LOCUS       sequence-77870-e       10698 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 10698)
  AUTHORS   Fujita T, Fujii H
  TITLE     Identification of Proteins Associated with an IFNgamma-Responsive 
            Promoter by a Retroviral Expression System for enChIP Using CRISPR.
  JOURNAL   PLoS One. 2014 Jul 22;9(7):e103084. doi: 
            10.1371/journal.pone.0103084. eCollection 2014.
  PUBMED    25051498
REFERENCE   2  (bases 1 to 10698)
  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: "2014-07-22"; volume: "9(7):e103084. doi"; pages: " 
            10.1371/journal.pone.0103084. eCollection 2014"
COMMENT     Sequence Label: 3xFLAG-dCas9/pMXs-I2
FEATURES             Location/Qualifiers
     source          1..10698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1951..2016
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(2020..2046,2047..2049,2050..4533,4534..4539,4540..4605,
                     4606..4608,4609..6123)
                     /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(2020..2046,2047..2049,2050..4536,4537..4539,
                     4540..6123)
                     /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             2020..6123
                     /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             6136..6156
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    6331..6904
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6354..6906
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6498..6515)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6725..6744
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             8203..8672
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(8802..8819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8973..9561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9053..9072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9725..10516,10517..10585))
                     /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     10348..10367
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10586..10690)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1921 ccgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
     1981 gatcatgaca tcgattacaa ggatgacgat gacaagctca tggacaagaa gtactccatt
     2041 gggctcgcta tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg
     2101 ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc
     2161 attggcgccc tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca
     2221 gcacggcgca gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt
     2281 aatgagatgg ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg
     2341 gaggaggata aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg
     2401 taccatgaaa agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat
     2461 aaggctgact tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac
     2521 ttcctcatcg agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa
     2581 ctggttcaga cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac
     2641 gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca
     2701 cagctccctg gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg
     2761 ctgaccccca actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc
     2821 aaagacacct acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca
     2881 gacctttttt tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga
     2941 gtgaacacgg agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag
     3001 caccaccaag acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac
     3061 aaggaaattt tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca
     3121 agccaggagg aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag
     3181 gagctgctgg taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat
     3241 ggaagcatcc cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag
     3301 gatttctacc cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg
     3361 ataccctact atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc
     3421 aaatcagaag agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct
     3481 gcccagtcct tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg
     3541 cttcctaaac actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc
     3601 aaatacgtca cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct
     3661 atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac
     3721 tatttcaaaa agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc
     3781 aacgcatccc tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg
     3841 gacaatgagg agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa
     3901 gatagggaga tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc
     3961 atgaaacagc tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc
     4021 aatgggatcc gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga
     4081 tttgccaaca ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac
     4141 atccagaaag cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt
     4201 gcaggtagcc cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc
     4261 gtcaaagtaa tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac
     4321 caaactaccc agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt
     4381 ataaaagaac tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag
     4441 aatgagaagc tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa
     4501 ctggacatca atcggctctc cgactacgac gtggatgcca tcgtgcccca gtcttttctc
     4561 aaagatgatt ctattgataa taaagtgttg acaagatccg ataaaaatag agggaagagt
     4621 gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg
     4681 aacgccaaac tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc
     4741 ctgtctgagt tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc
     4801 accaagcacg tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac
     4861 aaactgattc gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga
     4921 aaggactttc agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc
     4981 tacctgaatg cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa
     5041 tttgtttacg gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag
     5101 gaaataggca aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag
     5161 accgagatta cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga
     5221 gaaacaggag aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg
     5281 tccatgccgc aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag
     5341 gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac
     5401 cccaagaaat acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc
     5461 aaagtggaga aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca
     5521 atcatggagc gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat
     5581 aaagaggtca aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa
     5641 aacggccgga aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca
     5701 ctgccctcta aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg
     5761 tctcccgaag ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat
     5821 gagatcatcg agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc
     5881 gataaggtgc tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa
     5941 aacattatcc acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc
     6001 gacaccacca tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg
     6061 attcatcagt caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga
     6121 gacagcaggg ctgaccccaa gaagaagagg aaggtgtgaa agggttggat ccctaccgga
     6181 tcgaattcca gctgagcgcc ggtcgctacc attaccagtt ggtctggtgt caaaaataat
     6241 aataaccggg caggccatgt ctgcccgtat ttcgcgtaag gaaatccatt atgtactatt
     6301 taaactcgag cggccgctac gtaaattccg cccctctccc tccccccccc ctaacgttac
     6361 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     6421 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     6481 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     6541 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     6601 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     6661 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     6721 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     6781 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     6841 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     6901 ataatatggc cacaattcct aagatgagct ttccatgtaa atttgtagcc agcttccttc
     6961 tgattttcaa tgtttcttcc aaaggtgcag tctccaaaga gattacgaat gccttggaaa
     7021 cctggggtgc cttgggtcag gacatcaact tggacattcc tagttttcaa atgagtgatg
     7081 atattgacga tataaaatgg gaaaaaactt cagacaagaa aaagattgca caattcagaa
     7141 aagagaaaga gactttcaag gaaaaagata catataagct atttaaaaat ggaactctga
     7201 aaattaagca tctgaagacc gatgatcagg atatctacaa ggtatcaata tatgatacaa
     7261 aaggaaaaaa tgtgttggaa aaaatatttg atttgaagat tcaagagagg gtctcaaaac
     7321 caaagatctc ctggacttgt atcaacacaa ccctgacctg tgaggtaatg aatggaactg
     7381 accccgaatt aaacctgtat caagatggga aacatctaaa actttctcag agggtcatca
     7441 cacacaagtg gaccaccagc ctgagtgcaa aattcaagtg cacagcaggg aacaaagtca
     7501 gcaaggaatc cagtgtcgag cctgtcagct gtccagagaa aggtctggac atctatctca
     7561 tcattggcat atgtggagga ggcagcctct tgatggtctt tgtggcactg ctcgttttct
     7621 atatcaccaa aaggaaaaaa cagaggagtc ggagaaatga tgaggagctg gagacaagag
     7681 cccacagagt agctactgaa gaaaggggcc ggaagcccca acaaattcca gcttcaaccc
     7741 ctcagaatcc agcaacttcc caacatcctc ctccaccacc tggtcatcgt tcccaggcac
     7801 ctagtcatcg tcccccgcct cctggacacc gtgttcagca ccagcctcag aagaggcctc
     7861 ctgctccgtc gggcacacaa gttcaccagc agaaaggccc gcccctcccc agacctcgag
     7921 ttcagccaaa acctccccat ggggcagcag aaaactcatt gtccccttcc tctaattaaa
     7981 aaagatagaa actgtctttt tcaataaaaa gcactgtgga tttctgcctc tagtcgacga
     8041 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     8101 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga
     8161 gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca
     8221 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg
     8281 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     8341 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc
     8401 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     8461 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     8521 tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa
     8581 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     8641 gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg
     8701 ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc
     8761 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     8821 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     8881 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     8941 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     9001 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     9061 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     9121 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     9181 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     9241 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     9301 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     9361 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     9421 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     9481 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     9541 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     9601 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     9661 tagatccttt tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg
     9721 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     9781 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     9841 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     9901 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     9961 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
    10021 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
    10081 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
    10141 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
    10201 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
    10261 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
    10321 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
    10381 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    10441 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    10501 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    10561 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
    10621 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    10681 gggttccgcg cacatttc
//
