LOCUS       sequence-171514-        9011 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses sgRNA targeting human TP53 around codon F109.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9011)
  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 9011)
  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: sgTP53(B)
FEATURES             Location/Qualifiers
     source          1..9011
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      238..247
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             247..312
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             319..339
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             364..4464
                     /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             364..4464
                     /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             4465..4512
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4528..4581
                     /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             4582..5181
                     /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(4582..4601)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(5209..5226)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5215..5422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5431..5571
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5431..5560
                     /label=AAV2 ITR
     rep_origin      5646..6101
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5733..5752)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5943..5964
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6118..6137)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6237..6259
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6297..6315)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6383..6487
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6488..6556,6557..7348)
                     /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(6706..6725)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7519..8107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8008..8027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8169..8417
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8169..8189
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8340..8359
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8439..8514
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8611..8896
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
       61 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      121 gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt
      181 aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcc
      241 accatggact ataaggacca cgacggagac tacaaggatc atgatattga ttacaaagac
      301 gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca
      361 gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg
      421 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
      481 cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag
      541 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
      601 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
      661 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
      721 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
      781 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
      841 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
      901 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
      961 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     1021 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac
     1081 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     1141 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     1201 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     1261 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     1321 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     1381 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     1441 ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     1501 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     1561 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     1621 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     1681 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     1741 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     1801 gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     1861 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     1921 tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     1981 agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     2041 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     2101 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     2161 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     2221 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     2281 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     2341 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     2401 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     2461 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     2521 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     2581 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     2641 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     2701 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     2761 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     2821 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat
     2881 atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc
     2941 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     3001 aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg
     3061 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     3121 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     3181 actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc
     3241 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     3301 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     3361 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     3421 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     3481 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     3541 cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt
     3601 gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     3661 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc
     3721 gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc
     3781 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
     3841 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
     3901 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
     3961 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg
     4021 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
     4081 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
     4141 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
     4201 atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag
     4261 cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
     4321 cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa
     4381 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
     4441 gacctgtctc agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca
     4501 aaaaagaaaa aggaattcgg cagtggagag ggcagaggaa gtctgctaac atgcggtgac
     4561 gtcgaggaga atcctggccc aatgaccgag tacaagccca cggtgcgcct cgccacccgc
     4621 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4681 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4741 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4801 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4861 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4921 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4981 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5041 gccggggtgc ccgccttcct ggagacctcc gcgccccaca acctcccctt ctacgagcgg
     5101 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5161 acccgcaagc ccggtgcctg agaattctaa ctagagctcg ctgatcagcc tcgactgtgc
     5221 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     5281 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     5341 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     5401 agaatagcag gcatgctggg gagcggccgc aggaacccct agtgatggag ttggccactc
     5461 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     5521 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     5581 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     5641 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5701 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     5761 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5821 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5881 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5941 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     6001 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     6061 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     6121 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     6181 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     6241 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     6301 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     6361 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6421 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6481 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6541 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6601 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6661 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6721 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6781 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6841 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6901 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6961 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7021 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7081 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7141 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7201 tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7261 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7321 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7381 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7441 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7501 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7561 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7621 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7681 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7741 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7801 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7861 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7921 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7981 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8041 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8101 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8161 tcacatgtga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     8221 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     8281 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     8341 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
     8401 ggaaaggacg aaacaccgca gaatgcaaga agcccagagt tttagagcta gaaatagcaa
     8461 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     8521 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgtttttagc gcgtgcgcca
     8581 attctgcaga caaatggctc tagaggtacc cgttacataa cttacggtaa atggcccgcc
     8641 tggctgaccg cccaacgacc cccgcccatt gacgtcaata gtaacgccaa tagggacttt
     8701 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     8761 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     8821 ttgtgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     8881 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     8941 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     9001 ggcggggggg g
//
