LOCUS       sequence-367392-       10296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Cas9 nuclease and mStrawberry via T2A linker under 
            control of a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10296)
  AUTHORS   Abatti LE, Lado-Fernandez P, Huynh L, Collado M, Hoffman MM, 
            Mitchell JA
  TITLE     Epigenetic reprogramming of a distal developmental enhancer cluster 
            drives SOX2 overexpression in breast and lung adenocarcinoma.
  JOURNAL   Nucleic Acids Res. 2023 Sep 22:gkad734. doi: 10.1093/nar/gkad734.
  PUBMED    37738673
REFERENCE   2  (bases 1 to 10296)
  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: "Nucleic 
            Acids Res. 2023 Sep 22:gkad734. doi: 10.1093/nar/gkad734."
COMMENT     Sequence Label: CMV-hCas9-T2A-mStrawberry
FEATURES             Location/Qualifiers
     source          1..10296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        190..569
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        570..773
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     724..744
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     770..794
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      869..878
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             875..4978
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             875..4978
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             875..4978
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4991..5011
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5018..5071
                     /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             5078..5785
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mStrawberry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILTPNFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKMRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVELYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(5220..5238)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5488..5507
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5822..5841)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5866..5914
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6116..6544
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6203..6222)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6413..6434
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6553..6573)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6558..6887
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6738..6873
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6800..6819
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6954..7748
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7008..7027)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7618..7637
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7924..8045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7961..7980)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8015..8034
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8094..8110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8094..8110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8107..8129)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8118..8134
                     /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(8142..8148,8149..8166,8167..8172))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8187..8208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8325..8342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8496..9084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8576..8595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9255..10046,10047..10115))
                     /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     9878..9897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10116..10220)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(10295..10296,1..18))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
       61 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
      121 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
      181 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      241 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      301 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      361 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      421 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      481 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      541 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      601 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      661 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      721 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      781 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      841 gggaccgatc cagcctccgg actctagagc caccatggac aagaagtact ccattgggct
      901 cgatatcggc acaaacagcg tcggctgggc cgtcattacg gacgagtaca aggtgccgag
      961 caaaaaattc aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg
     1021 cgccctcctg ttcgactccg gggagacggc cgaagccacg cggctcaaaa gaacagcacg
     1081 gcgcagatat acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga
     1141 gatggctaag gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga
     1201 ggataaaaag cacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca
     1261 tgaaaagtac ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc
     1321 tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct
     1381 catcgagggg gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt
     1441 tcagacttac aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa
     1501 agcaatcctg agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct
     1561 ccctggggag aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac
     1621 ccccaacttt aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga
     1681 cacctacgat gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct
     1741 ttttttggcg gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa
     1801 cacggagatc accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca
     1861 ccaagacttg actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga
     1921 aattttcttc gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca
     1981 ggaggaattt tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct
     2041 gctggtaaag cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag
     2101 catcccccac cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt
     2161 ctaccccttt ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc
     2221 ctactatgta ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc
     2281 agaagagacc atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca
     2341 gtccttcatc gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc
     2401 taaacactct ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata
     2461 cgtcacagaa gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt
     2521 ggacctcctc ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagactattt
     2581 caaaaagatt gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc
     2641 atccctggga acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa
     2701 tgaggagaac gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag
     2761 ggagatgatt gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa
     2821 acagctcaag aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg
     2881 gatccgagac aagcagagtg gaaagacaat cctggatttt cttaagtccg atggatttgc
     2941 caaccggaac ttcatgcagt tgatccatga tgactctctc acctttaagg aggacatcca
     3001 gaaagcacaa gtttctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg
     3061 tagcccagct atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa
     3121 agtaatggga aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac
     3181 tacccagaag ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa
     3241 agaactgggg tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga
     3301 gaagctctac ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga
     3361 catcaatcgg ctctccgact acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga
     3421 tgattctatt gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa
     3481 cgtcccctca gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc
     3541 caaactgatc acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc
     3601 tgagttggat aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa
     3661 gcacgtggcc caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact
     3721 gattcgagag gtgaaagtta ttactctgaa gtctaagctg gtctcagatt tcagaaagga
     3781 ctttcagttt tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct
     3841 gaatgcagtg gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt
     3901 ttacggagac tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat
     3961 aggcaaggcc accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga
     4021 gattacactg gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac
     4081 aggagaaatc gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat
     4141 gccgcaggtg aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag
     4201 tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa
     4261 gaaatacggc ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt
     4321 ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat
     4381 ggagcgatca agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga
     4441 ggtcaaaaaa gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg
     4501 ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc
     4561 ctctaaatac gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc
     4621 cgaagataat gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat
     4681 catcgagcaa ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa
     4741 ggtgctttct gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat
     4801 tatccacttg tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac
     4861 caccatagac agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca
     4921 tcagtcaatt acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag
     4981 cagggctgac cccaagaaga agaggaaggt ggctagcgag ggcagaggaa gtcttctaac
     5041 atgcggtgac gtggaggaga atcccggccc tggtaccgtg agcaagggcg aggagaataa
     5101 catggccatc atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
     5161 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     5221 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc taacccccaa
     5281 cttcacctac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     5341 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     5401 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     5461 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     5521 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagatgag
     5581 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     5641 gaagcccgtg cagctgcccg gcgcctacat cgtcggcatc aagttggaca tcacctccca
     5701 caacgaggac tacaccatcg tggaactgta cgaacgcgcc gagggccgcc actccaccgg
     5761 cggcatggac gagctgtaca agtaaaccgg ttagtaatga gtttaaacgg gggaggctaa
     5821 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5881 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     5941 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     6001 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     6061 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     6121 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6181 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6241 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6301 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6361 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6421 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6481 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6541 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     6601 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6661 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     6721 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6781 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     6841 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     6901 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     6961 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     7021 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     7081 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     7141 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     7201 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     7261 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     7321 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     7381 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     7441 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     7501 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     7561 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     7621 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     7681 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     7741 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     7801 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     7861 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     7921 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7981 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     8041 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     8101 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     8161 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     8221 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     8281 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     8341 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     8401 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     8461 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     8521 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8581 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     8641 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     8701 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     8761 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8821 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     8881 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     8941 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     9001 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     9061 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     9121 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     9181 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     9241 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     9301 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     9361 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     9421 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     9481 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     9541 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     9601 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     9661 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     9721 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     9781 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     9841 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     9901 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     9961 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
    10021 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
    10081 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
    10141 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
    10201 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
    10261 ggagatctcc cgatccccta tggtgcactc tcagta
//
