LOCUS       sequence-323198-       14638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of dCas9-14xGFP11 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14638)
  AUTHORS   Chen B, Zou W, Xu H, Liang Y, Huang B
  TITLE     Efficient labeling and imaging of protein-coding genes in living 
            cells using CRISPR-Tag.
  JOURNAL   Nat Commun. 2018 Nov 29;9(1):5065. doi: 10.1038/s41467-018-07498-y.
  PUBMED    30498221
REFERENCE   2  (bases 1 to 14638)
  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: "Nat 
            Commun."; date: "2018-11-29"; volume: "9(1):5065. doi"; pages: " 
            10.1038/s41467-018-07498-y"
COMMENT     Sequence Label: BC1209-pHR-dSV40-dCas9-14xGFP11-P2A-BFP
FEATURES             Location/Qualifiers
     source          1..14638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(102..120)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(103..120)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(410..429)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(643..662)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     762..784
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(822..840)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        908..1012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1013..1081,1082..1873)
                     /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(1231..1250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2044..2632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2533..2552
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2786..2803
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2873..2893)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2878..3207
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3058..3193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4341..4406
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4536..4556
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4981..5115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5018..5037)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5072..5091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5283..5916
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5963..6088
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6585..6818
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7003..7047
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7332..7449
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7512..7722
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7581..7716
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7738..7758
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      7807..7816
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7813..7839,7840..7842,7843..10329,10330..10332,
                     10333..11916)
                     /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             7813..11916
                     /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             11920..11946
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11926..11952
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             11965..11985
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11992..12012
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12055..12102
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12118..12165
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12181..12228
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12244..12291
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12307..12354
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12370..12417
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12433..12480
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12496..12543
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12559..12606
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12622..12669
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12685..12732
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12748..12795
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12811..12858
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12874..12921
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             12946..12966
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12976..13032
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             13045..13743
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             13753..13773
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    13795..14383
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13848..13868)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14266..14277)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             join(14482..14638,1..77)
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
       61 gtggaaaatc tctagcagca tctagaatta attccgtgta ttctatagtg tcacctaaat
      121 cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg
      181 tacaagccta attgtgtagc atctggctta ctgaagcaga ccctatcatc tctctcgtaa
      241 actgccgtca gagtcggttt ggttggacga accttctgag tttctggtaa cgccgtcccg
      301 cacccggaaa tggtcagcga accaatcagc agggtcatcg ctagccagat cctctacgcc
      361 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
      421 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
      481 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
      541 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
      601 caggagtcgc ataagggaga gcgtcgaatg gtgcactctc agtacaatct gctctgatgc
      661 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      721 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
      781 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
      841 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
      901 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      961 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1021 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1081 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1141 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1201 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1261 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1321 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1381 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1441 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1501 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1561 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1621 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1681 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1741 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1801 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1861 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1921 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1981 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2041 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2101 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2161 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2221 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2281 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2341 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2401 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2461 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2521 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2581 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2641 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2701 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2761 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2821 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct gtggaatgtg
