LOCUS       sequence-430627-       10842 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 10842)
  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 10842)
  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/pMSCVneo
FEATURES             Location/Qualifiers
     source          1..10842
                     /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             1465..1530
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(1534..1560,1561..1563,1564..4047,4048..4053,4054..4119,
                     4120..4122,4123..5637)
                     /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(1534..1560,1561..1563,1564..4050,4051..4053,
                     4054..5637)
                     /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             1534..5637
                     /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             5650..5670
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        5844..6343
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5879..5902)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6276..6295
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6357..7160
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6420..6439)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7030..7049
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             7235..7749
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7911..7927
                     /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(7935..7941,7942..7959,7960..7965))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7980..8001
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8118..8135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8289..8877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8369..8388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9048..9839,9840..9908))
                     /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     9671..9690
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9909..10013)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10081..10099
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(10137..10159)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10259..10278
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10472..10494
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10622..10641
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(10841..10842,1..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 gccggaattc gttaacctcg agagatctaa ggatccgttt
     1441 agtgaaccgt cagaattaac catggactac aaagaccatg acggtgatta taaagatcat
     1501 gacatcgatt acaaggatga cgatgacaag ctcatggaca agaagtactc cattgggctc
     1561 gctatcggca caaacagcgt cggctgggcc gtcattacgg acgagtacaa ggtgccgagc
     1621 aaaaaattca aagttctggg caataccgat cgccacagca taaagaagaa cctcattggc
     1681 gccctcctgt tcgactccgg ggagacggcc gaagccacgc ggctcaaaag aacagcacgg
     1741 cgcagatata cccgcagaaa gaatcggatc tgctacctgc aggagatctt tagtaatgag
     1801 atggctaagg tggatgactc tttcttccat aggctggagg agtccttttt ggtggaggag
     1861 gataaaaagc acgagcgcca cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat
     1921 gaaaagtacc caaccatata tcatctgagg aagaagcttg tagacagtac tgataaggct
     1981 gacttgcggt tgatctatct cgcgctggcg catatgatca aatttcgggg acacttcctc
     2041 atcgaggggg acctgaaccc agacaacagc gatgtcgaca aactctttat ccaactggtt
     2101 cagacttaca atcagctttt cgaagagaac ccgatcaacg catccggagt tgacgccaaa
     2161 gcaatcctga gcgctaggct gtccaaatcc cggcggctcg aaaacctcat cgcacagctc
     2221 cctggggaga agaagaacgg cctgtttggt aatcttatcg ccctgtcact cgggctgacc
     2281 cccaacttta aatctaactt cgacctggcc gaagatgcca agcttcaact gagcaaagac
     2341 acctacgatg atgatctcga caatctgctg gcccagatcg gcgaccagta cgcagacctt
     2401 tttttggcgg caaagaacct gtcagacgcc attctgctga gtgatattct gcgagtgaac
     2461 acggagatca ccaaagctcc gctgagcgct agtatgatca agcgctatga tgagcaccac
     2521 caagacttga ctttgctgaa ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa
     2581 attttcttcg atcagtctaa aaatggctac gccggataca ttgacggcgg agcaagccag
     2641 gaggaatttt acaaatttat taagcccatc ttggaaaaaa tggacggcac cgaggagctg
     2701 ctggtaaagc ttaacagaga agatctgttg cgcaaacagc gcactttcga caatggaagc
     2761 atcccccacc agattcacct gggcgaactg cacgctatcc tcaggcggca agaggatttc
     2821 tacccctttt tgaaagataa cagggaaaag attgagaaaa tcctcacatt tcggataccc
     2881 tactatgtag gccccctcgc ccggggaaat tccagattcg cgtggatgac tcgcaaatca
     2941 gaagagacca tcactccctg gaacttcgag gaagtcgtgg ataagggggc ctctgcccag
     3001 tccttcatcg aaaggatgac taactttgat aaaaatctgc ctaacgaaaa ggtgcttcct
     3061 aaacactctc tgctgtacga gtacttcaca gtttataacg agctcaccaa ggtcaaatac
     3121 gtcacagaag ggatgagaaa gccagcattc ctgtctggag agcagaagaa agctatcgtg
     3181 gacctcctct tcaagacgaa ccggaaagtt accgtgaaac agctcaaaga agactatttc
     3241 aaaaagattg aatgtttcga ctctgttgaa atcagcggag tggaggatcg cttcaacgca
     3301 tccctgggaa cgtatcacga tctcctgaaa atcattaaag acaaggactt cctggacaat
     3361 gaggagaacg aggacattct tgaggacatt gtcctcaccc ttacgttgtt tgaagatagg
     3421 gagatgattg aagaacgctt gaaaacttac gctcatctct tcgacgacaa agtcatgaaa
     3481 cagctcaaga ggcgccgata tacaggatgg gggcggctgt caagaaaact gatcaatggg
     3541 atccgagaca agcagagtgg aaagacaatc ctggattttc ttaagtccga tggatttgcc
     3601 aacaggaact tcatgcagtt gatccatgat gactctctca cctttaagga ggacatccag
     3661 aaagcacaag tttctggcca gggggacagt cttcacgagc acatcgctaa tcttgcaggt
     3721 agcccagcta tcaaaaaggg aatactgcag accgttaagg tcgtggatga actcgtcaaa
     3781 gtaatgggaa ggcataagcc cgagaatatc gttatcgaga tggcccgaga gaaccaaact
     3841 acccagaagg gacagaagaa cagtagggaa aggatgaaga ggattgaaga gggtataaaa
     3901 gaactggggt cccaaatcct taaggaacac ccagttgaaa acacccagct tcagaatgag
     3961 aagctctacc tgtactacct gcagaacggc agggacatgt acgtggatca ggaactggac
     4021 atcaatcggc tctccgacta cgacgtggat gccatcgtgc cccagtcttt tctcaaagat
     4081 gattctattg ataataaagt gttgacaaga tccgataaaa atagagggaa gagtgataac
     4141 gtcccctcag aagaagttgt caagaaaatg aaaaattatt ggcggcagct gctgaacgcc
     4201 aaactgatca cacaacggaa gttcgataat ctgactaagg ctgaacgagg tggcctgtct
     4261 gagttggata aagccggctt catcaaaagg cagcttgttg agacacgcca gatcaccaag
     4321 cacgtggccc aaattctcga ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg
     4381 attcgagagg tgaaagttat tactctgaag tctaagctgg tctcagattt cagaaaggac
     4441 tttcagtttt ataaggtgag agagatcaac aattaccacc atgcgcatga tgcctacctg
     4501 aatgcagtgg taggcactgc acttatcaaa aaatatccca agcttgaatc tgaatttgtt
     4561 tacggagact ataaagtgta cgatgttagg aaaatgatcg caaagtctga gcaggaaata
     4621 ggcaaggcca ccgctaagta cttcttttac agcaatatta tgaatttttt caagaccgag
     4681 attacactgg ccaatggaga gattcggaag cgaccactta tcgaaacaaa cggagaaaca
