LOCUS       sequence-218705-       14985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hSpCas9 under vasa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14985)
  AUTHORS   Kandul NP, Liu J, Sanchez C HM, Wu SL, Marshall JM, Akbari OS
  TITLE     Transforming insect population control with precision guided sterile
            males with demonstration in flies.
  JOURNAL   Nat Commun. 2019 Jan 8;10(1):84. doi: 10.1038/s41467-018-07964-7.
  PUBMED    30622266
REFERENCE   2  (bases 1 to 14985)
  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: "Nat 
            Commun."; date: "2019-01-8"; volume: "10(1):84. doi"; pages: " 
            10.1038/s41467-018-07964-7"
COMMENT     Sequence Label: vas-Cas9.874Z
FEATURES             Location/Qualifiers
     source          1..14985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(95..683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(175..194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(854..1645,1646..1714))
                     /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     1477..1496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1715..1819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1887..1905
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1943..1965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2065..2084
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2278..2300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2292..2309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2293..2309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   3013..3075
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4109..4178
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     CDS             6606..6671
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6678..6698
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6723..10823
                     /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             6723..10823
                     /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             10824..10871
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10872..10925
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(10926..10928,10929..10931,10932..11645)
                     /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(11232..11251)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     11579..11600
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(12880..12882,12883..12885,12886..13560)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(13063..13083)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     13281..13300
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     13475..13498
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    13674..13795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13711..13730)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13765..13784
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   14463..14497
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(14678..14694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14678..14694)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14691..14713)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14702..14718
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(14726..14732,14733..14750,14751..14756))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14771..14792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14909..14926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
       61 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
      541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
      841 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
      961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1861 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1921 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1981 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2041 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2101 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2161 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2221 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2281 agtcacgacg ttgtaaaacg acggccagtg ccaagctttg tttaaaaata taacaaaatt
     2341 gtgatcccac aaaatgaagt ggggcaaaat caaataatta atagtgtccg taaacttgtt
     2401 ggtcttcaac tttttgagga acacgttgga cggcaaatcc gtgactataa cacaagttga
     2461 tttaataatt ttagccaaca cgtcgggctg cgtgtttttt gccgacgcgt ctgtgtacac
     2521 gttgattaac tggtcgatta aactgttgaa ataatttaat ttttggttct tctttaaatc
     2581 tgtgatgaaa ttttttaaaa taactttaaa ttcttcattg gtaaaaaatg ccacgttttg
     2641 caacttgtga gggtctaata tgaggtcaaa ctcagtagga gttttatcca aaaaagaaaa
     2701 catgattacg tctgtacacg aacgcgtatt aacgcagagt gcaaagtata agagggttaa
