LOCUS       sequence-354783-       15344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes S. pyogenes dCas9 with n-terminal fusion of human GCN5 core 
            (aa 473-676) and c-terminal fusion of human p300 core (aa 1048-1664)
            driven by EF-1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15344)
  AUTHORS   Wang K, Escobar M, Li J, Mahata B, Goell J, Shah S, Cluck M, Hilton 
            IB
  TITLE     Systematic comparison of CRISPR-based transcriptional activators 
            uncovers gene-regulatory features of enhancer-promoter interactions.
  JOURNAL   Nucleic Acids Res. 2022 Aug 12;50(14):7842-7855. doi: 
            10.1093/nar/gkac582.
  PUBMED    35849129
REFERENCE   2  (bases 1 to 15344)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2022-08-12- 12"; volume: "50"; issue: 
            "14"; pages: "7842-7855"
COMMENT     Sequence Label: GCN5 core-dCas9-p300 core
FEATURES             Location/Qualifiers
     source          1..15344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(344..355)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             533..766
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(792..809)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    798..1022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1068..1496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1155..1174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1365..1386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1505..1525)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1510..1839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1690..1825
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1752..1771
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1887..1934
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1953..2327
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2457..2578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2494..2513)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2548..2567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2627..2643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2627..2643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2640..2662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2651..2667
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2675..2681,2682..2699,2700..2705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2720..2741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2858..2875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3029..3617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3109..3128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3788..4579,4580..4648))
                     /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     4411..4430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4649..4753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4828..4847)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5019..5398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5400..5598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5553..5573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5616..5796
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5843..5968
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6461..6694
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6879..6923
                     /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    7221..7338
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        7470..7545
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7607..7818
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             8461..8481
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      8494..8503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(8500..8526,8527..8529,8530..11016,11017..11019,
                     11020..12603)
                     /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             8500..12603
                     /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             12604..12651
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12658..14508
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      14272..14281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             14515..14538
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             14548..14604
                     /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             14605..15201
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     15098..15118
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    join(15217..15344,1..461)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(15270..15290)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
       61 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
      121 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
      181 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
      241 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
      301 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
      361 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
      421 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg
      481 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
      541 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
      601 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
      661 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
      721 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac
      781 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
      841 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
      901 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
      961 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1021 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1081 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1141 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1201 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1261 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1321 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1381 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1441 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1501 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1561 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1621 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     1681 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     1741 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     1801 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     1861 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     1921 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     1981 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     2041 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     2101 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     2161 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     2221 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     2281 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     2341 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2401 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     2461 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2521 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2581 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     2641 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2701 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2761 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2821 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2881 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2941 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3001 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3061 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3781 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3841 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3901 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3961 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4021 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4081 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4141 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4201 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4261 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4321 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4381 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4441 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4501 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4561 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4621 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4681 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4741 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     4801 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4861 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     4921 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     4981 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     5041 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     5101 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     5161 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     5221 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     5281 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     5341 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     5401 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     5461 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     5521 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     5581 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     5641 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5701 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5761 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     5821 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     5881 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     5941 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6001 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     6061 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     6121 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     6181 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     6241 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     6301 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     6361 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     6421 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     6481 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     6541 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     6601 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     6661 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     6721 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     6781 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     6841 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     6901 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     6961 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     7021 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     7081 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     7141 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