     4741 ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag tccggaaggt cctgtccatg
     4801 ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg gaggcttctc caaggaaagt
     4861 atcctcccga aaaggaacag cgacaagctg atcgcacgca aaaaagattg ggaccccaag
     4921 aaatacggcg gattcgattc tcctacagtc gcttacagtg tactggttgt ggccaaagtg
     4981 gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac tgctgggcat cacaatcatg
     5041 gagcgatcaa gcttcgaaaa aaaccccatc gactttctcg aggcgaaagg atataaagag
     5101 gtcaaaaaag acctcatcat taagcttccc aagtactctc tctttgagct tgaaaacggc
     5161 cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc
     5221 tctaaatacg ttaatttctt gtatctggcc agccactatg aaaagctcaa agggtctccc
     5281 gaagataatg agcagaagca gctgttcgtg gaacaacaca aacactacct tgatgagatc
     5341 atcgagcaaa taagcgaatt ctccaaaaga gtgatcctcg ccgacgctaa cctcgataag
     5401 gtgctttctg cttacaataa gcacagggat aagcccatca gggagcaggc agaaaacatt
     5461 atccacttgt ttactctgac caacttgggc gcgcctgcag ccttcaagta cttcgacacc
     5521 accatagaca gaaagcggta cacctctaca aaggaggtcc tggacgccac actgattcat
     5581 cagtcaatta cggggctcta tgaaacaaga atcgacctct ctcagctcgg tggagacagc
     5641 agggctgacc ccaagaagaa gaggaaggtg tgaaagggtt cgatccctac cggatcgaat
     5701 tccagctgag cgccggtcgc taccattacc agttggtctg gtgtcaaaaa taataataac
     5761 cgggcaggcc atgtctgccc gtatttcgcg taaggaaatc cattatgtac tatttaaact
     5821 cgagcggccg atctaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc
     5881 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac
     5941 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc
     6001 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
     6061 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
     6121 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
     6181 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
     6241 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
     6301 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gccaatatgg
     6361 gatcggccat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     6421 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     6481 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6541 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6601 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6661 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6721 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6781 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6841 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     6901 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6961 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     7021 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     7081 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     7141 ttcttgacga gttcttctga ggggatccgt cgacctgcag ccaagcttat cgataaaata
     7201 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     7261 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     7321 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     7381 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     7441 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     7501 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     7561 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     7621 tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     7681 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     7741 gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga
     7801 atattaagta atcctgactc aattagccac tgttttgaat ccacatactc caatactcct
     7861 gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg
     7921 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     7981 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     8041 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     8101 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     8161 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     8221 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     8281 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     8341 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     8401 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     8461 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     8521 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     8581 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     8641 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     8701 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     8761 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     8821 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     8881 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     8941 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     9001 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     9061 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     9121 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     9181 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     9241 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     9301 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     9361 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     9421 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     9481 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     9541 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     9601 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     9661 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     9721 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     9781 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     9841 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     9901 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     9961 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
    10021 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
    10081 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
    10141 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
    10201 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
    10261 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
    10321 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
    10381 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
    10441 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg
    10501 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
    10561 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
    10621 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
    10681 cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc
    10741 aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga
    10801 taaaataaaa gattttattt agtctccaga aaaagggggg aa
//
