LOCUS       sequence-307041-       16441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  OCT4, SOX2, KLF4, C-MYC, KLF2, NANOG (C6F) and tdTomato under the 
            control of TRE (Tet-ON).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16441)
  AUTHORS   Kisa F, Shiozawa S, Oda K, Yoshimatsu S, Nakamura M, Koya I, Kawai 
            K, Suzuki S, Okano H
  TITLE     Naive-like ESRRB(+) iPSCs with the Capacity for Rapid Neural 
            Differentiation.
  JOURNAL   Stem Cell Reports. 2017 Dec 12;9(6):1825-1838. doi: 
            10.1016/j.stemcr.2017.10.008. Epub 2017 Nov 9.
  PUBMED    29129686
REFERENCE   2  (bases 1 to 16441)
  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: 
            "10.1016/j.stemcr.2017.10.008"; journalName: "Stem Cell Reports"; 
            date: "2017-12-12- 12"; volume: "9"; issue: "6"; pages: "1825-1838"
COMMENT     Sequence Label: pPB-C6F/TdTomato
FEATURES             Location/Qualifiers
     source          1..16441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   660..694
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     1097..1113
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1120..1138
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1238..1359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1303..1322
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1496..2926)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     regulatory      2923..2932
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2946..3498
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2948..3521
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             complement(4546..4611)
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     regulatory      5697..5706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        5760..6147
                     /label=bidirectional TRE promoter
                     /note="Tet-responsive bidirectional promoter PTight-BI, 
                     consisting of seven tet operator sequences flanked on each 
                     side by the minimal CMV promoter"
     protein_bind    5834..5852
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5870..5888
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5905..5923
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5941..5959
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5977..5995
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6012..6030
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6048..6066
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        6166..6184
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             7314..7379
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             8826..8891
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     misc_feature    9884..10457
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             10502..11821
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     misc_feature    join(11848..12422,12423..12425,12426..12434)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             12435..13034
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(12435..12454)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     12931..12951
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             13176..13409
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(13549..13566)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13555..13779
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(13826..13842)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    13850..13866
                     /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(13874..13880,13881..13898,13899..13904))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13919..13940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   14167..14229
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(14253..14271)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(14292..14308)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    14316..14332