     2881 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2941 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3001 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3061 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3121 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3181 ttttttggag gcctaggctt ttgcaaaaag cttggacaca agacaggctt gcgagatatg
     3241 tttgagaata ccactttatc ccgcgtcagg gagaggcagt gcgtaaaaag acgcggactc
     3301 atgtgaaata ctggttttta gtgcgccaga tctctataat ctcgcgcaac ctattttccc
     3361 ctcgaacact ttttaagccg tagataaaca ggctgggaca cttcacatga gcgaaaaata
     3421 catcgtcacc tgggacatgt tgcagatcca tgcacgtaaa ctcgcaagcc gactgatgcc
     3481 ttctgaacaa tggaaaggca ttattgccgt aagccgtggc ggtctgtacc gggtgcgtta
     3541 ctggcgcgtg aactgggtat tcgtcatgtc gataccgttt gtatttccag ctacgatcac
     3601 gacaaccagc gcgagcttaa agtgctgaaa cgcgcagaag gcgatggcga aggcttcatc
     3661 gttattgatg acctggtgga taccggtggt actgcggttg cgattcgtga aatgtatcca
     3721 aaagcgcact ttgtcaccat cttcgcaaaa ccggctggtc gtccgctggt tgatgactat
     3781 gttgttgata tcccgcaaga tacctggatt gaacagccgt gggatatggg cgtcgtattc
     3841 gtcccgccaa tctccggtcg ctaatctttt caacgcctgg cactgccggg cgttgttctt
     3901 tttaacttca ggcgggttac aatagtttcc agtaagtatt ctggaggctg catccatgac
     3961 acaggcaaac ctgagcgaaa ccctgttcaa accccgcttt aaacatcctg aaacctcgac
     4021 gctagtccgc cgctttaatc acggcgcaca accgcctgtg cagtcggccc ttgatggtaa
     4081 aaccatccct cactggtatc gcatgattaa ccgtctgatg tggatctggc gcggcattga
     4141 cccacgcgaa atcctcgacg tccaggcacg tattgtgatg agcgatgccg aacgtaccga
     4201 cgatgattta tacgatacgg tgattggcta ccgtggcggc aactggattt atgagtgggc
     4261 cccggatctt tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta
     4321 cagagattta aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac
     4381 tgattctaat tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt
     4441 ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga agaaatgcca tctagtgatg
     4501 atgaggctac tgctgactct caacattcta ctcctccaaa aaagaagaga aaggtagaag
     4561 accccaagga ctttccttca gaattgctaa gttttttgag tcatgctgtg tttagtaata
     4621 gaactcttgc ttgctttgct atttacacca caaaggaaaa agctgcactg ctatacaaga
     4681 aaattatgga aaaatattct gtaaccttta taagtaggca taacagttat aatcataaca
     4741 tactgttttt tcttactcca cacaggcata gagtgtctgc tattaataac tatgctcaaa
     4801 aattgtgtac ctttagcttt ttaatttgta aaggggttaa taaggaatat ttgatgtata
     4861 gtgccttgac tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     4921 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     4981 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5041 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5101 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc
     5161 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
     5221 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
     5281 gttggaaggg ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca
     5341 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     5401 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     5461 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     5521 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     5581 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     5641 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     5701 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     5761 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5821 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5881 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     5941 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     6001 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     6061 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6121 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     6181 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     6241 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     6301 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     6361 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     6421 gcaagcggcc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     6481 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     6541 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     6601 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     6661 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     6721 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     6781 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     6841 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     6901 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     6961 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     7021 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     7081 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     7141 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     7201 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     7261 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcca aatggcagta
     7321 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     7381 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     7441 caaaattttc gggtttatta cagggacagc agagatccag tttggatcga taagcttgat
     7501 atcgaattca ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     7561 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     7621 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     7681 tattccagaa gtagtgagga ggcttttttg gaggcctagg ctacgcgcgc caccatgccc
     7741 aagaagaagc gcaaggtggg acgcgtctgc aggatatcaa gcttgcggta ccgcgggccc
     7801 gggatcgcca ccatggacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg
     7861 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     7921 aacaccgacc ggcacagcat caagaagaac ctgatcggcg ccctgctgtt cgacagcgga
     7981 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     8041 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     8101 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     8161 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     8221 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     8281 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     8341 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     8401 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     8461 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     8521 ctgttcggca acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     8581 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     8641 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     8701 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     8761 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     8821 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     8881 aacggctacg ccggctacat cgatggcgga gccagccagg aagagttcta caagttcatc
