LOCUS       sequence-181565-        9109 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses 3xFLAG-Cas9 and a gRNA targeting mouse Ezh2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9109)
  AUTHORS   Vo BT, Li C, Morgan MA, Theurillat I, Finkelstein D, Wright S, Hyle 
            J, Smith SM, Fan Y, Wang YD, Wu G, Orr BA, Northcott PA, Shilatifard
            A, Sherr CJ, Roussel MF
  TITLE     Inactivation of Ezh2 Upregulates Gfi1 and Drives Aggressive 
            Myc-Driven Group 3 Medulloblastoma.
  JOURNAL   Cell Rep. 2017 Mar 21;18(12):2907-2917. doi: 
            10.1016/j.celrep.2017.02.073.
  PUBMED    28329683
REFERENCE   2  (bases 1 to 9109)
  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.celrep.2017.02"; journalName: "Cell Rep"; date: 
            "2017-03-21- 21"; volume: "18"; issue: "12"; pages: "2907-2917"
COMMENT     Sequence Label: AOI-WT-Cas9-sg-mouse Ezh2-E18-GFP
FEATURES             Location/Qualifiers
     source          1..9109
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      224..233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             233..298
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             305..325
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             350..4450
                     /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             350..4450
                     /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             4451..4498
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4514..4567
                     /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             join(4568..4570,4571..5281)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4568..4570,4571..5281)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4871..4890)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5218..5239
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5309..5326)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5315..5522
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5531..5671
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5531..5660
                     /label=AAV2 ITR
     rep_origin      5746..6201
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5833..5852)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6043..6064
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6218..6237)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6337..6359
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6397..6415)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6483..6587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6588..6656,6657..7448)
                     /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(6806..6825)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7619..8207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8108..8127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8269..8517
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8269..8289
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8440..8459
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8538..8613
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8710..8995
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
       61 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      121 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
      181 tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa
      241 ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg acgataagat
      301 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta
      361 cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
      421 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
      481 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
      541 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
      601 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
      661 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
      721 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
      781 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
      841 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
      901 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
      961 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     1021 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     1081 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     1141 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     1201 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     1261 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     1321 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     1381 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     1441 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     1501 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     1561 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     1621 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     1681 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     1741 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     1801 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     1861 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     1921 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     1981 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     2041 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     2101 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     2161 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     2221 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     2281 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     2341 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     2401 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     2461 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     2521 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     2581 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     2641 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     2701 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     2761 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     2821 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
     2881 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     2941 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     3001 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     3061 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     3121 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     3181 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     3241 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     3301 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     3361 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     3421 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     3481 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     3541 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     3601 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     3661 cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
     3721 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
     3781 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     3841 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     3901 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     3961 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
     4021 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     4081 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
     4141 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     4201 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     4261 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
     4321 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
     4381 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     4441 gggaggcgac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagga
     4501 attcggcagt ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc
     4561 tggcccagtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4621 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     4681 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     4741 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     4801 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4861 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     4921 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     4981 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     5041 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     5101 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     5161 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     5221 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     5281 ggaattctaa ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     5341 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     5401 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     5461 ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg
     5521 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     5581 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     5641 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     5701 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     5761 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5821 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5881 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5941 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     6001 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6061 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     6121 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     6181 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     6241 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     6301 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     6361 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     6421 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6481 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6541 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6601 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6661 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6721 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6781 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6841 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6901 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6961 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7021 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7081 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7141 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7201 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7261 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg
     7321 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7381 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7441 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7501 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7561 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7621 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7681 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7741 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7801 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7861 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7921 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7981 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8041 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8101 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8161 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8221 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtga gggcctattt
     8281 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt
     8341 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
     8401 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
     8461 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgtg
     8521 gtggatgcaa cccgaaagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     8581 tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct agaaatagca
     8641 agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac aaatggctct
     8701 agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     8761 ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc aatgggtgga
     8821 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     8881 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt acatgacctt
     8941 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     9001 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     9061 gtatttattt attttttaat tattttgtgc agcgatgggg gcggggggg
//