                     /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(14340..14346,14347..14364,14365..14370))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14385..14406
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14523..14540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14694..15282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14774..14793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15453..16244,16245..16313))
                     /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     16076..16095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16314..16418)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccttaac
      661 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
      721 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
      781 cgtgaggcgt gcttgtcaat gaggtaagtg tcactgattt tgaactataa cgaccgcgtg
      841 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
      901 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
      961 tagatgtata ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     1021 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     1081 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
     1141 gaattgggcc ctctagatgc atgctcgacg gtatacagac atgataagat acattgatga
     1201 gtttggacaa accacaacta gaatgcactc gacgcgttaa gatacattga tgagtttgga
     1261 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     1321 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1381 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     1441 aaatgtggta tggctgatta tgatcagtta tctagatgca tgctcgagcg gccgcctact
     1501 tgtacagctc gtccatgccg tacaggaaca ggtggtggcg gccctcggag cgctcgtact
     1561 gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc cagcttggtg tccacgtagt
     1621 agtagccggg cagttgcacg ggcttcttgg ccatgtagat ggtcttgaac tccaccaggt
     1681 agtggccgcc gtccttcagc ttcagggcct ggtggatctc gcccttcagc acgccgtcgc
     1741 gggggtacag gcgctcggtg gaggcctccc agcccatggt cttcttctgc attacggggc
     1801 cgtcgggggg gaagttggtg ccgcgcatct tcaccttgta gatcagcgtg ccgtcctgca
     1861 gggaggagtc ctgggtcacg gtcaccagac cgccgtcctc gaagttcatc acgcgctccc
     1921 acttgaagcc ctcggggaag gacagcttct tgtaatcggg gatgtcggcg gggtgcttca
     1981 cgtacgcctt ggagccgtac atgaactggg gggacaggat gtcccaggcg aagggcaggg
     2041 ggccgccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc
     2101 cctcgccctc gatctcgaac tcgtggccgt tcatggagcc ctccatgcgc accttgaagc
     2161 gcatgaactc tttgatgacg gccatgttgt tgtcctcgga ggaggcggtg ccggagctgc
     2221 cgctgccggt gctgccggtg ccatgcccca ggaacaggtg gtggcggccc tcggagcgct
     2281 cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc ttggtgtcca
     2341 cgtagtagta gccgggcagt tgcacgggct tcttggccat gtagatggtc ttgaactcca
     2401 ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg gatctcgccc ttcagcacgc
     2461 cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc catggtcttc ttctgcatta
     2521 cggggccgtc gggggggaag ttggtgccgc gcatcttcac cttgtagatc agcgtgccgt
     2581 cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc gtcctcgaag ttcatcacgc
     2641 gctcccactt gaagccctcg gggaaggaca gcttcttgta atcggggatg tcggcggggt
     2701 gcttcacgta cgccttggag ccgtacatga actgggggga caggatgtcc caggcgaagg
     2761 gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc
     2821 cctcgccctc gccctcgatc tcgaactcgt ggccgttcat ggagccctcc atgcgcacct
     2881 tgaagcgcat gaactctttg atgacctcct cgcccttgct caccatggtg gcggatccgt
     2941 ggccatatta tcatcgtgtt tttcaaagga aaaccacgtc cccgtggttc ggggggccta
     3001 gacgtttttt taacctcgac taaacacatg taaagcatgt gtaccgaggc cccagatcag
     3061 atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat acgcttgagg
     3121 agagccattt gactctttcc acaactatcc aactcacaac gtggcactgg ggttgtgccg
     3181 cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg tcgccaggtg