     8941 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     9001 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     9061 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     9121 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     9181 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     9241 aacttcgagg aagtggtgga caagggcgcc agcgcccaga gcttcatcga gcggatgacc
     9301 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     9361 tacttcaccg tgtacaacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     9421 cccgccttcc tgagcggcga gcagaaaaaa gccatcgtgg acctgctgtt caagaccaac
     9481 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     9541 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     9601 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     9661 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     9721 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     9781 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     9841 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     9901 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     9961 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
    10021 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
    10081 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
    10141 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
    10201 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
    10261 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
    10321 gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg actccatcga taacaaagtg
    10381 ctgactcgga gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
    10441 aagaagatga agaactactg gcgccagctg ctgaatgcca agctgattac ccagaggaag
    10501 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
    10561 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
    10621 tcccggatga acactaagta cgacgagaac gacaaactga tccgggaagt gaaagtgatc
    10681 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
    10741 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
    10801 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
    10861 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
    10921 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
    10981 atccggaagc ggcctctgat cgagacaaac ggcgaaacag gcgagatcgt gtgggataag
    11041 ggccgggact ttgccaccgt gcggaaagtg ctgtctatgc cccaagtgaa tatcgtgaaa
    11101 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
    11161 gacaagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
    11221 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
    11281 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
    11341 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
    11401 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
    11461 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
    11521 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
    11581 ctgtttgtgg aacagcacaa acactacctg gacgagatca tcgagcagat cagcgagttc
    11641 tccaagagag tgatcctggc cgacgctaat ctggacaagg tgctgagcgc ctacaacaag
    11701 cacagagaca agcctatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
    11761 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
    11821 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
    11881 gagacacgga tcgacctgtc tcagctggga ggcgacgcct atccctatga cgtgcccgat
    11941 tatgccagcc tgggcagcgg ctcccccaag aaaaaacgca aggtggaaga tcctaagaaa
    12001 aagcggaaag tggacggcat tggtagtggg agcaacggca gcagcggatc catgcgggac
    12061 cacatggtgc tgcatgaata cgtgaacgcg gcggggataa cgggtgggtc cggaggtaga
    12121 gaccatatgg ttctccacga gtacgtcaac gcagcgggca ttacaggcgg ttcaggtggc
    12181 agggatcaca tggtgttaca tgaatatgta aatgcggctg gaatcaccgg aggatctgga
    12241 ggccgggacc atatggtctt gcacgagtac gttaacgctg ctggtataac cgggggctcg
    12301 ggcggtagag atcacatggt actccatgag tatgtcaatg ccgcgggtat aactggtggg
    12361 tcaggtggac gcgaccatat ggttttacat gaatatgtga acgcagcagg aattactggc
    12421 ggttctgggg gtagggatca tatggtacta cacgagtacg tgaacgccgc tggcatcacc
    12481 ggcggatcag ggggccgcga ccatatggtt ctccacgaat acgtgaatgc cgccggaatt
    12541 acaggcggat cgggaggtcg tgatcacatg gtactacatg agtacgtaaa cgcggcgggg
    12601 atcacgggag gatctggtgg tcgagaccat atggtgcttc acgagtatgt taacgcagca
    12661 ggtattaccg gcggttctgg tggcagagac catatggtcc tgcatgagta tgtcaatgcc
    12721 gctggcatca ctggaggctc aggtggaaga gatcacatgg ttctacacga atatgtgaat
    12781 gcagccggta tcactggtgg gtcaggaggg cgcgaccaca tggtactcca tgaatacgtt
    12841 aatgcagctg ggattacagg gggttccggc ggtcgggatc atatggtttt gcatgagtac
    12901 gtcaacgctg ccggaatcac acagcggccg caaggtggag gtggacccaa gaagaagcgc
    12961 aaggtgggaa gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag
    13021 aaccctggac ctgcaccagg aagtatgagc gagctgatta aggagaacat gcacatgaag
    13081 ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc
    13141 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
    13201 ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac
    13261 acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga
    13321 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
    13381 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg
    13441 atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc
    13501 ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca
    13561 aacatcaaga ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc
    13621 tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag
    13681 cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt
    13741 aatggtggag gtccaaaaaa gaaaaggaag gtctaacctg cagggcttat cgataatcaa
    13801 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
    13861 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
    13921 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
    13981 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
    14041 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
    14101 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
    14161 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
    14221 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
    14281 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
    14341 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatggtac ctgcagggat
    14401 cgatcactga ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
    14461 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
    14521 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
    14581 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagt
//
