LOCUS       sequence-430623-       10664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to 
            purify specific genomic regions of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10664)
  AUTHORS   Yuno M, Nagata S, Fujita T, Fujii H
  TITLE     MSCV-based retroviral plasmids expressing 3xFLAG-Sp-dCas9 for enChIP
            analysis.
  JOURNAL   Biol Methods Protoc. 2021 Jul 9;6(1):bpab013. doi: 
            10.1093/biomethods/bpab013. eCollection 2021.
  PUBMED    34409168
REFERENCE   2  (bases 1 to 10664)
  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: "Biol 
            Methods Protoc."; date: "2021-07-9"; volume: "6(1):bpab013. doi"; 
            pages: " 10.1093/biomethods/bpab013. eCollection 2021"
COMMENT     Sequence Label: 3xFLAG-dCas9/pMSCVpuro
FEATURES             Location/Qualifiers
     source          1..10664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1452..1517
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(1521..1547,1548..1550,1551..4034,4035..4040,4041..4106,
                     4107..4109,4110..5624)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1521..1547,1548..1550,1551..4037,4038..4040,
                     4041..5624)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1521..5624
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5637..5657
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        5838..6337
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5873..5896)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6270..6289
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6358..6957
                     /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(6358..6377)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6854..6874
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7057..7571
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7733..7749
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7757..7763,7764..7781,7782..7787))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7802..7823
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7940..7957)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8111..8699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8191..8210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8870..9661,9662..9730))
                     /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     9493..9512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9731..9835)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9903..9921
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9959..9981)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10081..10100
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10294..10316
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10444..10463
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(10663..10664,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctaagga tccgtttagt gaaccgtcag
     1441 aattaaccat ggactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca
     1501 aggatgacga tgacaagctc atggacaaga agtactccat tgggctcgct atcggcacaa
     1561 acagcgtcgg ctgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag
     1621 ttctgggcaa taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg
     1681 actccgggga gacggccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc
     1741 gcagaaagaa tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg
     1801 atgactcttt cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagcacg
     1861 agcgccaccc aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa
     1921 ccatatatca tctgaggaag aagcttgtag acagtactga taaggctgac ttgcggttga
     1981 tctatctcgc gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc
     2041 tgaacccaga caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc
     2101 agcttttcga agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg
     2161 ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga
     2221 agaacggcct gtttggtaat cttatcgccc tgtcactcgg gctgaccccc aactttaaat
     2281 ctaacttcga cctggccgaa gatgccaagc ttcaactgag caaagacacc tacgatgatg
     2341 atctcgacaa tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa
     2401 agaacctgtc agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca
     2461 aagctccgct gagcgctagt atgatcaagc gctatgatga gcaccaccaa gacttgactt
     2521 tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc
     2581 agtctaaaaa tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca
     2641 aatttattaa gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta
     2701 acagagaaga tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc ccccaccaga
     2761 ttcacctggg cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga
     2821 aagataacag ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc
     2881 ccctcgcccg gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca
     2941 ctccctggaa cttcgaggaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa
     3001 ggatgactaa ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc
     3061 tgtacgagta cttcacagtt tataacgagc tcaccaaggt caaatacgtc acagaaggga
     3121 tgagaaagcc agcattcctg tctggagagc agaagaaagc tatcgtggac ctcctcttca
     3181 agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat
     3241 gtttcgactc tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt
     3301 atcacgatct cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg
     3361 acattcttga ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag
     3421 aacgcttgaa aacttacgct catctcttcg acgacaaagt catgaaacag ctcaagaggc
     3481 gccgatatac aggatggggg cggctgtcaa gaaaactgat caatgggatc cgagacaagc
     3541 agagtggaaa gacaatcctg gattttctta agtccgatgg atttgccaac aggaacttca
     3601 tgcagttgat ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt
     3661 ctggccaggg ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca
     3721 aaaagggaat actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc
     3781 ataagcccga gaatatcgtt atcgagatgg cccgagagaa ccaaactacc cagaagggac
     3841 agaagaacag tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc
     3901 aaatccttaa ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt
     3961 actacctgca gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct
     4021 ccgactacga cgtggatgcc atcgtgcccc agtcttttct caaagatgat tctattgata
     4081 ataaagtgtt gacaagatcc gataaaaata gagggaagag tgataacgtc ccctcagaag
     4141 aagttgtcaa gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac
     4201 aacggaagtt cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag
     4261 ccggcttcat caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa
     4321 ttctcgattc acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga
     4381 aagttattac tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata
     4441 aggtgagaga gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag
     4501 gcactgcact tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata
     4561 aagtgtacga tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg
     4621 ctaagtactt cttttacagc aatattatga attttttcaa gaccgagatt acactggcca
     4681 atggagagat tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt
     4741 gggacaaggg tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca
     4801 tcgttaaaaa gaccgaagta cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa
     4861 ggaacagcga caagctgatc gcacgcaaaa aagattggga ccccaagaaa tacggcggat
     4921 tcgattctcc tacagtcgct tacagtgtac tggttgtggc caaagtggag aaagggaagt
     4981 ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac aatcatggag cgatcaagct
     5041 tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata taaagaggtc aaaaaagacc
     5101 tcatcattaa gcttcccaag tactctctct ttgagcttga aaacggccgg aaacgaatgc
     5161 tcgctagtgc gggcgagctg cagaaaggta acgagctggc actgccctct aaatacgtta
     5221 atttcttgta tctggccagc cactatgaaa agctcaaagg gtctcccgaa gataatgagc
     5281 agaagcagct gttcgtggaa caacacaaac actaccttga tgagatcatc gagcaaataa
     5341 gcgaattctc caaaagagtg atcctcgccg acgctaacct cgataaggtg ctttctgctt
     5401 acaataagca cagggataag cccatcaggg agcaggcaga aaacattatc cacttgttta
     5461 ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt cgacaccacc atagacagaa
     5521 agcggtacac ctctacaaag gaggtcctgg acgccacact gattcatcag tcaattacgg
     5581 ggctctatga aacaagaatc gacctctctc agctcggtgg agacagcagg gctgacccca
     5641 agaagaagag gaaggtgtga aagggttcga tccctaccgg atcgaattcc agctgagcgc
     5701 cggtcgctac cattaccagt tggtctggtg tcaaaaataa taataaccgg gcaggccatg
     5761 tctgcccgta tttcgcgtaa ggaaatccat tatgtactat ttaaactcga gcggcctcga
     5821 ggttaacgaa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
     5881 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
     5941 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
     6001 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
     6061 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
     6121 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
     6181 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
     6241 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
     6301 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg
     6361 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     6421 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     6481 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     6541 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     6601 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     6661 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     6721 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     6781 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     6841 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     6901 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc
     6961 ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgataaaa
     7021 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     7081 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
     7141 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
     7201 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     7261 ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     7321 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     7381 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
     7441 gatagactgc gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     7501 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     7561 gggtctttca tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc
     7621 gaatattaag taatcctgac tcaattagcc actgttttga atccacatac tccaatactc
     7681 ctgaaatagt tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc
     7741 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     7801 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     8041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     8101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     8161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     8281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     8341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     8401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     8581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8641 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8701 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     8761 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8821 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     9001 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     9061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     9121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     9181 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     9241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     9301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     9361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     9421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     9481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9721 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9841 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     9901 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     9961 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
    10021 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
    10081 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
    10141 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
    10201 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
    10261 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
    10321 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
    10381 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
    10441 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
    10501 cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt
    10561 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata
    10621 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaa
//