     3241 gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa gaagcttcca
     3301 gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg agaggggaaa
     3361 gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc acattgccaa
     3421 aagacggcaa tatggtggaa aataacatat agacaaacgc acaccggcct tattccaagc
     3481 ggcttcggcc agtaacgtta gggggggggg agggagaggg gcggactgtg gaaaaaaaag
     3541 ggagatccgc tagaagggcg aattcgaatc cggagacgta cgaccggtta acaggcctta
     3601 agcgctagat gcatgctcga ctcacacgtc ttcaggttgc atgttcatgg agtagtttag
     3661 gaataaatcc atggtttgtg gagtactaaa atacctagtg gtctgctgta ttacattaag
     3721 gccttcccca gcagcttcca aagcagcctc caagtcactg gcaggagaat ttggctggaa
     3781 ctgcatgcag gactgcagag attcctctcc acagttatag aagggactgt tccaggcctg
     3841 attgttccag gattgggtgc accaggtctg agtgttccag gagtggttgc tccaggactg
     3901 gatgttctgg gtctggttgc tccaggttga attgttccag gtctggttgc tccacattgg
     3961 aaggttccca gtcgggttca ccaggcatcc ctggtggtag gaagagtaga ggctggggta
     4021 ggtaggtgct gaggccttct gcgtcacacc attgctattc ttcggccagt tgtttttctg
     4081 ccacctctta gatttcattc tctggttctg gaaccaggtc ttcacctgtt tgtagctgag
     4141 gttcaggatg ttggagagtt cttgcatctg ctggaggctg aggtatttct gtctctgaaa
     4201 tctatcattg agtacacaca gctgggtgga agagaacaca gttctggtct tctgtttctt
     4261 gactgggacc ttgtcttcct tttttgcgac actattctct gcagaagtgg gttgtttgcc
     4321 tttgggactg gtggaagaat cagggctgtc ctgaataagc agatccatgg aggagggaag
     4381 aggagagaca gtctccgtgt gaggcatctc agcagaagac atttgcaagg atggatagtt
     4441 ttcttcaggc ccacaaatca caggcatagg tgaagattct ttacagtcgg atgcttcaaa
     4501 gcaaggcaag ctttggggac aagctggatc cacactcatg atatccggcc ctgggttgga
     4561 ctccacgtct cccgccaact tgagaaggtc aaaattcaaa gtctgtttca ccggtgcccg
     4621 cttagcgcga ggatccatgt gccgtttcat gtgcagcgcc aggtgatcgg agcgcgagaa
     4681 ggcacgatcg cacagatggc actggaatgg ccggtggccc gtgtgctttc ggtagtggcg
     4741 cgtgagctcg tctgagcgcg caaacttcca gccgcagccg tcccagttgc agtggtaggg
     4801 cttctcacct gtgtgcgtgc gcagatgcgc cttcagatgc gaactcttgg tgtaggtctt
     4861 gccgcagccc gcgtagctgc aggtgtgagt ggcggtgcgt ttgcggggcc aagagcggcg
     4921 gccgcgcttt ggcttggcct ccagcagctc cagcggggac gcaggcggcg tgaggagacc
     4981 gcgggcggcg gggggcgcca ggcccagggc tgccgcggcg gcggccgcgt cgtcgaagag
     5041 accgaaggct gggggcgcag gcggcgcgta atgcaggccg ggcccgggcg cgccgaaacc
     5101 agggccaccg aaaggcggcg ggaaggaggc gcgcggaccg ggcgcgggca ggcgcgcggg
     5161 gccgtcgggg ctgagcggcg gtgtgtcggg cggcggcggg gggcggcccc cgggacctcg
     5221 catgcacgaa gccgccgggc ccggggcgcc ctcgcgcttg aggccgcgcg gtccacgggt
     5281 cagcccgggg gcgcagccgt agccgccgcc gccgtccgct tcagggggct cggccttgac
     5341 caggcggccg ggcggcgcca gcagaaagcg gccgtgcagt gcgggccccg gcggcgcatc
     5401 cagctcgggt cgcagcagct cagacaccag gccacccgcg ggggcgctgt agggcggggg
     5461 cgcgccgggt tcggggtaat agaacgcagg cggcgggggc ggcggcggcg gctccggggc
     5521 ggcctcggcg cccaggccat ccagccccat ggacaggatg aagtccagca cgctgttgag
     5581 gtcgtcgtcg gtgccgccgg actcgggttc ggcgcgcggc cagcgctcct gcaggccgcg
     5641 ctcgcggcac gggctggcga aagtggagaa ggacggcagg atgggttcac tcagcgccat
     5701 ggtggctagc acgcgtcagc tgactagagg atccccgggt accgagctcg aaatctccag
     5761 gcgatctgac ggttcactaa acgagctctg cttatatagg cctcccaccg tacacgccta
     5821 cctcgacata cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact
     5881 gatagggata aactcgacat acgttctcta tcactgatag ggagtaaact cgacatacgt
     5941 tctctatcac tgatagggag taaactcgac atacgttctc tatcactgat agggagtaaa
     6001 ctcgacatcg ttctctatca ctgataggga gtaaactcga catacgttct ctatcactga
     6061 tagggagtaa actcgagtat gtcgaggtgg cgtgtacggt gggaggccta tataagcaga
     6121 gctcgtttag tgaaccgtca gatcgcctgg agaattcgcc taggcaatta accctcacta
     6181 aagggaaggc cgcctgggcc acgcgtgcca tggcgggaca cctggcttcg gatttcgcct
     6241 tctcgccccc tccaggtggt ggaggtgatg ggccaggggg gccggagccg ggctgggttg
     6301 atcctcggac ctggctaagc ttccaaggcc ctcctggagg gccaggaatc gggccggggg
