LOCUS       sequence-171512-        9039 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 9039)
  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 9039)
  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(B)
FEATURES             Location/Qualifiers
     source          1..9039
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          30..257
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      269..278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             278..343
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             350..370
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             395..4495
                     /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             395..4495
                     /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             4496..4543
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4559..4612
                     /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             4613..5212
                     /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(4613..4632)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(5240..5257)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5246..5453
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5462..5602
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5462..5591
                     /label=AAV2 ITR
     rep_origin      5677..6132
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5764..5783)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5974..5995
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6149..6168)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6268..6290
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6328..6346)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6414..6518
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6519..6587,6588..7379)
                     /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(6737..6756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7550..8138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8039..8058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8200..8448
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8200..8220
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8371..8390
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8470..8545
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8642..8927
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc
       61 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
      121 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag
      181 ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg
      241 aaatcacttt ttttcaggtt ggaccggtgc caccatggac tataaggacc acgacggaga
      301 ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa
      361 gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga
      421 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
      481 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
      541 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
      601 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
      661 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
      721 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
      781 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
      841 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
      901 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
      961 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     1021 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     1081 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     1141 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     1201 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     1261 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     1321 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     1381 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     1441 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     1501 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     1561 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     1621 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     1681 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     1741 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     1801 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     1861 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     1921 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     1981 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     2041 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     2101 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     2161 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     2221 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     2281 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     2341 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     2401 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     2461 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     2521 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     2581 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     2641 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     2701 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     2761 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     2821 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     2881 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     2941 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     3001 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     3061 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     3121 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     3181 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     3241 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     3301 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     3361 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     3421 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     3481 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     3541 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     3601 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     3661 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     3721 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     3781 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     3841 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     3901 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     3961 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     4021 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     4081 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     4141 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     4201 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     4261 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     4321 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     4381 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     4441 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaaaag
     4501 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggaattcg gcagtggaga
     4561 gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc caatgaccga
     4621 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     4681 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     4741 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4801 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4861 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4921 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4981 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5041 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5101 cgcgccccac aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5161 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagaattcta
     5221 actagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     5281 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     5341 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     5401 gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagcggccg
     5461 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     5521 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     5581 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     5641 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     5701 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     5761 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5821 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5881 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5941 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     6001 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     6061 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     6121 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6181 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     6241 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     6301 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     6361 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     6421 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6481 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6541 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6601 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6661 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6721 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6781 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6841 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6901 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6961 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     7021 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     7081 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     7141 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     7201 ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg
     7261 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7321 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7381 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7441 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7501 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7561 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7621 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7681 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7741 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7801 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7861 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7921 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7981 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     8041 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     8101 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     8161 tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt tcccatgatt
     8221 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
     8281 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
     8341 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     8401 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgc catgcagtgg
     8461 aatggtaagg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     8521 tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa
     8581 taaggctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct ctagaggtac
     8641 ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     8701 tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     8761 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     8821 acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc ttatgggact
     8881 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
     8941 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
     9001 ttatttttta attattttgt gcagcgatgg gggcggggg
//
