LOCUS       sequence-97110-e        9442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses D10A mutant (nickase) of human codon-optimized Cas9 
            (derived from Streptococcus pyogenes) and pruomycin resistance 
            gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9442)
  AUTHORS   Li HL, Fujimoto N, Sasakawa N, Shirai S, Ohkame T, Sakuma T, Tanaka 
            M, Amano N, Watanabe A, Sakurai H, Yamamoto T, Yamanaka S, Hotta A
  TITLE     Precise Correction of the Dystrophin Gene in Duchenne Muscular 
            Dystrophy Patient Induced Pluripotent Stem Cells by TALEN and 
            CRISPR-Cas9.
  JOURNAL   Stem Cell Reports. 2014 Nov 25. pii: S2213-6711(14)00335-X. doi: 
            10.1016/j.stemcr.2014.10.013.
  PUBMED    25434822
REFERENCE   2  (bases 1 to 9442)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cell 
            Reports. 2014 Nov 25. pii: S2213-6711(14)00335-X. doi: 
            10.1016/j.stemcr.2014.10.013."
COMMENT     Sequence Label: pHL-EF1a-SphcCas9(D10A)-iP-A
FEATURES             Location/Qualifiers
     source          1..9442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        157..1339
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          388..1330
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1287..1307
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1361..1385
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1411..1437,1438..1440,1441..5514)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     RBS             5427..5435
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             5527..5547
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(5560..5584)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(5626..6200,6201..6203,6204..6212)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5793..5810)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6020..6039
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6213..6812
                     /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(6213..6232)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6709..6729
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7000..7019)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7065..7120
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7119..7138)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7536..7553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7707..8295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7787..7806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8466..9257,9258..9326))
                     /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     9089..9108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9327..9431)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaaaagtgcc acctgacgtc gctagcgttt aaacccgggc gatcgctcta gactagtgcg
       61 gccgctgcag aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc
      121 taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca
      181 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
      241 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
      301 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
      361 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
      421 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacgcccct ggctgcagta
      481 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
      541 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
      601 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
      661 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
      721 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
      781 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
      841 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
      901 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
      961 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     1021 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     1081 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     1141 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1201 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1261 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     1321 tccatttcag gtgtcgtgag aattagcggt ggcggccatc acaagtttgt acaaaaaagc
     1381 aggctggcgc cggaaccaat tcagtcgacc atggataaga aatacagcat tggactggcc
     1441 attgggacaa actccgtggg atgggccgtg attacagacg aatacaaagt gccttcaaag
     1501 aagttcaagg tgctgggcaa caccgataga cacagcatca agaaaaatct gattggagcc
     1561 ctgctgttcg actccggcga gacagctgaa gcaactcggc tgaaaagaac tgctcggaga
     1621 aggtataccc gccgaaagaa taggatctgc tacctgcagg agattttcag caacgaaatg
     1681 gccaaggtgg acgatagttt ctttcaccgc ctggaggaat cattcctggt cgaggaagat
     1741 aagaaacacg agcggcatcc catctttggc aacattgtgg acgaggtcgc ttatcacgaa
     1801 aagtacccta ccatctatca tctgaggaag aaactggtgg actccacaga taaagcagac
     1861 ctgcgcctga tctatctggc cctggctcac atgattaagt tccggggcca ttttctgatc
     1921 gagggggatc tgaacccaga caattctgat gtggacaagc tgttcatcca gctggtccag
     1981 acatacaatc agctgtttga ggaaaacccc attaatgcat ctggcgtgga cgcaaaagcc
     2041 atcctgagtg ccagactgtc taagagtcgg agactggaga acctgatcgc tcagctgcca
     2101 ggggaaaaga aaaacggcct gtttgggaat ctgattgcac tgtcactggg actgactccc
     2161 aacttcaaga gcaattttga tctggccgag gacgctaaac tgcagctgtc caaggacacc
     2221 tatgacgatg acctggataa cctgctggct cagatcgggg atcagtacgc agacctgttc
     2281 ctggccgcta agaatctgtc tgacgccatc ctgctgagtg atattctgcg cgtgaacacc
     2341 gagattacaa aagcccccct gtcagctagc atgatcaaga gatatgacga gcaccatcag
     2401 gatctgaccc tgctgaaggc tctggtgagg cagcagctgc ctgagaagta caaggaaatc
     2461 ttctttgatc agtctaagaa cggatacgcc ggctatattg acggcggggc tagtcaggag
     2521 gagttctaca agtttatcaa acccattctg gagaagatgg atggcacaga ggaactgctg
     2581 gtgaaactga atcgggaaga cctgctgagg aagcagcgca cttttgataa cggaagcatc
     2641 cctcaccaga ttcatctggg agagctgcac gcaatcctga ggcgccagga agacttctac
     2701 ccatttctga aggataacag ggagaagatc gaaaaaattc tgacattccg catcccctac
     2761 tatgtgggcc ctctggcaag aggcaacagc cggtttgcct ggatgactcg caaatctgag
     2821 gaaacaatca ctccctggaa cttcgaggaa gtggtcgata agggcgcttc cgcacagtct
     2881 ttcattgagc ggatgacaaa cttcgacaag aacctgccaa acgaaaaagt gctgcccaag
