LOCUS       sequence-354777-       13448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes S. pyogenes dCas9 with c-terminal fusion of human GCN5 core 
            (aa 473-676) driven by EF-1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13448)
  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 13448)
  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: dCas9-GCN5 core
FEATURES             Location/Qualifiers
     source          1..13448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     51..70
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(130..146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(130..146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(143..165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    154..170
                     /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(178..184,185..202,203..208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    223..244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(361..378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(532..1120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(612..631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1291..2082,2083..2151))
                     /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     1914..1933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2152..2256)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2331..2350)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2522..2901
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2903..3101
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3056..3076
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3119..3299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3346..3471
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3964..4197
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4382..4426
                     /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    4724..4841
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        4973..5048
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5110..5321
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5340..5349
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5346..5372,5373..5375,5376..7862,7863..7865,
                     7866..9449)
                     /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             5346..9449
                     /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             9450..9497
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10122..10145
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10155..10211
                     /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             10212..10808
                     /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     10705..10725
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10824..11412
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10877..10897)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11295..11306)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11484..11717
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11743..11760)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11749..11973
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12019..12447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12106..12125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12316..12337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12456..12476)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12461..12790
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12641..12776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12703..12722
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12838..12885
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12904..13278
                     /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    join(13408..13448,1..81)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(13445..13448,1..16))
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
       61 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
      121 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
      181 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
      241 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
      301 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
      361 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      421 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      481 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      541 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      601 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      661 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      721 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      781 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      841 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      901 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      961 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1021 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2281 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     2341 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     2401 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     2461 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     2521 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     2581 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     2641 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     2701 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     2761 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     2821 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     2881 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     2941 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     3001 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     3061 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
     3121 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     3181 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     3241 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     3301 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     3361 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     3421 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     3481 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     3541 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     3601 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     3661 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     3721 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     3781 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     3841 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     3901 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     3961 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     4021 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     4081 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     4141 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     4201 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     4261 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     4321 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     4381 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     4441 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     4501 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     4561 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     4621 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     4681 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     4741 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     4801 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     4861 gcagagatcc agtttggtta attaaggtac cgatatcact agtgttccat taactgcgct
     4921 aaaaataatt tttaaatctt ttttagcttc ttgctctttt ttgtacgtct ctgttttaga
     4981 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     5041 gtcggtgctt ttttgaattc gctagctagg tcttgaaagg agtgggaatt ggctccggtg
     5101 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     5161 gcaattgatc cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     5221 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     5281 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggaccggttc tagagcgctg
     5341 ccaccatgga caagaagtac agcatcggcc tggccatcgg caccaactct gtgggctggg
     5401 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
     5461 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
     5521 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
     5581 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
     5641 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
     5701 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
     5761 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
     5821 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
     5881 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
     5941 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga
     6001 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg
     6061 gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg
     6121 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc
     6181 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg
     6241 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg
     6301 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg
     6361 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct
     6421 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca
     6481 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     6541 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc
     6601 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     6661 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
     6721 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
     6781 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
     6841 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
     6901 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
     6961 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
     7021 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
     7081 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
     7141 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
     7201 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
     7261 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
     7321 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
     7381 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg
     7441 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
     7501 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     7561 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     7621 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     7681 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     7741 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     7801 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     7861 acgccatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
     7921 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     7981 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     8041 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     8101 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     8161 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     8221 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     8281 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     8341 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     8401 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     8461 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     8521 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     8581 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     8641 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
     8701 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
     8761 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     8821 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
     8881 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
     8941 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
     9001 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
     9061 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
     9121 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
     9181 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
     9241 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
     9301 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
     9361 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
     9421 ggatcgacct gtctcagctg ggaggcgaca agcgacctgc cgccacaaag aaggctggac
     9481 aggctaagaa gaagaaagga tccctggggc ctgagacgag cctgctttcg gccaatgcgg
     9541 cccgggatga gacagcccgc ctggaggagc gccgcggcat catcgagttc catgtcatcg
     9601 gcaactcact gacgcccaag gccaaccggc gggtgttgct gtggctcgtg gggctgcaga
     9661 atgtcttttc ccaccagctg ccgcgcatgc ctaaggagta tatcgcccgc ctcgtctttg
     9721 acccgaagca caagactctg gccttgatca aggatgggcg ggtcatcggt ggcatctgct
     9781 tccgcatgtt tcccacccag ggcttcacgg agattgtctt ctgtgctgtc acctcgaatg
     9841 agcaggtcaa gggttatggg acccacctga tgaaccacct gaaggagtat cacatcaagc
     9901 acaacattct ctacttcctc acctacgccg acgagtacgc catcggctac ttcaaaaagc
     9961 agggtttctc caaggacatc aaggtgccca agagccgcta cctgggctac atcaaggact
    10021 acgagggagc gacgctgatg gagtgtgagc tgaatccccg catcccctac acggagctgt
    10081 cccacatcat caagaagcag aaagagatca tcaagggatc cgattacaaa gacgatgacg
    10141 ataagggatc cggcgcaaca aacttctctc tgctgaaaca agccggagat gtcgaagaga
    10201 atcctggacc gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
    10261 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
    10321 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
    10381 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
    10441 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
    10501 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
    10561 ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc gcccgaccac cagggcaagg
    10621 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
    10681 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
    10741 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
    10801 gtgcctgaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
    10861 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
    10921 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
    10981 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
    11041 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
    11101 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
    11161 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
    11221 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
    11281 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
    11341 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
    11401 ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca
    11461 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
    11521 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
    11581 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
    11641 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
    11701 gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
    11761 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
    11821 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
    11881 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
    11941 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
    12001 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
    12061 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
    12121 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
    12181 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
    12241 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
    12301 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
    12361 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
    12421 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
    12481 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
    12541 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
    12601 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
    12661 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
    12721 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
    12781 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
    12841 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
    12901 accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg
    12961 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
    13021 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg
    13081 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg
    13141 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag
    13201 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg
    13261 gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa
    13321 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
    13381 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
    13441 taaagcaa
//
