LOCUS       sequence-147542-       11395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-GCN4 under actin promoter for SunTag CRISPRa in 
            Drosophila cell culture.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11395)
  AUTHORS   Chavez A, Tuttle M, Pruitt BW, Ewen-Campen B, Chari R, Ter-Ovanesyan
            D, Haque SJ, Cecchi RJ, Kowal EJ, Buchthal J, Housden BE, Perrimon 
            N, Collins JJ, Church G
  TITLE     Comparison of Cas9 activators in multiple species.
  JOURNAL   Nat Methods. 2016 May 23. doi: 10.1038/nmeth.3871.
  PUBMED    27214048
REFERENCE   2  (bases 1 to 11395)
  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 
            Methods. 2016 May 23. doi: 10.1038/nmeth.3871."
COMMENT     Sequence Label: pAct:dCas9-GCN4
FEATURES             Location/Qualifiers
     source          1..11395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        140..2649
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     protein_bind    2667..2691
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2706..2726
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      2775..2784
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2781..2807,2808..2810,2811..5297,5298..5300,
                     5301..6884)
                     /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             2781..6884
                     /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             6888..6914
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6894..6920
                     /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             6933..6953
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6960..6980
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7044..7100
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7116..7172
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7188..7244
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7260..7316
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7332..7388
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7404..7460
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7476..7532
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7548..7604
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7620..7676
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7692..7748
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             7830..7850
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(7864..7888)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      7896..7905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7902..7904,7905..7907,7908..8618)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7902..7904,7905..7907,7908..8618)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8208..8227)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8555..8576
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          8723..8788
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8918..8938
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    9363..9497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9400..9419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9454..9473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(9594..9662,9663..10454)
                     /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(9812..9831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10625..11213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11114..11133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11367..11384
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggcaact
       61 cgtgaaaggt aggcggatca gcggccgcag catgcaattc tatattctaa aaacacaaat
      121 gatacttcta aaaaaaaatc atgaatggca tcaactctga atcaaatctt tgcagatgca
      181 cctacttctc atttccactg tcacatcatt tttccagatc tcgctgcctg ttatgtggcc
      241 cacaaaccaa gacacgtttt atggccatta aagctggctg atcgtcgcca aacaccaaat
      301 acataatgaa tatgtacaca ttcgagaaag aagcgatcaa agaagcgtct tcgggcggag
      361 taggagaatg cggaggagaa ggagaacgag ctgatctagt atctctccac aatccaatgc
      421 caactgacca actggccata ttcggagcaa tttgaagcca atttccatcg cctggcgatc
      481 gctccattct tggctatatg tttttcaccg ttacccgggg ccattttcaa agactcgtcg
      541 gcaagataag attgtgtcac tcgctgtctc tcttcatttg tcgaagaatg ctgaggaatt
      601 tcgcgatgac gtcggcgagt attttgaaga atgagaataa tttgtattta tacgaaaatc
      661 agttagtgga attttctaca aaaacatgtt atctatagat aattttgttg caaaatatgt
      721 tgactatgac aaagattgta tgtatatacc tttaatgtat tctcattttc ttatgtattt
      781 ataatggcaa tgatgatact gatgatattt taagatgatg ccagaccaaa aggcttgaat
      841 ttctgcgtct tttgccgaac gcagtgcatg tgcaattgtt gttttttgga atattcaatt
      901 ttcggactgt ccgctttgat ttcagtttct tggcttattc aaaaagcaaa gtaaagccaa
      961 aaaagcgaga tggcaatacc aaatgcggca aaacggtagt ggaaggaaag gggtgcgggg
     1021 cagcggaagg aagggtgggg cggggcgtgg cggggtctgt ggctgggcgc gacgtcaccg
     1081 acgttggagc cactcctttg accatgtgtg cgtgtgtgta ttattcgtgt ctcgccactc
     1141 gccggttgtt tttttctttt tatgctgcgc tctctctagc gccatctcgc ttacgcatgc
     1201 tcaacgcacc gcatgttgcc gtttcctttt atgcgtcatt ttggctcgaa ataggcaatt
     1261 atttaaacaa agattagtca acgaaaacgc taaaataaat aagtctacaa tatggttact
     1321 tattgccatg tgtgtgcagc caacgatagc aacaaaagca acaacacagg tggctttccc
     1381 tctttcactt tttgtttgca agccgcgtgc gagcaagacg gcacgaccgg caaacgcaat
     1441 tacgctgaca aagagcagac gaagttttgg cgaaaaacat caaggcgcct gatacgaatg
     1501 catttgcaat aacaattgcg atatttaata ttgtttatga agctgtttga cttcaaaaca
     1561 cacaaaaaaa aaaataaaac aaattatttg aaagagaatt aggaatcgga cgcttatcgt
     1621 tagggtaaca acaagaaatg cttactgagt cacagcctct ggaaaactgc cgcaagccag
