LOCUS       sequence-218703-       14636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hSpCas9 under Ubiquitin-63E promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14636)
  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 14636)
  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: Ubiq-Cas9.874W
FEATURES             Location/Qualifiers
     source          1..14636
                     /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             6257..6322
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6329..6349
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6374..10474
                     /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             6374..10474
                     /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             10475..10522
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10523..10576
                     /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(10577..10579,10580..10582,10583..11296)
                     /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(10883..10902)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     11230..11251
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(12531..12533,12534..12536,12537..13211)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(12714..12734)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     12932..12951
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     13126..13149
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    13325..13446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13362..13381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13416..13435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   14114..14148
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(14329..14345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14329..14345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14342..14364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14353..14369
                     /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(14377..14383,14384..14401,14402..14407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14422..14443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14560..14577)
                     /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 acgcggccgc
     4381 cgcgcagatc gccgatgggc gtggcgccgg agggcacctt caccttgccg tacggcttgc
     4441 tgttcttcgc gttcaaaatc tccagctcca ttttgctttc ggtgcgcttg caatcagtac
     4501 tgtccaaaat cgaaaatcgc cgaaccgtag tgtgaccgtg cggggctctg cgaaaataaa
     4561 cttttttagg tatatggcca cacacgggga aagcacagtg gattatatgt tttaatatta
     4621 taatatgcag gttttcatta cttatccaga tgtaagccca cttaaagcga tttaacaatt
     4681 atttgccgaa agagtataaa caaatttcac ttaaaaatgg attaagaaaa gcttgtgtaa
     4741 gattatgcgc agcgttgcca gatagctcca tttaaaacac ttcaaaaaca ataagttttg
     4801 aaaatatata cataaatagc agtcgttgcc gcaacgctca acacatcaca cttttaaaac
     4861 accctttacc tacacagaat tactttttaa atttccagtc aagctgcgag tttcaaaatt
     4921 atagccggta gagaagacag tgctatttca aaagcaaact aacaagggtc ttaaattcca
     4981 aaacaccaat cctaacaagc cttggacttt tgtaagttta gatcaaaggt ggcattgcat
     5041 tcaatgtcat ggtaagaagt aggtcgtcta ggtagaaatc ctcattcagc cggtcaagtc
     5101 agtacgagaa aggtctcaat ttgaaattgt cttaaaaata ttttattgtt ttgtactgtg
     5161 gtgagtttaa acgaaaaaca caaaaaaaaa gtgatacaca gaaatcataa aaaattttaa
     5221 tacaaggtat tcgtacgtat caaaaacatt tcggcacaat tttttttctc tgtactaaag
     5281 tgttacgaac actacggtat tttttagtga ttttcaacgg acaccgaagg tatataaaca
     5341 gcgttcgcga acggtcgcct tcaaaaccaa ttgacatttg cagcagcaag tacaagtaga
     5401 aagtaaagcg caatcagcga aaaatttata cttaattgtt ggtgattaaa gtacaattaa
     5461 aagaacattc tcgaaagtca caagaaacgt aagtttttaa ctcgctgtta ccaattagta
     5521 ataagagcaa caagacgttg agtaatttca agaaaaactg catttcaagg tctttgttcg
     5581 gccatttttt ttttattcaa cgctctacgt aattacaaaa taagaaattg gcagccacgc
     5641 atcttgtttt cccaatgaat tggcatcaaa acgcaaacaa atctataaat aaaacttgcg
     5701 tgttgatttt cgccaagatt tattggcaaa ttgtgaaatt cgcagtgacg catttgaaaa
     5761 ttcgagaaat cacgaacgca ctcgagcatt tgtgtgcatg ttattagtta gttgtttagt
     5821 taattgaagt attttaccaa cgaaatccac ttatttttag ctgaaataga gtaggttgct
     5881 taaacaaagc cacgtctgaa aatttcttat tgcttgtagt tgtgacgtca ccatatacac
     5941 acaaaataat gtgtatgcat gcatttcagc tgtgtatata tacatgcaca cactcgcaac
     6001 acgaaaacga tgacgaagca acggaacaaa ggtttctcaa ctaccctttg ttccctgttt
     6061 cttcgctttc ctttgttcca atattcgtag agggttaata ggggtttctc aacaaagttg
     6121 gcgtcgataa ataagtttcc catttttatt ccccagccag gaagttagtt tcaatagttt
     6181 tgtaatttca acgaaactca tttgatttcg tactaatttt ccacatctct attttgaccc
     6241 gcagaattta aatatggact ataaggacca cgacggagac tacaaggatc atgatattga
     6301 ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg
     6361 agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     6421 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     6481 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     6541 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     6601 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     6661 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     6721 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     6781 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     6841 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     6901 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     6961 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     7021 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     7081 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     7141 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     7201 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     7261 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     7321 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     7381 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     7441 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     7501 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     7561 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     7621 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     7681 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     7741 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     7801 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     7861 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     7921 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     7981 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     8041 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     8101 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     8161 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     8221 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     8281 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     8341 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     8401 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     8461 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     8521 