     6361 ttgggccagg ctctgaggtg tgggggattc ccccatgccc cccgccgtat gagttctgtg
     6421 gggggatggc gtactgtggg ccccaggttg gagtggggct agtgccccaa ggcggcttgg
     6481 agacctctca gcctgagggt gaagcaggag tcggggtgga gagcaactcc gatggggcct
     6541 ccccggagcc ctgcaccgtc acccctggtg ccgtgaagct ggagaaggag aagctggagc
     6601 aaaacccgga ggagtcccag gacatcaaag ctctgcagaa agaactcgag caatttgcca
     6661 agctcctgaa gcagaagagg atcaccctgg gatatacaca ggccgatgtg gggctcaccc
     6721 tgggggttct atttgggaag gtattcagcc aaacgaccat ctgccgcttt gaggctctgc
     6781 agcttagctt caagaacatg tgtaagctgc ggcccttgct gcagaagtgg gtggaggaag
     6841 ctgacaacaa tgaaaatctt caggagatat gcaaagcaga aaccctcgtg caggcccgaa
     6901 agagaaagcg aaccagtatc gagaaccgag tgagaggcaa cctggagaat ttgttcctgc
     6961 agtgcccgaa acccacactg cagcagatca gccacatcgc ccagcagctt gggctcgaga
     7021 aggatgtggt ccgagtgtgg ttctgtaacc ggcgccagaa gggcaagcga tcaagcagcg
     7081 actatgcaca acgagaggat tttgaggctg ctgggtctcc tttctcaggg ggaccagtgt
     7141 cctttcctct ggccccaggg ccccattttg gtaccccagg ctatgggagc cctcacttca
     7201 ctgcactgta ctcctcggtc cctttccctg agggggaagc ctttccccct gtctctgtca
     7261 ccactctggg ctctcccatg cattcaaacg gggcgcgtaa aattgtcgct cctgtcaaac
     7321 aaactcttaa ctttgattta ctcaaactgg ctggggatgt agaaagcaat ccaggtccag
     7381 gggcgcgtgg tatggctgtc agcgacgcgc tgctcccatc tttctccacg ttcgcgtctg
     7441 gcccggcggg aagggagaag acactgcgtc aagcaggtgc cccgaataac cgctggcggg
     7501 aggagctctc ccacatgaag cgacttcccc cagtgcttcc cggccgcccc tatgacctgg
     7561 cggcggcgac cgtggccaca gacctggaga gcggcggagc cggtgcggct tgcggcggta
     7621 gcaacctggc gcccctacct cggagagaga ccgaggagtt caacgatctc ctggacctgg
     7681 actttattct ctccaattcg ctgacccatc ctccggagtc agtggccgcc accgtgtcct
     7741 cgtcagcgtc agcctcctct tcgtcgtcgc cgtcgagcag cggccctgcc agcgcgccct
     7801 ccacctgcag cttcacctat ccgatccggg ccgggaacga cccgggcgtg gcgccgggcg
     7861 gcacgggcgg aggcctcctc tatggcaggg agtccgctcc ccctccgacg gctcccttca
     7921 acctggcgga catcaacgac gtgagcccct cgggcggctt cgtggccgag ctcctgcggc
     7981 cagaattgga cccggtgtac attccgccgc agcagccgca gccgccaggt ggcgggctga
     8041 tgggcaagtt cgtgctgaag gcgtcgctga gcgcccctgg cagcgagtac ggcagcccgt
     8101 cggtcatcag cgtcagcaaa ggcagccctg acggcagcca cccggtggtg gtggcgccct
     8161 acaacggcgg gccgccgcgc acgtgcccca agatcaagca ggaggcggtc tcttcgtgca
     8221 cccacttggg cgctggaccc cctctcagca atggccaccg gccggctgca cacgacttcc
     8281 ccctggggcg gcagctcccc agcaggacta ccccgaccct gggtcttgag gaagtgctga
     8341 gcagcaggga ctgtcaccct gccctgccgc ttcctcccgg cttccatccc cacccggggc
     8401 ccaattaccc atccttcctg cccgatcaga tgcagccgca agtcccgccg ctccattacc
     8461 aagagctcat gccacccggt tcctgcatgc cagaggagcc caagccaaag aggggaagac
     8521 gatcgtggcc ccggaaaagg accgccaccc acacttgtga ttacgcgggc tgcggcaaaa
     8581 cctacacaaa gagttcccat ctcaaggcac acctgcgaac ccacacaggt gagaaacctt
     8641 accactgtga ctgggacggc tgtggatgga aattcgcccg ctcagatgaa ctgaccaggc
     8701 actaccgtaa acacacgggg caccgcccgt tccagtgcca aaaatgcgac cgagcatttt
     8761 ccaggtcgga ccacctcgcc ttacacatga agaggcattt tggggcgcgt aaaattgtcg
     8821 ctcctgtcaa acaaactctt aactttgatt tactcaaact ggctggggat gtagaaagca
     8881 atccaggtcc aggggcgcgt ggtatgtaca acatgatgga gacggagctg aagccgccgg
     8941 gcccgcagca aacttcgggg ggcggcggcg gcaactccac cgcggcggcg gccggcggca
     9001 accagaaaaa cagcccggac cgcgtcaagc ggcccatgaa tgccttcatg gtgtggtccc
     9061 gcgggcagcg gcgcaagatg gcccaggaga accccaagat gcacaactcg gagatcagca
     9121 agcgcctggg cgccgagtgg aaacttttgt cggagacgga gaagcggccg ttcatcgacg
     9181 aggctaagcg gctgcgagcg ctgcacatga aggagcaccc ggattataaa taccggcccc
     9241 ggcggaaaac caagacgctc atgaagaagg ataagtacac gctgcccggc gggctgctgg
     9301 cccccggcgg caatagcatg gcgagcgggg tcggggtggg cgccggcctg ggcgcgggcg
     9361 tgaaccagcg catggacagt tacgcgcaca tgaacggctg gagcaacggc agctacagca
     9421 tgatgcagga ccagctgggc tacccgcagc acccgggcct caatgcgcac ggcgcagcgc