     1681 agagagagag aaaaagaggg agagcagctt agaccgcatg tgcttgtgtg tgaggcgtct
     1741 ctctcttcgt ctctgttgcg caaacgcata gactgcactg agaaaatcga ttacctattt
     1801 tttatgaatg aatatttgca ctattactat tcaaaactat taagatagca atcacattca
     1861 atagccaaat actataccac ctgagcgatg caacgaaatg atcaatttga gcaaaaatgc
     1921 tgcatattta ggacggcatc attatagaaa tgcttcttgc tgtgtacttt tctctcgtct
     1981 ggcagctgtt tcgccgttat tgttaaaacc ggcttaagtt aggtgtgttt tctacgacta
     2041 gtgaatgccc tactagaaga tgtgtgttgc acaaaatgtc cctggaataa ccaatttgaa
     2101 gtgcagatag cagtaaacgt aagctaatat gaatattatt taactgtaat gttttaatat
     2161 cgctggacat tactaataaa cccactataa acacatgtac atatgtatgt tttggcatac
     2221 aatgagtagt tggggaaaaa atgtgtaaaa gcaccgtgac catcacagca taaagataac
     2281 cagctgaagt atcgaatatg agtaaccccc aaattgaatc acatgccgca actgatagga
     2341 cccatggaag tacactcttc atggcgatat acaagacaca cacaagcacg aacacccagt
     2401 tgcggaggaa attctccgta aatgaaaacc caatcggcga acaattcata cccatatatg
     2461 gtaaaagttt tgaacgcgac ttgagagcgg agagcattgc ggctgataag gttttagcgc
     2521 taagcgggct ttataaaacg ggctgcggga ccagttttca tatcactacc gtttgagttc
     2581 ttgtgctgtg tggatactcc tcccgacaca aagccgctcc atcagccagc agtcgtctaa
     2641 tccagagacc ccggatccag atatcgacaa gtttgtacaa aaaagcaggc tacaaagcca
     2701 ccatgcccaa gaagaagcgc aaggtgggac gcgtctgcag gatatcaagc ttgcggtacc
     2761 gcgggcccgg gatcgccacc atggacaaga agtacagcat cggcctggcc atcggcacca
     2821 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
     2881 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggcgcc ctgctgttcg
     2941 acagcggaga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
     3001 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
     3061 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
     3121 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     3181 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
     3241 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
     3301 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     3361 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     3421 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     3481 agaatggcct gttcggcaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     3541 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     3601 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     3661 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     3721 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     3781 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     3841 agagcaagaa cggctacgcc ggctacatcg atggcggagc cagccaggaa gagttctaca
     3901 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     3961 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     4021 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     4081 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     4141 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     4201 ccccctggaa cttcgaggaa gtggtggaca agggcgccag cgcccagagc ttcatcgagc
     4261 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     4321 tgtacgagta cttcaccgtg tacaacgagc tgaccaaagt gaaatacgtg accgagggaa
     4381 tgagaaagcc cgccttcctg agcggcgagc agaaaaaagc catcgtggac ctgctgttca
     4441 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     4501 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     4561 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     4621 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     4681 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     4741 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     4801 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     4861 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     4921 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     4981 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
     5041 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
     5101 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
     5161 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
     5221 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
     5281 ccgactacga tgtggacgct atcgtgcctc agagctttct gaaggacgac tccatcgata
     5341 acaaagtgct gactcggagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
     5401 aggtcgtgaa gaagatgaag aactactggc gccagctgct gaatgccaag ctgattaccc
     5461 agaggaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
     5521 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
     5581 tcctggactc ccggatgaac actaagtacg acgagaacga caaactgatc cgggaagtga
     5641 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
     5701 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
     5761 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
     5821 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
     5881 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
     5941 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaacaggc gagatcgtgt
     6001 gggataaggg ccgggacttt gccaccgtgc ggaaagtgct gtctatgccc caagtgaata
     6061 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
     6121 ggaacagcga caagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
     6181 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
     6241 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
     6301 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
     6361 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
     6421 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
     6481 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
