LOCUS       sequence-275570-        9007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FRB fused SpCas9 for NanoMEDIC packaging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9007)
  AUTHORS   Gee P, Lung MSY, Okuzaki Y, Sasakawa N, Iguchi T, Makita Y, Hozumi 
            H, Miura Y, Yang LF, Iwasaki M, Wang XH, Waller MA, Shirai N, Abe 
            YO, Fujita Y, Watanabe K, Kagita A, Iwabuchi KA, Yasuda M, Xu H, 
            Noda T, Komano J, Sakurai H, Inukai N, Hotta A
  TITLE     Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and 
            sgRNA to induce therapeutic exon skipping.
  JOURNAL   Nat Commun. 2020 Mar 13;11(1):1334. doi: 10.1038/s41467-020-14957-y.
  PUBMED    32170079
REFERENCE   2  (bases 1 to 9007)
  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: "2020-03-13"; volume: "11(1):1334. doi"; pages: " 
            10.1038/s41467-020-14957-y"
COMMENT     Sequence Label: pHLS-EF1a-FRB-SpCas9-A
FEATURES             Location/Qualifiers
     source          1..9007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    23..47
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      56..65
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             74..352
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             359..4462
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             359..4462
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     RBS             4375..4383
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4469..4516
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4523..4603
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             4619..4639
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(4652..4676)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4841..4860)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4906..4961
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4960..4979)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5377..5394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5548..6136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5628..5647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6307..7098,7099..7167))
                     /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     6930..6949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7168..7272)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7449..7584
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7701..7719
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7829..9007
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8060..8998
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     8955..8975
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 ggaattagcg gtggcggcca tcacaagttt gtacaaaaaa gcaggctgtc gagccgccac
       61 catggcttct agaatcctct ggcatgagat gtggcatgaa ggcctggaag aggcatctcg
      121 tttgtacttt ggggaaagga acgtgaaagg catgtttgag gtgctggagc ccttgcatgc
      181 tatgatggaa cggggccccc agactctgaa ggaaacatcc tttaatcagg cctatggtcg
      241 agatttaatg gaggcccaag agtggtgcag gaagtacatg aaatcaggga atgtcaagga
      301 cctcctccaa gcctgggacc tctattatca tgtgttccga cgaatctcaa agtcgaccat
      361 ggataagaaa tacagcattg gactggacat tgggacaaac tccgtgggat gggccgtgat
      421 tacagacgaa tacaaagtgc cttcaaagaa gttcaaggtg ctgggcaaca ccgatagaca
      481 cagcatcaag aaaaatctga ttggagccct gctgttcgac tccggcgaga cagctgaagc
      541 aactcggctg aaaagaactg ctcggagaag gtatacccgc cgaaagaata ggatctgcta
      601 cctgcaggag attttcagca acgaaatggc caaggtggac gatagtttct ttcaccgcct
      661 ggaggaatca ttcctggtcg aggaagataa gaaacacgag cggcatccca tctttggcaa
      721 cattgtggac gaggtcgctt atcacgaaaa gtaccctacc atctatcatc tgaggaagaa
      781 actggtggac tccacagata aagcagacct gcgcctgatc tatctggccc tggctcacat
      841 gattaagttc cggggccatt ttctgatcga gggggatctg aacccagaca attctgatgt
      901 ggacaagctg ttcatccagc tggtccagac atacaatcag ctgtttgagg aaaaccccat
      961 taatgcatct ggcgtggacg caaaagccat cctgagtgcc agactgtcta agagtcggag
     1021 actggagaac ctgatcgctc agctgccagg ggaaaagaaa aacggcctgt ttgggaatct
     1081 gattgcactg tcactgggac tgactcccaa cttcaagagc aattttgatc tggccgagga
     1141 cgctaaactg cagctgtcca aggacaccta tgacgatgac ctggataacc tgctggctca
     1201 gatcggggat cagtacgcag acctgttcct ggccgctaag aatctgtctg acgccatcct
     1261 gctgagtgat attctgcgcg tgaacaccga gattacaaaa gcccccctgt cagctagcat