     2761 aaaatatatt ttacgcacca tatacgcatc gggttgatat cgttaatatg gatcaatttg
     2821 aacagttgat taacgtgtct ctgctcaagt ctttgatcaa aacgcaaatc gacgaaaatg
     2881 tgtcggacaa tatcaagtcg atgagcgaaa aactaaaaag gctagaatac gacaatctca
     2941 cagacagcgt tgagatatac ggtattcacg acagcaggct gaataataaa aaaattagaa
     3001 actattattt aaccctagaa agataatcat attgtgacgt acgttaaaga taatcatgcg
     3061 taaaattgac gcatgtgttt tatcggtctg tatatcgagg tttatttatt aatttgaata
     3121 gatattaagt tttattatat ttacacttac atactaataa taaattcaac aaacaattta
     3181 tttatgttta tttatttatt aaaaaaaaac aaaaactcaa aatttcttct ataaagtaac
     3241 aaaactttta aacattctct cttttacaaa aataaactta ttttgtactt taaaaacagt
     3301 catgttgtat tataaaataa gtaattagct taacttatac ataatagaaa caaattatac
     3361 ttattagtca gtcagaaaca actttggcac atatcaatat tatgctctcg acaaataact
     3421 tttttgcatt ttttgcacga tgcatttgcc tttcgcctta ttttagaggg gcagtaagta
     3481 cagtaagtac gttttttcat tactggctct tcagtactgt catctgatgt accaggcact
     3541 tcatttggca aaatattaga gatattatcg cgcaaatatc tcttcaaagt aggagcttct
     3601 aaacgcttac gcataaacga tgacgtcagg ctcatgtaaa ggtttctcat aaattttttg
     3661 cgactttgaa ccttttctcc cttgctactg acattatggc tgtatataat aaaagaattt
     3721 atgcaggcaa tgtttatcat tccgtacaat aatgccatag gccacctatt cgtcttccta
     3781 ctgcaggtca tcacagaaca catttggtct agcgtgtcca ctccgccttt agtttgatta
     3841 taatacataa ccatttgcgg tttaccggta ctttcgttga tagaagcatc ctcatcacaa
     3901 gatgataata agtataccat cttagctggc ttcggtttat atgagacgag agtaaggggt
     3961 ccgtcaaaac aaaacatcga tgttcccact ggcctggagc gactgttttt cagtacttcc
     4021 ggtatctcgc gtttgtttga tcgcacggtt cccacaatgg ttaattcgag ctcgcccggg
     4081 gtcctaggtc gacgatgtag gtcacggtct cgaagccgcg gtgcgggtgc cagggcgtgc
     4141 ccttgggctc cccgggcgcg tactccacct cacccatctg gtccatcatg atgaacgggt
     4201 cgaggtggcg gtagttgatc ccggcgaacg cgcggcgcac cgggaagccc tcgccctcga
     4261 aaccgctggg cgcggtggtc acggtgagca cgggacgtgc gacggcgtcg gcgggtgcgg
     4321 atacgcgggg cagcgtcagc gggttctcga cggtcacggc gggcatgtcg acgcctgcag
     4381 ctggttgtag gtgcagttgc gcttcttgca cttggcgcac ttgaggagat cggtcttggt
     4441 gccctgcacg gtggccagct gggcatcgtt gatggcctcc ttgacgaact tctcgcgcag
     4501 cttcttcatc tcgtcgctgg ccatctcctc cggcgtcatt ttggccagct gcttggcagt
     4561 gacggcgccg cacataaagt tgccgcgcaa tccaggattc ttggggtcct tcagattggc
     4621 cacgcgtgac ctaatgcgat tcttgtactt catatccgtg ttattgaact cggagtaaat
     4681 ggcatcctcc agttcggcgg ccatttcctc cggctcaccg catccctcgg gcacttcgcc
     4741 aatcttcagt gcggtggcca gcatttcgcg gcatttgatg cggaccgcat ctgtcatgcc
     4801 accggaggga aacgaggtct gtgacgagga tgtcgagcct ttcttctctt tatccttgga
     4861 tgagctggat gaactggact tatccttgcc ggagattgac gacgacgact tggcggcact
     4921 ggtggacttc gaggcactgc tgttgttgga ggagccctcc ttagcggagc tgttgttcgg
     4981 tgtggttggc gccgggctgg ccaggaagcg tttccagttc ttgatcagcg tcttggccag
     5041 agcgatcacc tcgtcgtcct tgctactctt gcgcagctcg tttacggtca tgccgatgcg
     5101 cgttttggtc agaatgtcga gattgatgtt aagcgtttgc agggccttca gcaggtccag
     5161 agcctgatcc tgtccctgta cggtggatat atgtatattt gtgacatgat gcgcggcacg
     5221 ttagctaatc ccgatactta ctgtgccgtc gctggccatc ttgctcatct tcttttggat
     5281 tcgaaacact tcctcctcca cgctcatttt ggcccactta atttgatcag aggttaggtt
     5341 aactggcgtc ggctgtcggc tttaatcggt gctgcgggtc tgcgacagtt gtttatgcgg
     5401 aagccgggct atttcttttt ttccctccaa taaatgttgt ttactatttt tttcgtttct
     5461 gtattcgcag aacaagttga cagaacaccc taacgcgttt tagtgttacc gggcggtggc
     5521 ttgtcaagaa ataccttccc atggctaata caattcccaa ctggaagtag tttgggtttc
     5581 agaagtcaac agtgttaccg tattttaatg tacatctggc ttttaagcga gtacacacac
     5641 atagcattgc gatgtttttt aaacttgcac tgcttgcaag caggggtcca ctgtatttca
     5701 atgaaagaac agcgtgttgg gcgtttttga attatactaa ttaccttcaa taacaattcc
     5761 ctatatatca cttagtttta ataaataaat cgtttgagtg tggtctagag atgatttatt
     5821 atgatgaagt gcggttgttg gggacaagcg ggatacgtct gcgggtcaaa tcggcggtgg
     5881 gacatagtga gagggcaacg gcgaacaatc accaacgcgc cctggtacgc gagaaatgtt
     5941 gatatcgcac atgacctcag catccacctt cagtttgaga acaaattaaa tcaggcacat
     6001 accagacaac caggagaaca ctgaggaggc gtcatcaaag atatctttgg acacgtggca
     6061 taaacaagcc aaactattat ctaatcaata tttttataca aattttgtat gtttccatgt
     6121 tatcttcgtt gctcctgtta gttactacgt ttcatagccc ttaaattggt tgcttaacgt
     6181 aaaataaaat tatattgtat aaaaataaag aggaattcac ctataacaaa cgaaaaagta
     6241 aataaataag aacttcccca tgttaaaaat gccaccacca tctttacata taagtatgta
     6301 catatgaatg aatcacttag gttgcttgaa tattacaatt tcattaatag ctaaatcaca
     6361 tttgatgtgt tagtggaaaa cggctatata tataaattat cgaaattgtg aatatcgaat
     6421 tgcgatagca caatgggaaa ttccaccact agatttttgg tacttttaac agatcctttt
     6481 cggttttgcg ttgcgcgaag tgatctgaac ttatcaaagt ttgtaaggta atacataaag
