LOCUS       sequence-147472-       14195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NLS-dCas9-GFP11x7-NLS in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14195)
  AUTHORS   Kamiyama D, Sekine S, Barsi-Rhyne B, Hu J, Chen B, Gilbert LA, 
            Ishikawa H, Leonetti MD, Marshall WF, Weissman JS, Huang B
  TITLE     Versatile protein tagging in cells with split fluorescent protein.
  JOURNAL   Nat Commun. 2016 Mar 18;7:11046. doi: 10.1038/ncomms11046.
  PUBMED    26988139
REFERENCE   2  (bases 1 to 14195)
  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: "2016-03-18"; volume: "7:11046. doi"; pages: " 
            10.1038/ncomms11046"
COMMENT     Sequence Label: pHRm-NLS-dCas9-GFP11x7-NLS
FEATURES             Location/Qualifiers
     source          1..14195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..23
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(61..79)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        147..251
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(252..320,321..1112)
                     /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(470..489)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1283..1871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1772..1791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2025..2042
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2112..2132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2117..2446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2297..2432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          3580..3645
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3775..3795
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4220..4354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4257..4276)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4311..4330
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4522..5155
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5202..5327
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5824..6057
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6242..6286
                     /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    6571..6688
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6751..6961
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6820..6955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6977..6997
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      7046..7055
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7052..7078,7079..7081,7082..9568,9569..9571,
                     9572..11155)
                     /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             7052..11155
                     /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             11159..11185
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11165..11191
                     /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             11204..11224
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11231..11251
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11294..11719
                     /codon_start=1
                     /product="seven tandem copies of the 11th beta-strand of 
                     superfolder GFP"
                     /label=GFP11x7
                     /note="each GFP11 associates with GFP(1-10), consisting of 
                     the first 10 beta-strands, to reconstitute fluorescent GFP 
                     (Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSGGRDH
                     MVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSG
                     GRDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGIT"
     CDS             11742..11762
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11772..11828
                     /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             11841..12539
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             12549..12569
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    12591..13179
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12644..12664)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13062..13073)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             13278..13511
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(13536..13554)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(13537..13554)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(13844..13863)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(14077..14096)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
       61 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
      121 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      181 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      241 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      301 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      361 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      421 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      481 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      541 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      601 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      661 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      721 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      781 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      841 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      901 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      961 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1021 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1081 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1141 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1201 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1261 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1321 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1381 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1441 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1501 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1561 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1621 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1681 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1741 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1801 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1861 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1921 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1981 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2041 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2101 aatgcagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2161 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2221 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2281 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2341 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2401 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttggacacaa
     2461 gacaggcttg cgagatatgt ttgagaatac cactttatcc cgcgtcaggg agaggcagtg
     2521 cgtaaaaaga cgcggactca tgtgaaatac tggtttttag tgcgccagat ctctataatc
     2581 tcgcgcaacc tattttcccc tcgaacactt tttaagccgt agataaacag gctgggacac
     2641 ttcacatgag cgaaaaatac atcgtcacct gggacatgtt gcagatccat gcacgtaaac
     2701 tcgcaagccg actgatgcct tctgaacaat ggaaaggcat tattgccgta agccgtggcg
     2761 gtctgtaccg ggtgcgttac tggcgcgtga actgggtatt cgtcatgtcg ataccgtttg
     2821 tatttccagc tacgatcacg acaaccagcg cgagcttaaa gtgctgaaac gcgcagaagg
     2881 cgatggcgaa ggcttcatcg ttattgatga cctggtggat accggtggta ctgcggttgc
     2941 gattcgtgaa atgtatccaa aagcgcactt tgtcaccatc ttcgcaaaac cggctggtcg
     3001 tccgctggtt gatgactatg ttgttgatat cccgcaagat acctggattg aacagccgtg
     3061 ggatatgggc gtcgtattcg tcccgccaat ctccggtcgc taatcttttc aacgcctggc
     3121 actgccgggc gttgttcttt ttaacttcag gcgggttaca atagtttcca gtaagtattc
     3181 tggaggctgc atccatgaca caggcaaacc tgagcgaaac cctgttcaaa ccccgcttta
     3241 aacatcctga aacctcgacg ctagtccgcc gctttaatca cggcgcacaa ccgcctgtgc
     3301 agtcggccct tgatggtaaa accatccctc actggtatcg catgattaac cgtctgatgt
     3361 ggatctggcg cggcattgac ccacgcgaaa tcctcgacgt ccaggcacgt attgtgatga
     3421 gcgatgccga acgtaccgac gatgatttat acgatacggt gattggctac cgtggcggca
     3481 actggattta tgagtgggcc ccggatcttt gtgaaggaac cttacttctg tggtgtgaca
     3541 taattggaca aactacctac agagatttaa agctctaagg taaatataaa atttttaagt
     3601 gtataatgtg ttaaactact gattctaatt gtttgtgtat tttagattcc aacctatgga
     3661 actgatgaat gggagcagtg gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa
     3721 gaaatgccat ctagtgatga tgaggctact gctgactctc aacattctac tcctccaaaa
     3781 aagaagagaa aggtagaaga ccccaaggac tttccttcag aattgctaag ttttttgagt
     3841 catgctgtgt ttagtaatag aactcttgct tgctttgcta tttacaccac aaaggaaaaa
     3901 gctgcactgc tatacaagaa aattatggaa aaatattctg taacctttat aagtaggcat
     3961 aacagttata atcataacat actgtttttt cttactccac acaggcatag agtgtctgct
     4021 attaataact atgctcaaaa attgtgtacc tttagctttt taatttgtaa aggggttaat
     4081 aaggaatatt tgatgtatag tgccttgact agagatcata atcagccata ccacatttgt
     4141 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     4201 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4261 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4321 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa
     4381 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt
     4441 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat
     4501 atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc
     4561 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag
     4621 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     4681 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc
     4741 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag
     4801 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag
     4861 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg
     4921 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct
