LOCUS       sequence-234970-       10135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor construct to amplify 
            HA-NLS-dCas9-NLS-SID4x-TaV-2a-Citrine-FRT-Kan-FRT cassette for BAC 
            recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10135)
  AUTHORS   Chong-Morrison V, Mayes S, Simoes FC, Senanayake U, Carroll DS, 
            Riley PR, Wilson SW, Sauka-Spengler T
  TITLE     Ac/Ds transposition for CRISPR/dCas9-SID4x epigenome modulation in 
            zebrafish.
  JOURNAL   Biol Open. 2023 Jun 15;12(6):bio059995. doi: 10.1242/bio.059995. 
            Epub 2023 Jun 27.
  PUBMED    37367831
REFERENCE   2  (bases 1 to 10135)
  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: "Biol 
            Open."; date: "2023-06-15"; volume: "12(6):bio059995. doi"; pages: "
            10.1242/bio.059995. Epub 2023 Jun 27"
COMMENT     Sequence Label: 
            pGEM-T-Easy-HA-NLS-dCas9-NLS-SID4x-TaV-2a-Citrine-FRT-Kan-FRT
FEATURES             Location/Qualifiers
     source          1..10135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..44
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     36..53
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     37..53
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     60..79
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        60..78
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      109..118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      113..122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             131..157
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             161..181
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(191..214,215..217,218..2773,2774..2776,2777..4291)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(191..214,215..217,218..2704,2705..2707,2708..4291)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4295..4315
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4736..4789
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4796..4798,4799..5509)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5099..5118)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5446..5467
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5564..5685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5575..5594)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5629..5648
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(5809..5856)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        5950..6054
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6056..6246
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6079..6099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             6286..7080
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6286..7080
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6340..6359)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6950..6969
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7087..7135
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    complement(7160..7207)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(7336..7354)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7337..7354)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7372..7388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7372..7388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7385..7407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7396..7412
                     /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(7420..7426,7427..7444,7445..7450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7465..7486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7603..7620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7774..8362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7854..7873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8533..9324,9325..9393))
                     /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     9156..9175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9394..9498)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9576..10031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9713..9734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9925..9944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt
       61 aatacgactc actatagggc gaattgggcc cgacgtcgca tgctcccggc cgccatggcg
      121 gccgcggatg tacccatacg atgttccaga ttacgcttcg ccgaagaaaa agcgcaaggt
      181 cgaagcgtcc gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg
      241 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
      301 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
      361 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
      421 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
      481 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
      541 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
      601 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
      661 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
      721 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
      781 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
      841 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
      901 cggcaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
      961 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     1021 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     1081 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     1141 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     1201 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     1261 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     1321 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     1381 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     1441 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     1501 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     1561 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     1621 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     1681 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     1741 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     1801 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     1861 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     1921 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     1981 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     2041 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     2101 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     2161 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     2221 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     2281 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     2341 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     2401 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     2461 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     2521 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     2581 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     2641 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     2701 ggacgccatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     2761 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     2821 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     2881 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     2941 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     3001 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     3061 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     3121 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     3181 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     3241 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     3301 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     3361 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     3421 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     3481 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     3541 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     3601 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     3661 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     3721 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     3781 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     3841 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     3901 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     3961 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     4021 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     4081 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     4141 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     4201 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     4261 acggatcgac ctgtctcagc tgggaggcga cagccccaag aagaagagaa aggtggaggc
     4321 cagcgggccg gccgaggcca gcatgaacat ccagatgctg ctggaggccg ctgactacct
     4381 ggaacggaga gagcgcgaag ccgagcacgg atatgcttca atgctgcccg gaagcggcat
     4441 gaatattcag atgctgctgg aggctgctga ttacctggaa aggcgcgaac gggaggccga
     4501 acatggctat gcttccatgc tgcctgggtc tggaatgaat atccaaatgc tgctggaggc
     4561 agccgattac ctggaacgga gagaaagaga agccgagcac ggatacgcca gcatgctgcc
     4621 aggcagcggg atgaacatac aaatgctgct ggaggctgcc gattacctgg agaggcgcga
     4681 gagagaagct gaacatggct atgcctctat gctgcccctc gagatcggat caggagaggg
     4741 caggggaagt cttctaacat gcggggacgt ggaggaaaat cccgggccca gatctgtgag
     4801 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4861 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     4921 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4981 caccttcggc tacggcctga tgtgcttcgc ccgctacccc gaccacatga agcagcacga
     5041 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaaaga
     5101 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5161 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5221 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     5281 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     5341 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5401 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5461 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
     5521 cgactctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     5581 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     5641 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     5701 attcatttta tgtttcgggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     5761 ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg acctgcttga agttcctata
     5821 ctttctagag aataggaact tcggaatagg aacttcaaga tccccctggc gaaaggggga
     5881 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cgtcaggtgg cacttttcgg
     5941 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6001 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     6061 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     6121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     6181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagtaacca
     6241 tagtccgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     6301 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6361 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6421 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     6481 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6541 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6601 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6661 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6721 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6781 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     6841 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6901 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6961 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     7021 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7081 ggggatcggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcggatc
     7141 cgagcttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag
     7201 taacttctcc atggtagcct ccaaaaaagc ctcctcacta cttctggaat agctcaatca
     7261 ctagtgaatt cgcggccgcc tgcaggtcga ccatatggga gagctcccaa cgcgttggat
     7321 gcatagcttg agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct
     7381 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7441 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7501 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7561 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7621 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7681 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7741 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7801 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7861 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7921 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7981 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     8041 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8101 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8161 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     8221 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8281 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8341 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8401 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8461 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8521 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8581 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8641 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8701 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8761 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8821 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8881 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8941 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     9001 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     9061 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     9121 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     9181 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9241 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     9301 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9361 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9421 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9481 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac
     9541 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     9601 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     9661 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     9721 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     9781 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     9841 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     9901 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     9961 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
    10021 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
    10081 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaa
//
