LOCUS       sequence-407539-        9195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cas9 and human H3.2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9195)
  AUTHORS   Kato-Inui T, Takahashi G, Ono T, Miyaoka Y
  TITLE     Fusion of histone variants to Cas9 suppresses non-homologous end 
            joining.
  JOURNAL   PLoS One. 2024 May 13;19(5):e0288578. doi: 
            10.1371/journal.pone.0288578. eCollection 2024.
  PUBMED    38739603
REFERENCE   2  (bases 1 to 9195)
  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: "PLoS One.";
            date: "2024-05-13"; volume: "19(5):e0288578. doi"; pages: " 
            10.1371/journal.pone.0288578. eCollection 2024"
COMMENT     Sequence Label: pCPH3.2-GS3
FEATURES             Location/Qualifiers
     source          1..9195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..297
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        299..576
                     /label=chicken beta-actin promoter
     intron          577..804
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      816..825
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             825..890
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             897..917
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1392..5492
                     /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             1392..5492
                     /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             5493..5540
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5556..5609
                     /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             5610..6209
                     /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(5610..5629)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6106..6126
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6237..6254)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6243..6450
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6459..6599
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6459..6588
                     /label=AAV2 ITR
     rep_origin      6674..7129
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6761..6780)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6971..6992
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7146..7165)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7265..7287
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7325..7343)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7411..7515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7516..7584,7585..8376)
                     /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(7734..7753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8547..9135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9036..9055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
       61 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
      121 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      181 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
      241 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
      301 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
      361 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
      421 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      481 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      541 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      601 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      661 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gctgagcaag
      721 aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta cctggagcac
      781 ctgcctgaaa tcactttttt tcaggttgga ccggtgccac catggactat aaggaccacg
      841 acggagacta caaggatcat gatattgatt acaaagacga tgacgataag atggccccaa
      901 agaagaagcg gaaggtcggg atccacggag tccctgcagc cgcccgaacc aagcagactg
      961 ctcgtaagtc cacaggtggg aaagcccccc gcaaacagct ggccacgaaa gccgccagga
     1021 aaagcgctcc cgctaccggc ggggtgaaga agcctcatcg ctacaggccc gggaccgtgg
     1081 cgcttcgaga gattcgtcgt tatcagaagt cgaccgagct gctcatccgg aagctgccct
     1141 tccagaggtt ggtgagggag atcgcgcagg atttcaaaac cgacctgagg tttcagagct
     1201 cagccgtcat ggcgctgcag gaggctagcg aagcgtacct ggtgggtctg ttcgaagata
     1261 ccaacctgtg tgccatccac gctaagagag tcaccatcat gcccaaagac atccagttgg
     1321 ctcgccggat acggggagag agagctggtg gcggtggatc cggtggcggt ggaagtggtg
     1381 gcggtggatc tgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
     1441 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     1501 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     1561 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     1621 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     1681 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     1741 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     1801 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     1861 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     1921 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     1981 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     2041 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     2101 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     2161 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     2221 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     2281 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     2341 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     2401 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     2461 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     2521 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     2581 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     2641 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     2701 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     2761 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     2821 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     2881 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     2941 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     3001 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     3061 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     3121 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     3181 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     3241 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     3301 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     3361 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     3421 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     3481 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     3541 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     3601 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     3661 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     3721 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     3781 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     3841 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     3901 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     3961 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     4021 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     4081 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     4141 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     4201 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     4261 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     4321 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     4381 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     4441 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     4501 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     4561 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     4621 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     4681 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     4741 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     4801 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     4861 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     4921 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     4981 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     5041 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     5101 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     5161 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     5221 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     5281 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     5341 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     5401 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     5461 cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg
     5521 gccaggcaaa aaagaaaaag gaattcggca gtggagaggg cagaggaagt ctgctaacat
     5581 gcggtgacgt cgaggagaat cctggcccaa tgaccgagta caagcccacg gtgcgcctcg
     5641 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     5701 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     5761 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     5821 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     5881 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     5941 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct
     6001 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     6061 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     6121 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     6181 ggtgcatgac ccgcaagccc ggtgcctgag aattctaact agagctcgct gatcagcctc
     6241 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     6301 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     6361 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     6421 ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt
     6481 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     6541 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     6601 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     6661 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     6721 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     6781 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6841 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     6901 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6961 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     7021 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     7081 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     7141 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     7201 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     7261 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     7321 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     7381 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     7441 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     7501 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     7561 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     7621 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     7681 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     7741 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     7801 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7861 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7921 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7981 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     8041 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     8101 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     8161 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     8221 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     8281 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     8341 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     8401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     8461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     8521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     8581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     8641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     8701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     8761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     8821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     9001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     9061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     9121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     9181 ccttttgctc acatg
//