     4981 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5041 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5101 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     5161 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     5221 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     5281 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     5341 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     5401 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     5461 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     5521 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     5581 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     5641 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     5701 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     5761 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     5821 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     5881 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     5941 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     6001 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     6061 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     6121 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     6181 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     6241 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     6301 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     6361 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     6421 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     6481 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg
     6541 gtatcgccaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     6601 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     6661 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     6721 ttggatcgat aagcttgata tcgaattcag gcaggcagaa gtatgcaaag catgcatctc
     6781 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6841 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6901 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6961 tacgcgcgcc accatgccca agaagaagcg caaggtggga cgcgtctgca ggatatcaag
     7021 cttgcggtac cgcgggcccg ggatcgccac catggacaag aagtacagca tcggcctggc
     7081 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     7141 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggcgc
     7201 cctgctgttc gacagcggag aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     7261 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     7321 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     7381 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     7441 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     7501 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     7561 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     7621 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     7681 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     7741 cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc
     7801 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     7861 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     7921 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     7981 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     8041 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     8101 tttcttcgac cagagcaaga acggctacgc cggctacatc gatggcggag ccagccagga
     8161 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     8221 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     8281 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     8341 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     8401 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     8461 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgcca gcgcccagag
     8521 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     8581 gcacagcctg ctgtacgagt acttcaccgt gtacaacgag ctgaccaaag tgaaatacgt
     8641 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaaag ccatcgtgga
     8701 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     8761 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     8821 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     8881 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     8941 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     9001 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     9061 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     9121 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     9181 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     9241 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     9301 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     9361 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     9421 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     9481 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     9541 caaccggctg tccgactacg atgtggacgc tatcgtgcct cagagctttc tgaaggacga
     9601 ctccatcgat aacaaagtgc tgactcggag cgacaagaac cggggcaaga gcgacaacgt
     9661 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cgccagctgc tgaatgccaa
     9721 gctgattacc cagaggaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     9781 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     9841 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaacg acaaactgat
     9901 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     9961 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
    10021 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
    10081 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
    10141 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
    10201 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaacagg
    10261 cgagatcgtg tgggataagg gccgggactt tgccaccgtg cggaaagtgc tgtctatgcc
    10321 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
    10381 cctgcccaag aggaacagcg acaagctgat cgccagaaag aaggactggg accctaagaa
    10441 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
    10501 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
    10561 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
    10621 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
    10681 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
    10741 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
    10801 ggataatgag cagaaacagc tgtttgtgga acagcacaaa cactacctgg acgagatcat
    10861 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaggt
    10921 gctgagcgcc tacaacaagc acagagacaa gcctatcaga gagcaggccg agaatatcat
    10981 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
    11041 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
    11101 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacgccta
    11161 tccctatgac gtgcccgatt atgccagcct gggcagcggc tcccccaaga aaaaacgcaa
    11221 ggtggaagat cctaagaaaa agcggaaagt ggacggcatt ggtagtggga gcaacggcag
    11281 cagcggatcc atgcgtgacc acatggtcct tcatgagtat gtaaatgctg ctgggattac
    11341 aggtggctct ggaggtagag atcatatggt tctccacgaa tacgttaacg ccgcaggcat
    11401 cactggcggt agtggaggac gcgaccatat ggtactacat gaatatgtca atgcagccgg
    11461 aataaccgga gggtccggag gccgggatca catggtgctg catgagtatg tgaacgcggc
    11521 gggtataact ggtgggtcgg gcggacgaga ccatatggtg cttcacgaat acgtaaacgc
    11581 agctggcatt actggcggat caggtggcag ggatcacatg gtactccatg agtacgtgaa
    11641 cgctgctgga atcacaggcg gtagcggcgg tcgggaccat atggtcctgc acgaatatgt
    11701 caatgctgcc ggtatcaccg cggccgcaag gtggaggtgg acccaagaag aagcgcaagg
    11761 tgggaagcgg agctactaac ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc
    11821 ctggacctgc accaggaagt atgagcgagc tgattaagga gaacatgcac atgaagctgt
    11881 acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc
    11941 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
    12001 ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc
    12061 agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca
    12121 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
    12181 gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc
    12241 agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc
    12301 tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca
    12361 tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact
    12421 atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc
    12481 acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatg
    12541 gtggaggtcc aaaaaagaaa aggaaggtct aacctgcagg gcttatcgat aatcaacctc
    12601 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
    12661 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
    12721 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
    12781 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
    12841 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
    12901 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
    12961 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
    13021 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
    13081 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
    13141 ctcagacgag tcggatctcc ctttgggccg cctccccgca tggtacctgc agggatcgat
    13201 cactgaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
    13261 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
    13321 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
    13381 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
    13441 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
    13501 aatctctagc agcatctaga attaattccg tgtattctat agtgtcacct aaatcgtatg
    13561 tgtatgatac ataaggttat gtattaattg tagccgcgtt ctaacgacaa tatgtacaag
    13621 cctaattgtg tagcatctgg cttactgaag cagaccctat catctctctc gtaaactgcc
    13681 gtcagagtcg gtttggttgg acgaaccttc tgagtttctg gtaacgccgt cccgcacccg
    13741 gaaatggtca gcgaaccaat cagcagggtc atcgctagcc agatcctcta cgccggacgc
    13801 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
    13861 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
    13921 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
    13981 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
    14041 tcgcataagg gagagcgtcg aatggtgcac tctcagtaca atctgctctg atgccgcata
    14101 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
    14161 cccggcatcc gcttacagac aagctgtgac cgtct
//