     7201 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     7261 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     7321 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt
     7381 aaggtaccga tatcactagt gttccattaa ctgcgctaaa aataattttt aaatcttttt
     7441 tagcttcttg ctcttttttg tacgtctctg ttttagagct agaaatagca agttaaaata
     7501 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgaattcgct
     7561 agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca
     7621 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg tgcctagaga
     7681 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     7741 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     7801 tttgccgcca gaacacagga ccggttctag agcgccacca tgctggggcc tgagacgagc
     7861 ctgctttcgg ccaatgcggc ccgggatgag acagcccgcc tggaggagcg ccgcggcatc
     7921 atcgagttcc atgtcatcgg caactcactg acgcccaagg ccaaccggcg ggtgttgctg
     7981 tggctcgtgg ggctgcagaa tgtcttttcc caccagctgc cgcgcatgcc taaggagtat
     8041 atcgcccgcc tcgtctttga cccgaagcac aagactctgg ccttgatcaa ggatgggcgg
     8101 gtcatcggtg gcatctgctt ccgcatgttt cccacccagg gcttcacgga gattgtcttc
     8161 tgtgctgtca cctcgaatga gcaggtcaag ggttatggga cccacctgat gaaccacctg
     8221 aaggagtatc acatcaagca caacattctc tacttcctca cctacgccga cgagtacgcc
     8281 atcggctact tcaaaaagca gggtttctcc aaggacatca aggtgcccaa gagccgctac
     8341 ctgggctaca tcaaggacta cgagggagcg acgctgatgg agtgtgagct gaatccccgc
     8401 atcccctaca cggagctgtc ccacatcatc aagaagcaga aagagatcat caagggatcc
     8461 cccaagaaga agaggaaggt gggatccagc gctgccacca tggacaagaa gtacagcatc
     8521 ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     8581 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     8641 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
     8701 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     8761 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     8821 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     8881 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     8941 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     9001 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     9061 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     9121 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     9181 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     9241 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     9301 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     9361 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     9421 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     9481 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     9541 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     9601 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     9661 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     9721 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     9781 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     9841 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     9901 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     9961 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
    10021 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
    10081 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
    10141 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
    10201 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
    10261 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
    10321 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
    10381 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
    10441 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
    10501 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
    10561 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
    10621 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
    10681 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
    10741 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
    10801 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
    10861 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
    10921 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
    10981 ctggacatca accggctgtc cgactacgat gtggacgcca tcgtgcctca gagctttctg
    11041 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
    11101 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
    11161 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
    11221 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
    11281 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
    11341 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
    11401 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
    11461 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
    11521 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
    11581 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
    11641 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
    11701 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
    11761 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
    11821 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
    11881 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
    11941 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
    12001 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
    12061 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
    12121 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
    12181 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
    12241 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
    12301 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
    12361 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
    12421 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
    12481 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
    12541 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
    12601 gacaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa aggatccatt
    12661 ttcaaaccag aagaactacg acaggcactg atgccaactt tggaggcact ttaccgtcag
    12721 gatccagaat cccttccctt tcgtcaacct gtggaccctc agcttttagg aatccctgat
    12781 tactttgata ttgtgaagag ccccatggat ctttctacca ttaagaggaa gttagacact
    12841 ggacagtatc aggagccctg gcagtatgtc gatgatattt ggcttatgtt caataatgcc
    12901 tggttatata accggaaaac atcacgggta tacaaatact gctccaagct ctctgaggtc
    12961 tttgaacaag aaattgaccc agtgatgcaa agccttggat actgttgtgg cagaaagttg
    13021 gagttctctc cacagacact gtgttgctac ggcaaacagt tgtgcacaat acctcgtgat
    13081 gccacttatt acagttacca gaacaggtat catttctgtg agaagtgttt caatgagatc
    13141 caaggggaga gcgtttcttt gggggatgac ccttcccagc ctcaaactac aataaataaa
    13201 gaacaatttt ccaagagaaa aaatgacaca ctggatcctg aactgtttgt tgaatgtaca
    13261 gagtgcggaa gaaagatgca tcagatctgt gtccttcacc atgagatcat ctggcctgct
    13321 ggattcgtct gtgatggctg tttaaagaaa agtgcacgaa ctaggaaaga aaataagttt
    13381 tctgctaaaa ggttgccatc taccagactt ggcacctttc tagagaatcg tgtgaatgac
    13441 tttctgaggc gacagaatca ccctgagtca ggagaggtca ctgttagagt agttcatgct
    13501 tctgacaaaa ccgtggaagt aaaaccaggc atgaaagcaa ggtttgtgga cagtggagag
    13561 atggcagaat cctttccata ccgaaccaaa gccctctttg cctttgaaga aattgatggt
    13621 gttgacctgt gcttctttgg catgcatgtt caagagtatg gctctgactg ccctccaccc
    13681 aaccagagga gagtatacat atcttacctc gatagtgttc atttcttccg tcctaaatgc
    13741 ttgaggactg cagtctatca tgaaatccta attggatatt tagaatatgt caagaaatta
    13801 ggttacacaa cagggcatat ttgggcatgt ccaccaagtg agggagatga ttatatcttc
    13861 cattgccatc ctcctgacca gaagataccc aagcccaagc gactgcagga atggtacaaa
    13921 aaaatgcttg acaaggctgt atcagagcgt attgtccatg actacaagga tatttttaaa
    13981 caagctactg aagatagatt aacaagtgca aaggaattgc cttatttcga gggtgatttc
    14041 tggcccaatg ttctggaaga aagcattaag gaactggaac aggaggaaga agagagaaaa
    14101 cgagaggaaa acaccagcaa tgaaagcaca gatgtgacca agggagacag caaaaatgct
    14161 aaaaagaaga ataataagaa aaccagcaaa aataagagca gcctgagtag gggcaacaag
    14221 aagaaacccg ggatgcccaa tgtatctaac gacctctcac agaaactata tgccaccatg
    14281 gagaagcata aagaggtctt ctttgtgatc cgcctcattg ctggccctgc tgccaactcc
    14341 ctgcctccca ttgttgatcc tgatcctctc atcccctgcg atctgatgga tggtcgggat
    14401 gcgtttctca cgctggcaag ggacaagcac ctggagttct cttcactccg aagagcccag
    14461 tggtccacca tgtgcatgct ggtggagctg cacacgcaga gccaggacgg atccgattac
    14521 aaagacgatg acgataaggg atccggcgca acaaacttct ctctgctgaa acaagccgga
    14581 gatgtcgaag agaatcctgg accgaccgag tacaagccca cggtgcgcct cgccacccgc
    14641 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
    14701 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
    14761 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
    14821 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
    14881 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
    14941 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac
    15001 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
    15061 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
    15121 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
    15181 acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt
    15241 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
    15301 gctttaatgc ctttgtatca tgctattgct tcccgtatgg cttt
//