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     8581 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     8641 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     8701 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     8761 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     8821 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     8881 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     8941 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     9001 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     9061 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     9121 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     9181 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     9241 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     9301 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     9361 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     9421 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     9481 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     9541 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     9601 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     9661 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     9721 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     9781 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     9841 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     9901 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     9961 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
    10021 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
    10081 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
    10141 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
    10201 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
    10261 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
    10321 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
    10381 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
    10441 gacacggatc gacctgtctc agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc
    10501 cggccaggca aaaaagaaaa aggagggcag aggaagtctt ctaacatgcg gtgacgtgga
    10561 ggagaatccc ggccctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
    10621 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
    10681 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccacca gcaagctgcc
    10741 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
    10801 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
    10861 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
    10921 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
    10981 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
    11041 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
    11101 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
    11161 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
    11221 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
    11281 cgagctgtac aagtaattaa ttaactagaa tgaatcgttt ttaaaataac aaatcaattg
    11341 ttttataata ttcgtacgat tctttgatta tgtaataaaa tgtgatcatt aggaagatta
    11401 cgaaaaatat aaaaaatatg agttctgtgt gtataacaaa tgctgtaaac gccacaattg
    11461 tgtttgttgc aaataaaccc atgattattt gattaaaatt gttgttttct ttgttcatag
    11521 acaatagtgt gttttgccta aacgtgtact gcataaactc catgcgagtg tatagcgagc
    11581 tagtggctaa cgcttgcccc accaaagtag attcgtcaaa atcctcaatt tcatcaccct
    11641 cctccaagtt taacatttgg ccgtcggaat taacttctaa agatgccaca taatctaata
    11701 aatgaaatag agattcaaac gtggcgtcat cgtccgtttc gaccatttcc gaaaagaact
    11761 cgggcataaa ctctatgatt tctctggacg tggtgttgtc gaaactctca aagtacgcag
    11821 tcaggaacgt gcgcgacatg tcgtcgggaa actcgcgcgg aaacatgttg ttgtaaccga
    11881 acgggtccca tagcgccaaa accaaatctg ccagcgtcaa tagaatgagc acgatgccga
    11941 caatggagct ggcttggata gcgattcgag ttaacggccg gggcgcgccc atactcggtg
    12001 gcctccccac caccaacttt tttgcactgc aaaaaaacac gcttttgcac gcgggcccat
    12061 acatagtaca aactctacgt ttcgtagact attttacata aatagtctac accgttgtat
    12121 acgctccaaa tacactacca cacattgaac ctttttgcag tgcaaaaaag tacgtgtcgg
    12181 cagtcacgta ggccggcctt atcgggtcgc gtcctgtcac gtacgaatca cattatcgga
    12241 ccggacgagt gttgtcttat cgtgacagga cgccagcttc ctgtgttgct aaccgcagcc
    12301 ggacgcaact ccttatcgga acaggacgcg cctccatatc agccgcgcgt tatctcatgc
    12361 gcgtgaccgg acacgaggcg cccgtcccgc ttatcgcgcc tataaataca gcccgcaacg
    12421 atctggtaaa cacagttgaa cagcatctgt tacagcgaca caacatgagc cggtccaaca
    12481 acgccaacgc gcccacgcca tccaaccgcc gccgcaacct gtctctggtg atggtgcgct
    12541 cctccaagaa cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggcaccgtga
    12601 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggccacaaca
    12661 ccgtgaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
    12721 cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca
    12781 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
    12841 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg ctgcttcatc tacaaggtga
    12901 agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
    12961 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca
    13021 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagtcc atctacatgg
    13081 ccaagaagcc cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct
    13141 cccacaacga ggactacacc atcgtggagc agtacgagcg caccgagggc cgccaccacc
    13201 tgttcctgta ggactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
    13261 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
    13321 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
    13381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
    13441 atcttaaagc ttctaggccg gccgatctcg gatctgacaa tgttcagtgc agagactcgg
    13501 ctacgcctcg tggactttga agttgaccaa caatgtttat tcttacctct aatagtcctc
    13561 tgtggcaagg tcaagattct gttagaagcc aatgaagaac ctggttgttc aataacattt
    13621 tgttcgtcta atatttcact accgcttgac gttggctgca cttcatgtac ctcatctata
    13681 aacgcttctt ctgtatcgct ctggacgtca tcttcactta cgtgatctga tatttcactg
    13741 tcagaatcct caccaacaag ctcgtcatcg ctttgcagaa gagcagagag gatatgctca
    13801 tcgtctaaag aactacccat tttattatat attagtcacg atatctataa caagaaaata
    13861 tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt atcgtatgag
    13921 ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac gcggtcgtta
    13981 tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga gctccaagcg
    14041 gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa
    14101 tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt aaaaaagatt
    14161 tgcgctttac tcgacctaaa ctttaaacac gtcatagaat cttcgtttga caaaaaccac
    14221 attgtggcca agctgtgtga cgcgacgcgc gctaaagaat ggcaaaccaa gtcgcgcgag
    14281 cgtcgactct agaggatccc cgggtaccga gctcgaattc gtaatcatgt catagctgtt
    14341 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
    14401 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
    14461 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
    14521 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
    14581 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggt
//