     2941 cactctctgc tgtacgagta tttcacagtc tataacgaac tgactaaggt gaaatacgtc
     3001 accgagggga tgagaaagcc tgccttcctg agtggagaac agaagaaagc tatcgtggac
     3061 ctgctgttta aaaccaatag gaaggtgaca gtcaagcagc tgaaagagga ctatttcaag
     3121 aaaattgaat gtttcgattc tgtggagatc agtggcgtcg aagacaggtt taacgcctcc
     3181 ctggggacct accacgatct gctgaagatc attaaggata aagacttcct ggacaacgag
     3241 gaaaatgagg atatcctgga agacattgtg ctgaccctga cactgtttga ggatagggaa
     3301 atgatcgagg aacgcctgaa gacctatgcc catctgttcg atgacaaagt gatgaaacag
     3361 ctgaagcgac ggagatacac aggatggggc cgactgtctc ggaagctgat caatgggatt
     3421 cgcgacaaac agagtggaaa gaccatcctg gactttctga aatcagatgg cttcgccaac
     3481 cggaacttca tgcagctgat tcacgatgac agcctgacat tcaaagagga tatccagaag
     3541 gcacaggtgt ccgggcaggg agactctctg cacgagcata tcgcaaacct ggccggcagc
     3601 cctgccatca agaaagggat tctgcagacc gtgaaggtgg tggacgagct ggtgaaagtc
     3661 atgggaagac ataagccaga aaacatcgtg attgagatgg ccagggaaaa tcagaccaca
     3721 cagaaaggcc agaagaactc aagggagcgc atgaaaagaa tcgaggaagg aattaaggaa
     3781 ctgggcagcc agatcctgaa agagcacccc gtggaaaaca cacagctgca gaatgagaag
     3841 ctgtatctgt actatctgca gaatggacgc gatatgtacg tggaccagga gctggatatt
     3901 aaccgactgt ccgattacga cgtggatcat atcgtcccac agtcattcct gaaagatgac
     3961 agcattgaca ataaggtgct gacccgctct gacaaaaacc gaggcaagag tgataatgtc
     4021 ccctcagagg aagtggtcaa gaaaatgaag aactactgga ggcagctgct gaatgccaaa
     4081 ctgatcacac agcgaaagtt tgataacctg actaaagctg agcggggagg cctgagtgaa
     4141 ctggacaaag caggcttcat taagcgacag ctggtggaga cacggcagat cacaaagcac
     4201 gtcgcccaga ttctggattc aagaatgaac actaagtacg atgagaatga caaactgatc
     4261 agagaagtga aggtcattac cctgaagtca aaactggtga gcgactttcg gaaagatttc
     4321 cagttttata aggtcagaga gatcaacaac taccaccatg ctcatgacgc atacctgaac
     4381 gcagtggtcg gcacagccct gattaagaaa taccctaaac tggagtccga gttcgtgtac
     4441 ggggactata aggtgtacga tgtcagaaaa atgatcgcca agtctgagca ggaaattggc
     4501 aaagccactg ctaagtattt cttttacagt aacatcatga atttctttaa gactgagatc
     4561 accctggcaa atggggaaat ccgaaagcgg ccactgattg agactaacgg cgagacagga
     4621 gaaatcgtgt gggacaaagg aagagatttt gctaccgtga ggaaggtcct gagcatgccc
     4681 caagtgaata ttgtcaagaa aacagaggtg cagactgggg gattcagtaa ggaatcaatt
     4741 ctgcctaaac gcaactccga taagctgatc gcccgaaaga aagactggga ccccaagaag
     4801 tatggcgggt tcgactcccc aactgtggct tactctgtcc tggtggtcgc aaaggtggag
     4861 aagggaaaaa gcaagaaact gaaatccgtc aaggaactgc tgggcatcac cattatggag
     4921 cgcagctcct tcgaaaagaa tcctatcgat tttctggagg ccaaaggcta taaggaagtg
     4981 aagaaagacc tgatcatcaa gctgccaaag tactcactgt ttgagctgga aaacgggaga
     5041 aagaggatgc tggcaagcgc cggggagctg cagaaaggaa atgaactggc cctgccctcc
     5101 aagtacgtga acttcctgta tctggctagc cactacgaga agctgaaagg gtcccctgag
     5161 gataacgaac agaaacagct gtttgtggag cagcacaagc attatctgga cgagatcatt
     5221 gaacagatta gcgagttctc caaaagagtg atcctggctg acgcaaatct ggataaggtc
     5281 ctgagcgcat acaacaaaca ccgggataag ccaatcagag agcaggccga aaatatcatt
     5341 catctgttca ctctgaccaa cctgggagcc cccgcagcct tcaagtattt tgacactacc
     5401 atcgatcgca aacgatacac aagcactaag gaggtgctgg acgctaccct gattcatcag
     5461 agcattactg gcctgtatga aacaaggatt gacctgtctc agctgggcgg cgactccgga
     5521 gctgacccca agaagaagag gaaggtgtga gacctctaga cccagctttc ttgtacaaag
     5581 tggtgatcta gctaaaacgc ggcctcgaat cgaattccgc ccccccccct ctccctcccc
     5641 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     5701 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     5761 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     5821 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     5881 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     5941 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     6001 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     6061 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     6121 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     6181 ttgaaaaaca cgatgataat atggccacaa ccatgaccga gtacaagccc acggtgcgcc
     6241 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     6301 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     6361 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     6421 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     6481 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     6541 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     6601 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     6661 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     6721 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     6781 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     6841 ccgaaaggag cgcacgaccc catgcatcga ttcgaatgta cagatctgga tcctaggctt
     6901 aagtatacct cgagttaaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc
     6961 tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa
     7021 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     7081 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     7141 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     7201 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     7261 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     7321 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     7381 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     7441 gagatccctc gacttaatta aaccggtcag ctgcattaat gaatcggcca acgcgcgggg
     7501 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7561 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7621 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8401 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8461 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8521 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8641 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8761 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8881 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     9001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9061 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     9121 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9301 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9361 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9421 ggggttccgc gcacatttcc cc
//
