LOCUS       sequence-169587-        9035 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses sgRNA targeting human APC around codon I1417.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9035)
  AUTHORS   Guernet A, Mungamuri SK, Cartier D, Sachidanandam R, Jayaprakash A, 
            Adriouch S, Vezain M, Charbonnier F, Rohkin G, Coutant S, Yao S, 
            Ainani H, Alexandre D, Tournier I, Boyer O, Aaronson SA, Anouar Y, 
            Grumolato L
  TITLE     CRISPR-Barcoding for Intratumor Genetic Heterogeneity Modeling and 
            Functional Analysis of Oncogenic Driver Mutations.
  JOURNAL   Mol Cell. 2016 Aug 4;63(3):526-38. doi: 
            10.1016/j.molcel.2016.06.017. Epub 2016 Jul 21.
  PUBMED    27453044
REFERENCE   2  (bases 1 to 9035)
  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: 
            "10.1016/j.molcel.2016.06.017"; journalName: "Mol Cell"; date: 
            "2016-08-4- 4"; volume: "63"; issue: "3"; pages: "526-38"
COMMENT     Sequence Label: sgAPC(A)
FEATURES             Location/Qualifiers
     source          1..9035
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          29..256
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      268..277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             277..342
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             349..369
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             394..4494
                     /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             394..4494
                     /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             4495..4542
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4558..4611
                     /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             4612..5211
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4612..4631)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(5239..5256)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5245..5452
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5461..5601
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5461..5590
                     /label=AAV2 ITR
     rep_origin      5676..6131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5763..5782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5973..5994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6148..6167)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6267..6289
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6327..6345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6413..6517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6518..6586,6587..7378)
                     /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(6736..6755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7549..8137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8038..8057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8199..8447
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8199..8219
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8370..8389
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8469..8544
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8641..8926
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
       61 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
      121 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagctgagca agaggtaagg
      181 gtttaaggga tggttggttg gtggggtatt aatgtttaat tacctggagc acctgcctga
      241 aatcactttt tttcaggttg gaccggtgcc accatggact ataaggacca cgacggagac
      301 tacaaggatc atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag
      361 cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac
      421 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
      481 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
      541 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
      601 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
      661 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
      721 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
      781 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
      841 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
      901 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
      961 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     1021 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     1081 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     1141 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     1201 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     1261 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     1321 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     1381 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     1441 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     1501 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     1561 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     1621 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     1681 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     1741 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     1801 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     1861 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     1921 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     1981 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     2041 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     2101 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     2161 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     2221 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     2281 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     2341 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     2401 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     2461 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     2521 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     2581 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     2641 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     2701 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     2761 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     2821 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     2881 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     2941 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     3001 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     3061 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     3121 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     3181 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     3241 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     3301 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     3361 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     3421 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     3481 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     3541 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     3601 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     3661 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     3721 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     3781 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     3841 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     3901 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     3961 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     4021 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
     4081 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     4141 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     4201 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     4261 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     4321 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     4381 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
     4441 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg
     4501 ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa aggaattcgg cagtggagag
     4561 ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc aatgaccgag
     4621 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     4681 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     4741 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4801 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4861 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4921 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4981 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     5041 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5101 gcgccccaca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5161 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agaattctaa
     5221 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     5281 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     5341 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     5401 ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc
     5461 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     5521 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     5581 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     5641 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     5701 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     5761 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5821 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5881 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5941 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6001 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     6061 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6121 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6181 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6241 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6301 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     6361 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6421 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6481 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6541 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6601 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6661 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6721 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6781 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6841 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6901 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6961 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7021 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7081 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7141 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7201 tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg
     7261 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7321 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7381 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7441 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7501 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7561 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7621 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7681 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7741 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7801 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7861 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7921 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7981 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     8041 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8101 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8161 acggttcctg gccttttgct ggccttttgc tcacatgtga gggcctattt cccatgattc
     8221 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
     8281 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     8341 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     8401 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgag ggctatctgg
     8461 aagatcacgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     8521 gaaaaagtgg caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat
     8581 aaggctagtc cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaggtacc
     8641 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     8701 gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     8761 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     8821 cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt
     8881 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
     8941 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
     9001 tattttttaa ttattttgtg cagcgatggg ggcgg
//