     6541 tgaaaaagaa ttaatttgct cttgaaaggc aggccaaatt aaaaaaaaaa tatcaattcg
     6601 agatggacta taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg
     6661 atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
     6721 ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     6781 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     6841 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     6901 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     6961 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     7021 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     7081 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     7141 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     7201 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     7261 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     7321 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     7381 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     7441 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     7501 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     7561 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     7621 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     7681 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     7741 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     7801 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     7861 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     7921 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     7981 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     8041 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     8101 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     8161 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     8221 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     8281 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     8341 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     8401 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     8461 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     8521 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     8581 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     8641 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     8701 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     8761 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     8821 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     8881 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     8941 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     9001 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     9061 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     9121 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     9181 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     9241 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     9301 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     9361 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     9421 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     9481 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     9541 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     9601 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     9661 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     9721 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     9781 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     9841 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     9901 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     9961 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
    10021 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
    10081 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
    10141 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
    10201 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
    10261 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
    10321 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
    10381 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
    10441 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
    10501 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
    10561 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
    10621 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
    10681 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
    10741 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
    10801 acctgtctca gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa
    10861 aaaagaaaaa ggagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg
    10921 gccctatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
    10981 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
    11041 cctacggcaa gctgaccctg aagttcatct gcaccaccag caagctgccc gtgccctggc
    11101 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
    11161 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
    11221 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
    11281 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
    11341 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
    11401 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
    11461 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
    11521 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
    11581 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
    11641 agtaattaat taactagaat gaatcgtttt taaaataaca aatcaattgt tttataatat
    11701 tcgtacgatt ctttgattat gtaataaaat gtgatcatta ggaagattac gaaaaatata
    11761 aaaaatatga gttctgtgtg tataacaaat gctgtaaacg ccacaattgt gtttgttgca
    11821 aataaaccca tgattatttg attaaaattg ttgttttctt tgttcataga caatagtgtg
    11881 ttttgcctaa acgtgtactg cataaactcc atgcgagtgt atagcgagct agtggctaac
    11941 gcttgcccca ccaaagtaga ttcgtcaaaa tcctcaattt catcaccctc ctccaagttt
    12001 aacatttggc cgtcggaatt aacttctaaa gatgccacat aatctaataa atgaaataga
    12061 gattcaaacg tggcgtcatc gtccgtttcg accatttccg aaaagaactc gggcataaac
    12121 tctatgattt ctctggacgt ggtgttgtcg aaactctcaa agtacgcagt caggaacgtg
    12181 cgcgacatgt cgtcgggaaa ctcgcgcgga aacatgttgt tgtaaccgaa cgggtcccat
    12241 agcgccaaaa ccaaatctgc cagcgtcaat agaatgagca cgatgccgac aatggagctg
    12301 gcttggatag cgattcgagt taacggccgg ggcgcgccca tactcggtgg cctccccacc
    12361 accaactttt ttgcactgca aaaaaacacg cttttgcacg cgggcccata catagtacaa
    12421 actctacgtt tcgtagacta ttttacataa atagtctaca ccgttgtata cgctccaaat
    12481 acactaccac acattgaacc tttttgcagt gcaaaaaagt acgtgtcggc agtcacgtag
    12541 gccggcctta tcgggtcgcg tcctgtcacg tacgaatcac attatcggac cggacgagtg
    12601 ttgtcttatc gtgacaggac gccagcttcc tgtgttgcta accgcagccg gacgcaactc
    12661 cttatcggaa caggacgcgc ctccatatca gccgcgcgtt atctcatgcg cgtgaccgga
    12721 cacgaggcgc ccgtcccgct tatcgcgcct ataaatacag cccgcaacga tctggtaaac
    12781 acagttgaac agcatctgtt acagcgacac aacatgagcc ggtccaacaa cgccaacgcg
    12841 cccacgccat ccaaccgccg ccgcaacctg tctctggtga tggtgcgctc ctccaagaac
    12901 gtcatcaagg agttcatgcg cttcaaggtg cgcatggagg gcaccgtgaa cggccacgag
    12961 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gccacaacac cgtgaagctg
    13021 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttccag
    13081 tacggctcca aggtgtacgt gaagcacccc gccgacatcc ccgactacaa gaagctgtcc
    13141 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
    13201 gtgacccagg actcctccct gcaggacggc tgcttcatct acaaggtgaa gttcatcggc
    13261 gtgaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
    13321 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacaa ggccctgaag
    13381 ctgaaggacg gcggccacta cctggtggag ttcaagtcca tctacatggc caagaagccc
    13441 gtgcagctgc ccggctacta ctacgtggac tccaagctgg acatcacctc ccacaacgag
    13501 gactacacca tcgtggagca gtacgagcgc accgagggcc gccaccacct gttcctgtag
    13561 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
    13621 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
    13681 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
    13741 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaagct
    13801 tctaggccgg ccgatctcgg atctgacaat gttcagtgca gagactcggc tacgcctcgt
    13861 ggactttgaa gttgaccaac aatgtttatt cttacctcta atagtcctct gtggcaaggt
    13921 caagattctg ttagaagcca atgaagaacc tggttgttca ataacatttt gttcgtctaa
    13981 tatttcacta ccgcttgacg ttggctgcac ttcatgtacc tcatctataa acgcttcttc
    14041 tgtatcgctc tggacgtcat cttcacttac gtgatctgat atttcactgt cagaatcctc
    14101 accaacaagc tcgtcatcgc tttgcagaag agcagagagg atatgctcat cgtctaaaga
    14161 actacccatt ttattatata ttagtcacga tatctataac aagaaaatat atatataata
    14221 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
    14281 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
    14341 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
    14401 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
    14461 tgcatgcgtc aattttacgc agactatctt tctagggtta aaaaagattt gcgctttact
    14521 cgacctaaac tttaaacacg tcatagaatc ttcgtttgac aaaaaccaca ttgtggccaa
    14581 gctgtgtgac gcgacgcgcg ctaaagaatg gcaaaccaag tcgcgcgagc gtcgactcta
    14641 gaggatcccc gggtaccgag ctcgaattcg taatcatgtc atagctgttt cctgtgtgaa
    14701 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    14761 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    14821 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    14881 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    14941 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggt
//
