttaaccct agaaagatag
    11101 tctgcgtaaa attgacgcat gcattcttga aatattgctc tctctttcta aatagcgcga
    11161 atccgtcgct gtgcatttag gacatctcag tcgccgcttg gagctcccgt gaggcgtgct
    11221 tgtcaatgcg gtaagtgtca ctgattttga actataacga ccgcgtgagt caaaatgacg
    11281 catgattatc ttttacgtga cttttaagat ttaactcata cgataattat attgttattt
    11341 catgttctac ttacgtgata acttattata tatatatttt cttgttatag atatcgtgac
    11401 taatatataa taaaatgggt agttctttag acgatgagca tatcctctct gctcttctgc
    11461 aaagcgatga cgagcttgtt ggtgaggatt ctgacagtga aatatcagat cacgtaagtg
    11521 aagatgacgt ccagagcgat acagaagaag cgtttataga tgaggtacat gaagtgcagc
    11581 caacgtcaag cggtagtgaa atattagacg aacaaaatgt tattgaacaa ccaggttctt
    11641 cattggcttc taacagaatc ttgaccttgc cacagaggac tattagaggt aagaataaac
    11701 attgttggtc aacttcaaag tccacgaggc gtagccgagt ctctgcactg aacattgtca
    11761 gatccgagat cggccggcct agaagcttta agatacattg atgagtttgg acaaaccaca
    11821 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
    11881 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
    11941 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
    12001 atggctgatt atgatctaga gtcctacagg aacaggtggt ggcggccctc ggtgcgctcg
    12061 tactgctcca cgatggtgta gtcctcgttg tgggaggtga tgtccagctt ggagtccacg
    12121 tagtagtagc cgggcagctg cacgggcttc ttggccatgt agatggactt gaactccacc
    12181 aggtagtggc cgccgtcctt cagcttcagg gccttgtgga tctcgccctt cagcacgccg
    12241 tcgcgggggt acaggcgctc ggtggaggcc tcccagccca tggtcttctt ctgcattacg
    12301 gggccgtcgg aggggaagtt cacgccgatg aacttcacct tgtagatgaa gcagccgtcc
    12361 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
    12421 tcccacttga agccctcggg gaaggacagc ttcttgtagt cggggatgtc ggcggggtgc
    12481 ttcacgtaca ccttggagcc gtactggaac tggggggaca ggatgtccca ggcgaagggc
    12541 agggggccgc ccttggtcac cttcagcttc acggtgttgt ggccctcgta ggggcggccc
    12601 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg tgccctccat gcgcaccttg
    12661 aagcgcatga actccttgat gacgttcttg gaggagcgca ccatcaccag agacaggttg
    12721 cggcggcggt tggatggcgt gggcgcgttg gcgttgttgg accggctcat gttgtgtcgc
    12781 tgtaacagat gctgttcaac tgtgtttacc agatcgttgc gggctgtatt tataggcgcg
    12841 ataagcggga cgggcgcctc gtgtccggtc acgcgcatga gataacgcgc ggctgatatg
    12901 gaggcgcgtc ctgttccgat aaggagttgc gtccggctgc ggttagcaac acaggaagct
    12961 ggcgtcctgt cacgataaga caacactcgt ccggtccgat aatgtgattc gtacgtgaca
    13021 ggacgcgacc cgataaggcc ggcctacgtg actgccgaca cgtacttttt tgcactgcaa
    13081 aaaggttcaa tgtgtggtag tgtatttgga gcgtatacaa cggtgtagac tatttatgta
    13141 aaatagtcta cgaaacgtag agtttgtact atgtatgggc ccgcgtgcaa aagcgtgttt
    13201 ttttgcagtg caaaaaagtt ggtggtgggg aggccaccga gtatgggcgc gccccggccg
    13261 ttaactcgaa tcgctatcca agccagctcc attgtcggca tcgtgctcat tctattgacg
    13321 ctggcagatt tggttttggc gctatgggac ccgttcggtt acaacaacat gtttccgcgc
    13381 gagtttcccg acgacatgtc gcgcacgttc ctgactgcgt actttgagag tttcgacaac
    13441 accacgtcca gagaaatcat agagtttatg cccgagttct tttcggaaat ggtcgaaacg
    13501 gacgatgacg ccacgtttga atctctattt catttattag attatgtggc atctttagaa
    13561 gttaattccg acggccaaat gttaaacttg gaggagggtg atgaaattga ggattttgac
    13621 gaatctactt tggtggggca agcgttagcc actagctcgc tatacactcg catggagttt
    13681 atgcagtaca cgtttaggca aaacacacta ttgtctatga acaaagaaaa caacaatttt
    13741 aatcaaataa tcatgggttt atttgcaaca aacacaattg tggcgtttac agcatttgtt
    13801 atacacacag aactcatatt ttttatattt ttcgtaatct tcctaatgat cacattttat
    13861 tacataatca aagaatcgta cgaatattat aaaacaattg atttgttatt ttaaaaacga
    13921 ttcattctag ttaattaatt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
    13981 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
    14041 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
    14101 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcgg
//