     9481 agatgcagcc catgcaccgc tacgacgtga gcgccctgca gtacaactcc atgaccagct
     9541 cgcagaccta catgaacggc tcgcccacct acagcatgtc ctactcgcag cagggcaccc
     9601 ctggcatggc tcttggctcc atgggttcgg tggtcaagtc cgaggccagc tccagccccc
     9661 ctgtggttac ctcttcctcc cactccaggg cgccctgcca ggccggggac ctccgggaca
     9721 tgatcagcat gtatctcccc ggcgccgagg tgccggaacc cgccgccccc agcagacttc
     9781 acatgtccca gcactaccag agcggcccgg tgcccggcac ggccattaac ggcacactgc
     9841 ccctctcaca catgtgagtt taaatctagg gcggccaatt ccgcccctct cccccccccc
     9901 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     9961 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
    10021 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
    10081 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
    10141 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
    10201 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
    10261 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
    10321 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
    10381 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
    10441 gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca tggcgcgtgg
    10501 tatgcccctc aacgttagct tcaccaacag gaactatgac ctcgactacg actcggtgca
    10561 gccgtatttc tactgcgacg aggaggagaa cttctaccag cagcagcagc agagcgagct
    10621 gcagcccccg gcgcccagcg aggatatctg gaagaaattc gagctgctgc ccaccccgcc
    10681 cctgtcccct agccgccgct ccgggctctg ctcgccctcc tacgttgcgg tcacaccctt
    10741 ctcccttcgg ggagacaacg acggcggtgg cgggagcttc tccacggccg accagctgga
    10801 gatggtgacc gagctgctgg gaggagacat ggtgaaccag agtttcatct gcgacccgga
    10861 cgacgagacc ttcatcaaaa acatcatcat ccaggactgt atgtggagcg gcttctcggc
    10921 cgccgccaag ctcgtctcag agaagctggc ctcctaccag gctgcgcgca aagacagcgg
    10981 cagcccgaac cccgcccgcg gccacagcgt ctgctccacc tccagcttgt acctgcagga
    11041 tctgagcgcc gccgcctcag agtgcatcga cccctcggtg gtcttcccct accctctcaa
    11101 cgacagcagc tcgcccaagt cctgcgcctc gcaagactcc agcgccttct ctccgtcctc
    11161 ggattctctg ctctcctcga cggagtcctc cccgcagggc agccccgagc ccctggtgct
    11221 ccatgaggag acaccgccca ccaccagcag cgactctgag gaggaacaag aagatgagga
    11281 agaaatcgat gttgtttctg tggaaaagag gcaggctcct ggcaaaaggt cagagtctgg
    11341 atcaccttct gctggaggcc acagcaaacc tcctcacagc ccactggtcc tcaagaggtg
    11401 ccacgtctcc acacatcagc acaactacgc agcgcctccc tccactcgga aggactatcc
    11461 tgctgccaag agggtcaagt tggacagtgt cagagtcctg agacagatca gcaacaaccg
    11521 aaaatgcacc agccccaggt cctcggacac cgaggagaat gtcaagaggc gaacacacaa
    11581 cgtcttggag cgccagagga ggaacgagct aaaacggagc ttttttgccc tgcgtgacca
    11641 gatcccggag ttggaaaaca atgaaaaggc ccccaaggta gttatcctta aaaaagccac
    11701 agcatacatc ctgtccgtcc aagcagagga gcaaaagctc atttctgaag aggacttgtt
    11761 gcggaaacga cgagaacagt tgaaacacaa acttgaacag ctacggaact cttgtgcgta
    11821 agtttatctg cagaattccg cccccccccc ctctccctcc ccccccccta acgttactgg
    11881 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
    11941 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
    12001 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
    12061 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
    12121 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
    12181 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
    12241 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
    12301 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
    12361 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
    12421 atatggccac aaccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
    12481 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
    12541 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
    12601 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
    12661 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
    12721 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
    12781 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