     1321 gatcaagaga tatgacgagc accatcagga tctgaccctg ctgaaggctc tggtgaggca
     1381 gcagctgcct gagaagtaca aggaaatctt ctttgatcag tctaagaacg gatacgccgg
     1441 ctatattgac ggcggggcta gtcaggagga gttctacaag tttatcaaac ccattctgga
     1501 gaagatggat ggcacagagg aactgctggt gaaactgaat cgggaagacc tgctgaggaa
     1561 gcagcgcact tttgataacg gaagcatccc tcaccagatt catctgggag agctgcacgc
     1621 aatcctgagg cgccaggaag acttctaccc atttctgaag gataacaggg agaagatcga
     1681 aaaaattctg acattccgca tcccctacta tgtgggccct ctggcaagag gcaacagccg
     1741 gtttgcctgg atgactcgca aatctgagga aacaatcact ccctggaact tcgaggaagt
     1801 ggtcgataag ggcgcttccg cacagtcttt cattgagcgg atgacaaact tcgacaagaa
     1861 cctgccaaac gaaaaagtgc tgcccaagca ctctctgctg tacgagtatt tcacagtcta
     1921 taacgaactg actaaggtga aatacgtcac cgaggggatg agaaagcctg ccttcctgag
     1981 tggagaacag aagaaagcta tcgtggacct gctgtttaaa accaatagga aggtgacagt
     2041 caagcagctg aaagaggact atttcaagaa aattgaatgt ttcgattctg tggagatcag
     2101 tggcgtcgaa gacaggttta acgcctccct ggggacctac cacgatctgc tgaagatcat
     2161 taaggataaa gacttcctgg acaacgagga aaatgaggat atcctggaag acattgtgct
     2221 gaccctgaca ctgtttgagg atagggaaat gatcgaggaa cgcctgaaga cctatgccca
     2281 tctgttcgat gacaaagtga tgaaacagct gaagcgacgg agatacacag gatggggccg
     2341 actgtctcgg aagctgatca atgggattcg cgacaaacag agtggaaaga ccatcctgga
     2401 ctttctgaaa tcagatggct tcgccaaccg gaacttcatg cagctgattc acgatgacag
     2461 cctgacattc aaagaggata tccagaaggc acaggtgtcc gggcagggag actctctgca
     2521 cgagcatatc gcaaacctgg ccggcagccc tgccatcaag aaagggattc tgcagaccgt
     2581 gaaggtggtg gacgagctgg tgaaagtcat gggaagacat aagccagaaa acatcgtgat
     2641 tgagatggcc agggaaaatc agaccacaca gaaaggccag aagaactcaa gggagcgcat
     2701 gaaaagaatc gaggaaggaa ttaaggaact gggcagccag atcctgaaag agcaccccgt
     2761 ggaaaacaca cagctgcaga atgagaagct gtatctgtac tatctgcaga atggacgcga
     2821 tatgtacgtg gaccaggagc tggatattaa ccgactgtcc gattacgacg tggatcatat
     2881 cgtcccacag tcattcctga aagatgacag cattgacaat aaggtgctga cccgctctga
     2941 caaaaaccga ggcaagagtg ataatgtccc ctcagaggaa gtggtcaaga aaatgaagaa
     3001 ctactggagg cagctgctga atgccaaact gatcacacag cgaaagtttg ataacctgac
     3061 taaagctgag cggggaggcc tgagtgaact ggacaaagca ggcttcatta agcgacagct
     3121 ggtggagaca cggcagatca caaagcacgt cgcccagatt ctggattcaa gaatgaacac
     3181 taagtacgat gagaatgaca aactgatcag agaagtgaag gtcattaccc tgaagtcaaa
     3241 actggtgagc gactttcgga aagatttcca gttttataag gtcagagaga tcaacaacta
     3301 ccaccatgct catgacgcat acctgaacgc agtggtcggc acagccctga ttaagaaata
     3361 ccctaaactg gagtccgagt tcgtgtacgg ggactataag gtgtacgatg tcagaaaaat
     3421 gatcgccaag tctgagcagg aaattggcaa agccactgct aagtatttct tttacagtaa
     3481 catcatgaat ttctttaaga ctgagatcac cctggcaaat ggggaaatcc gaaagcggcc
     3541 actgattgag actaacggcg agacaggaga aatcgtgtgg gacaaaggaa gagattttgc
     3601 taccgtgagg aaggtcctga gcatgcccca agtgaatatt gtcaagaaaa cagaggtgca
     3661 gactggggga ttcagtaagg aatcaattct gcctaaacgc aactccgata agctgatcgc
     3721 ccgaaagaaa gactgggacc ccaagaagta tggcgggttc gactccccaa ctgtggctta
     3781 ctctgtcctg gtggtcgcaa aggtggagaa gggaaaaagc aagaaactga aatccgtcaa
     3841 ggaactgctg ggcatcacca ttatggagcg cagctccttc gaaaagaatc ctatcgattt
     3901 tctggaggcc aaaggctata aggaagtgaa gaaagacctg atcatcaagc tgccaaagta
     3961 ctcactgttt gagctggaaa acgggagaaa gaggatgctg gcaagcgccg gggagctgca
     4021 gaaaggaaat gaactggccc tgccctccaa gtacgtgaac ttcctgtatc tggctagcca
     4081 ctacgagaag ctgaaagggt cccctgagga taacgaacag aaacagctgt ttgtggagca
     4141 gcacaagcat tatctggacg agatcattga acagattagc gagttctcca aaagagtgat
     4201 cctggctgac gcaaatctgg ataaggtcct gagcgcatac aacaaacacc gggataagcc
     4261 aatcagagag caggccgaaa atatcattca tctgttcact ctgaccaacc tgggagcccc
     4321 cgcagccttc aagtattttg acactaccat cgatcgcaaa cgatacacaa gcactaagga
     4381 ggtgctggac gctaccctga ttcatcagag cattactggc ctgtatgaaa caaggattga
     4441 cctgtctcag ctgggcggcg actccggaaa aaggccggcg gccacgaaaa aggccggcca
     4501 ggcaaaaaag aaaaagggat cctacccata cgatgttcca gattacgctt acccatacga
     4561 tgttccagat tacgcttacc catacgatgt tccagattac gcttaatccg gagctgaccc
     4621 caagaagaag aggaaggtgt gagacctcta gacccagctt tcttgtacaa agtggtgatc
     4681 tagctaaaac gcggcctcga atcgaattcg attcgaatgt acagatctgg atcctaggct
     4741 taagtatacc tcgagttaaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg
