LOCUS       sequence-211079-        9173 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pX459 V2.0 (Plasmid #62988) with the N497A, R661A, Q695A, and Q926A 
            mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9173)
  AUTHORS   Kato-Inui T, Takahashi G, Hsu S, Miyaoka Y
  TITLE     Clustered regularly interspaced short palindromic repeats 
            (CRISPR)/CRISPR-associated protein 9 with improved proof-reading 
            enhances homology-directed repair.
  JOURNAL   Nucleic Acids Res. 2018 May 18;46(9):4677-4688. doi: 
            10.1093/nar/gky264.
  PUBMED    29672770
REFERENCE   2  (bases 1 to 9173)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2018-05-18- 18"; volume: "46"; issue: 
            "9"; pages: "4677-4688"
COMMENT     Sequence Label: pX459V2.0-SpCas9-HF1
FEATURES             Location/Qualifiers
     source          1..9173
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          130..357
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      369..378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             378..443
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             450..470
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(495..1979,1980..1982,1983..2471,2472..2474,2475..2573,
                     2574..2576,2577..3266,3267..3269,3270..4595)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Kleinstiver et al., 2016)"
                     /label=SpCas9-HF1
                     /note="carries the mutations N497A, R661A, Q695A, and 
                     Q926A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             495..4595
                     /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
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4596..4643
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4659..4712
                     /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             4713..5312
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4713..4732)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5209..5229
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5340..5357)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5346..5553
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5562..5702
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5562..5691
                     /label=AAV2 ITR
     rep_origin      5777..6232
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5864..5883)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6074..6095
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6249..6268)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6368..6390
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6428..6446)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6514..6618
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6619..6687,6688..7479)
                     /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(6837..6856)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7650..8238
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8139..8158
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8300..8548
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8300..8320
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8471..8490
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8567..8642
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8739..9024
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggggggggg gggggggggg gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
       61 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
      121 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
      181 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
      241 ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt
      301 ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt
      361 ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat catgatattg
      421 attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg
      481 gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
      541 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
      601 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
      661 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
      721 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
      781 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
      841 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
      901 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
      961 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     1021 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     1081 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     1141 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     1201 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     1261 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     1321 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     1381 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     1441 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     1501 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     1561 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     1621 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     1681 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     1741 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     1801 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     1861 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     1921 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgaccg
     1981 ccttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     2041 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     2101 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     2161 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     2221 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     2281 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     2341 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     2401 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     2461 ccggctgggg cgcgctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     2521 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atggcgctga
     2581 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     2641 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     2701 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     2761 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     2821 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     2881 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     2941 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     3001 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     3061 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     3121 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     3181 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     3241 tcaagagaca gctggtggaa acccgggcga tcacaaagca cgtggcacag atcctggact
     3301 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     3361 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     3421 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     3481 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     3541 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     3601 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     3661 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     3721 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     3781 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     3841 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     3901 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     3961 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     4021 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     4081 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     4141 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     4201 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     4261 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     4321 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     4381 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     4441 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     4501 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     4561 agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc acgaaaaagg
     4621 ccggccaggc aaaaaagaaa aaggaattcg gcagtggaga gggcagagga agtctgctaa
     4681 catgcggtga cgtcgaggag aatcctggcc caatgaccga gtacaagccc acggtgcgcc
     4741 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     4801 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     4861 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     4921 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     4981 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5041 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag
     5101 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5161 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5221 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5281 cctggtgcat gacccgcaag cccggtgcct gagaattcta actagagctc gctgatcagc
     5341 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     5401 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     5461 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     5521 ggattgggaa gagaatagca ggcatgctgg ggagcggccg caggaacccc tagtgatgga
     5581 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     5641 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     5701 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     5761 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5821 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     5881 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5941 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     6001 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6061 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     6121 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     6181 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     6241 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     6301 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6361 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     6421 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6481 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6541 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6601 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6661 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6721 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6781 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6841 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6901 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6961 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7021 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7081 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7141 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     7201 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7261 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7321 gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7381 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7441 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7501 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7561 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7621 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7681 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7741 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7801 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7861 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7921 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7981 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     8041 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     8101 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8161 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8221 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8281 tggccttttg ctcacatgtg agggcctatt tcccatgatt ccttcatatt tgcatatacg
     8341 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
     8401 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     8461 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     8521 atatatcttg tggaaaggac gaaacaccgg gtcttcgaga agacctgttt tagagctaga
     8581 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     8641 gcttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc
     8701 gtgcgccaat tctgcagaca aatggctcta gaggtacccg ttacataact tacggtaaat
     8761 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaatagt aacgccaata
     8821 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     8881 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     8941 gcctggcatt gtgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     9001 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     9061 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     9121 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggggggggg ggg
//