    12841 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
    12901 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
    12961 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
    13021 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
    13081 cccatgcatc gatttcgaac ccggggtacc tttaagacca atgacttaca aggcagctgt
    13141 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaaag
    13201 aagacaagat ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
    13261 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
    13321 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
    13381 cttttagtca gtgtggaaaa tctctagcag aattccagca cactggcggc cgttactagt
    13441 ggatcatcat aatttaaaca agcaaaacca aattaagggc cagctcattc ctcccactca
    13501 tgatctatag atcctaagct tatcgatgat ctagagctcg ctgatcagcc tcgactgtgc
    13561 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
    13621 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
    13681 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
    13741 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacct
    13801 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
    13861 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
    13921 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
    13981 tgtcgtgcca gcggatcctt tgttacttta tagaagaaat tttgagtttt tgtttttttt
    14041 taataaataa ataaacataa ataaattgtt tgttgaattt attattagta tgtaagtgta
    14101 aatataataa aacttaatat ctattcaaat taataaataa acctcgatat acagaccgat
    14161 aaaacacatg cgtcaatttt acgcatgatt atctttaacg tacgtcacaa tatgattatc
    14221 tttctagggt taaagagctc cagcttttgt tccctttagt gagggttaat ttcgagcttg
    14281 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    14341 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    14401 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    14461 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
    14521 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    14581 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    14641 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    14701 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    14761 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    14821 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    14881 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    14941 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    15001 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    15061 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    15121 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    15181 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    15241 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
    15301 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
    15361 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
    15421 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
    15481 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
    15541 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
    15601 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
    15661 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
    15721 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
    15781 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
    15841 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
    15901 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    15961 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    16021 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    16081 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    16141 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    16201 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacgggaagg
    16261 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    16321 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    16381 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    16441 c
//