     6541 agaaacagct gtttgtggaa cagcacaaac actacctgga cgagatcatc gagcagatca
     6601 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaggtg ctgagcgcct
     6661 acaacaagca cagagacaag cctatcagag agcaggccga gaatatcatc cacctgttta
     6721 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
     6781 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
     6841 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacgcctat ccctatgacg
     6901 tgcccgatta tgccagcctg ggcagcggct cccccaagaa aaaacgcaag gtggaagatc
     6961 ctaagaaaaa gcggaaagtg gacggcattg gtagtgggag caacggcagc agcggatcca
     7021 acggtccgac tgacgccgcg gaagaagaac ttttgagcaa gaattatcat cttgagaacg
     7081 aagtggctcg tcttaagaaa ggttctggca gtggagaaga actgctttca aagaattacc
     7141 acctggaaaa tgaggtagct agactgaaaa aggggagcgg aagtggggag gagttgctga
     7201 gcaaaaatta tcatttggag aacgaagtag cacgactaaa gaaagggtcc ggatcgggtg
     7261 aggagttact ctcgaaaaat tatcatctcg aaaacgaagt ggctcggcta aaaaagggca
     7321 gtggttctgg agaagagcta ttatctaaaa actaccacct cgaaaatgag gtggcacgct
     7381 taaaaaaggg aagtggcagt ggtgaagagc tactatccaa gaattatcat cttgagaacg
     7441 aggtagcgcg tttgaagaag ggttccggct caggagagga actgctctcg aagaactatc
     7501 atcttgaaaa tgaggtcgct cgattaaaaa agggatcggg cagtggtgag gaactacttt
     7561 caaagaatta ccacctcgaa aacgaagtag ctcgattaaa gaaaggttca gggtcgggtg
     7621 aagaattact gagtaaaaat tatcatctgg aaaatgaggt agcgagacta aaaaagggga
     7681 gtggttctgg cgaggaattg ctatcgaaaa attatcatct tgagaacgaa gttgctaggc
     7741 tcaaaaaggg ctcaggctca ggcaccgcgg taaacatagg tggtggaacc ggtccgatgg
     7801 atctacagcg gccgcaaggt ggaggtggac ccaagaagaa gcgcaaggtg ggaagcggat
     7861 aaaacccagc tttcttgtac aaagtggtga gctccgccac catggtgagc aagggcgagg
     7921 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     7981 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     8041 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     8101 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     8161 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     8221 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     8281 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     8341 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     8401 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     8461 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     8521 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     8581 ccgccgggat cactctcggc atggacgagc tgtacaagca ccggtccacg tgacgtaagc
     8641 tagcaggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg acaaactacc
     8701 tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat gtgttaaact
     8761 actgattcta attgtttgtg tattttagat tccaacctat ggaactgatg aatgggagca
     8821 gtggtggaat gcctttaatg aggaaaacct gttttgctca gaagaaatgc catctagtga
     8881 tgatgaggct actgctgact ctcaacattc tactcctcca aaaaagaaga gaaaggtaga
     8941 agaccccaag gactttcctt cagaattgct aagttttttg agtcatgctg tgtttagtaa
     9001 tagaactctt gcttgctttg ctatttacac cacaaaggaa aaagctgcac tgctatacaa
     9061 gaaaattatg gaaaaatatt ctgtaacctt tataagtagg cataacagtt ataatcataa
     9121 catactgttt tttcttactc cacacaggca tagagtgtct gctattaata actatgctca
     9181 aaaattgtgt acctttagct ttttaatttg taaaggggtt aataaggaat atttgatgta
     9241 tagtgccttg actagagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     9301 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     9361 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     9421 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     9481 cttatcatgt ctggatcccg tttaaactac gcgtaattca aacagggttc tggcgtcgtt
     9541 ctcgtactgt tttccccagg ccagtgcttt agcgttattg aaaaaggaag agtatgagta
     9601 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     9661 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     9721 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     9781 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     9841 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     9901 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     9961 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
    10021 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
    10081 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
    10141 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
    10201 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
    10261 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
    10321 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
    10381 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
    10441 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
    10501 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
    10561 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
    10621 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
    10681 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
    10741 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
    10801 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
    10861 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
    10921 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
    10981 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
    11041 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
    11101 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
    11161 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
    11221 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
    11281 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
    11341 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaaga
//