     4801 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
     4861 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     4921 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     4981 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     5041 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     5101 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     5161 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     5221 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     5281 ggagatccct cgacttaatt aaaccggtca gctgcattaa tgaatcggcc aacgcgcggg
     5341 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5401 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5461 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5521 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5581 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5641 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5701 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5761 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5821 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5881 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5941 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6001 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6061 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6121 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6181 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6241 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6301 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6361 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6421 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6481 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6541 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6601 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6661 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6721 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6781 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6841 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6901 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6961 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7021 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7081 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7141 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7201 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7261 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgctagcg tttaaacccg
     7321 ggcgatcgct ctagattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggacta gctttttgca
     7441 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     7501 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     7561 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     7621 attgagatga gcttgcatgc cgacattgat tattgactag tccctaagaa accattctta
     7681 tcatgacatt aacctataaa aataggcgta tcacgaggcc tagactagtg cggccgctgc
     7741 agaagctttg caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac
     7801 cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     7861 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag
     7921 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     7981 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     8041 gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac
     8101 gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga
     8161 tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc
     8221 ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg
     8281 gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg
     8341 atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct
     8401 gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg
     8461 cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc
     8521 tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg
     8581 ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg
     8641 ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc
     8701 acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga
     8761 gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt
     8821 aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga
     8881 agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg
     8941 atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt
     9001 gtcgtga
//
