LOCUS       sequence-210074-        8471 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  To knock-out TDP43.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8471)
  AUTHORS   Schmidt HB, Barreau A, Rohatgi R
  TITLE     Phase separation-deficient TDP43 remains functional in splicing.
  JOURNAL   Nat Commun. 2019 Oct 25;10(1):4890. doi: 10.1038/s41467-019-12740-2.
  PUBMED    31653829
REFERENCE   2  (bases 1 to 8471)
  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: "Nat 
            Commun."; date: "2019-10-25"; volume: "10(1):4890. doi"; pages: " 
            10.1038/s41467-019-12740-2"
COMMENT     Sequence Label: pHBS952 TARDBP-Exon1-ko-sgRNA1-SpCas9
FEATURES             Location/Qualifiers
     source          1..8471
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      2..11
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             11..76
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             83..103
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             128..4228
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             128..4228
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4229..4276
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4292..4345
                     /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             4346..5050
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(4488..4506)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(5078..5095)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5084..5291
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5300..5440
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5300..5429
                     /label=AAV2 ITR
     rep_origin      5515..5970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5602..5621)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5812..5833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5987..6006)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6106..6128
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6166..6184)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6252..6356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6357..6425,6426..7217)
                     /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     complement(6575..6594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7388..7976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7877..7896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8038..8286
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8038..8058
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8209..8228
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8308..8383
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tgccaccatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
       61 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
      121 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
      181 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
      241 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
      301 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
      361 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
      421 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
      481 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
      541 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
      601 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
      661 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
      721 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
      781 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
      841 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
      901 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
      961 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     1021 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     1081 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     1141 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     1201 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     1261 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     1321 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     1381 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     1441 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     1501 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     1561 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     1621 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     1681 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     1741 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     1801 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     1861 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     1921 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     1981 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     2041 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     2101 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     2161 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     2221 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     2281 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     2341 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     2401 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     2461 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     2521 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     2581 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     2641 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     2701 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     2761 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     2821 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     2881 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     2941 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     3001 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     3061 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     3121 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     3181 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     3241 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     3301 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     3361 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     3421 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     3481 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     3541 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     3601 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     3661 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     3721 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     3781 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     3841 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     3901 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     3961 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     4021 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     4081 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     4141 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     4201 gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca
     4261 ggcaaaaaag aaaaaggaat tcggcagtgg agagggcaga ggaagtctgc taacatgcgg
     4321 tgacgtcgag gagaatcctg gcccagtgag caagggcgag gaggataaca tggccatcat
     4381 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     4441 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     4501 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     4561 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     4621 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     4681 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     4741 cttcccctcc gacggccccg taatgcagaa gaaaaccatg ggctgggagg cctcctccga
     4801 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     4861 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     4921 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     4981 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     5041 gctgtacaag gaattctaac tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     5101 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     5161 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     5221 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg
     5281 catgctgggg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     5341 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     5401 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     5461 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     5521 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5581 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5641 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5701 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5761 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5821 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     5881 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5941 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     6001 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     6061 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     6121 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     6181 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6241 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6301 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6361 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6421 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6481 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6541 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6601 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6661 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6721 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6781 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6841 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6901 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6961 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7021 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg
     7081 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7141 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7201 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7261 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7321 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7381 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7441 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7501 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7561 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7621 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7681 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7741 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7801 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7861 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7921 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7981 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtgag
     8041 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     8101 attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     8161 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     8221 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     8281 aacaccgaga tgctggctgg ggaaatcgtt ttagagctag aaatagcaag ttaaaataag
     8341 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
     8401 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
     8461 aaatggctct a
//
