LOCUS       sequence-256270-       11352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  targeted DNA demethylation_mouse_TSDR, expression of 
            dCas9-huTET1CD-T2A-mCherry and sgRNA8 targeting mouse TSDR .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11352)
  AUTHORS   Kressler C, Gasparoni G, Nordstrom K, Hamo D, Salhab A, 
            Dimitropoulos C, Tierling S, Reinke P, Volk HD, Walter J, Hamann A, 
            Polansky JK
  TITLE     Targeted De-Methylation of the FOXP3-TSDR Is Sufficient to Induce 
            Physiological FOXP3 Expression but Not a Functional Treg Phenotype.
  JOURNAL   Front Immunol. 2021 Jan 7;11:609891. doi: 10.3389/fimmu.2020.609891.
            eCollection 2020.
  PUBMED    33488615
REFERENCE   2  (bases 1 to 11352)
  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: "Front 
            Immunol."; date: "2021-01-7"; volume: "11:609891. doi"; pages: " 
            10.3389/fimmu.2020.609891. eCollection 2020"
COMMENT     Sequence Label: pSpdCas9-huTET1CD-T2A-mCherry(PX458)-SgmouseTS...
FEATURES             Location/Qualifiers
     source          1..11352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          110..337
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             358..384
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             388..408
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(418..441,442..444,445..3000,3001..3003,3004..4518)
                     /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(418..441,442..444,445..2931,2932..2934,2935..4518)
                     /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             6760..6813
                     /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             6814..7521
                     /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(6956..6974)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(7549..7566)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7555..7762
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7771..7911
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7771..7900
                     /label=AAV2 ITR
     rep_origin      7986..8441
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8073..8092)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8283..8304
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8458..8477)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8577..8599
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8637..8655)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8723..8827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8828..8896,8897..9688)
                     /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(9046..9065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9859..10447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10348..10367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        10509..10757
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     10509..10529
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     10680..10699
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        10778..10853
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        10950..11235
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
       61 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      121 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      181 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      241 ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa
      301 ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc caccatgtac
      361 ccatacgatg ttccagatta cgcttcgccg aagaaaaagc gcaaggtcga agcgtccgac
      421 aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc
      481 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
      541 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
      601 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
      661 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
      721 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
      781 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
      841 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
      901 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
      961 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     1021 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     1081 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg caacctgatt
     1141 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     1201 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     1261 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     1321 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     1381 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     1441 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     1501 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     1561 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     1621 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     1681 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     1741 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     1801 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     1861 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     1921 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     1981 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     2041 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     2101 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     2161 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     2221 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     2281 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     2341 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     2401 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     2461 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     2521 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     2581 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     2641 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     2701 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     2761 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     2821 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     2881 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga cgccatcgtg
     2941 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     3001 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     3061 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     3121 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     3181 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     3241 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     3301 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     3361 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     3421 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     3481 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     3541 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     3601 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     3661 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     3721 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     3781 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     3841 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     3901 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     3961 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     4021 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     4081 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     4141 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     4201 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     4261 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     4321 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     4381 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     4441 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     4501 tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca gggaggagga
     4561 tccctgccca cctgcagctg tcttgatcga gttatacaaa aagacaaagg cccatattat
     4621 acacaccttg gggcaggacc aagtgttgct gctgtcaggg aaatcatgga gaataggtat
     4681 ggtcaaaaag gaaacgcaat aaggatagaa atagtagtgt acaccggtaa agaagggaaa
     4741 agctctcatg ggtgtccaat tgctaagtgg gttttaagaa gaagcagtga tgaagaaaaa
     4801 gttctttgtt tggtccggca gcgtacaggc caccactgtc caactgctgt gatggtggtg
     4861 ctcatcatgg tgtgggatgg catccctctt ccaatggccg accggctata cacagagctc
     4921 acagagaatc taaagtcata caatgggcac cctaccgaca gaagatgcac cctcaatgaa
     4981 aatcgtacct gtacatgtca aggaattgat ccagagactt gtggagcttc attctctttt
     5041 ggctgttcat ggagtatgta ctttaatggc tgtaagtttg gtagaagccc aagccccaga
     5101 agatttagaa ttgatccaag ctctccctta catgaaaaaa accttgaaga taacttacag
     5161 agtttggcta cacgattagc tccaatttat aagcagtatg ctccagtagc ttaccaaaat
     5221 caggtggaat atgaaaatgt tgcccgagaa tgtcggcttg gcagcaagga aggtcgaccc
     5281 ttctctgggg tcactgcttg cctggacttc tgtgctcatc cccacaggga cattcacaac
     5341 atgaataatg gaagcactgt ggtttgtacc ttaactcgag aagataaccg ctctttgggt
     5401 gttattcctc aagatgagca gctccatgtg ctacctcttt ataagctttc agacacagat
     5461 gagtttggct ccaaggaagg aatggaagcc aagatcaaat ctggggccat cgaggtcctg
     5521 gcaccccgcc gcaaaaaaag aacgtgtttc actcagcctg ttccccgttc tggaaagaag
     5581 agggctgcga tgatgacaga ggttcttgca cataagataa gggcagtgga aaagaaacct
     5641 attccccgaa tcaagcggaa gaataactca acaacaacaa acaacagtaa gccttcgtca
     5701 ctgccaacct tagggagtaa cactgagacc gtgcaacctg aagtaaaaag tgaaaccgaa
     5761 ccccatttta tcttaaaaag ttcagacaac actaaaactt attcgctgat gccatccgct
     5821 cctcacccag tgaaagaggc atctccaggc ttctcctggt ccccgaagac tgcttcagcc
     5881 acaccagctc cactgaagaa tgacgcaaca gcctcatgcg ggttttcaga aagaagcagc
     5941 actccccact gtacgatgcc ttcgggaaga ctcagtggtg ccaatgctgc agctgctgat
     6001 ggccctggca tttcacagct tggcgaagtg gctcctctcc ccaccctgtc tgctcctgtg
     6061 atggagcccc tcattaattc tgagccttcc actggtgtga ctgagccgct aacgcctcat
     6121 cagccaaacc accagccctc cttcctcacc tctcctcaag accttgcctc ttctccaatg
     6181 gaagaagatg agcagcattc tgaagcagat gagcctccat cagacgaacc cctatctgat
     6241 gaccccctgt cacctgctga ggagaaattg ccccacattg atgagtattg gtcagacagt
     6301 gagcacatct ttttggatgc aaatattggt ggggtggcca tcgcacctgc tcacggctcg
     6361 gttttgattg agtgtgcccg gcgagagctg cacgctacca ctcctgttga gcaccccaac
     6421 cgtaatcatc caacccgcct ctcccttgtc ttttaccagc acaaaaacct aaataagccc
     6481 caacatggtt ttgaactaaa caagattaag tttgaggcta aagaagctaa gaataagaaa
     6541 atgaaggcct cagagcaaaa agaccaggca gctaatgaag gtccagaaca gtcctctgaa
     6601 gtaaatgaat tgaaccaaat tccttctcat aaagcattaa cattaaccca tgacaatgtt
     6661 gtcaccgtgt ccccttatgc tctcacacac gttgcggggc cctataacca ttgggtcatg
     6721 gccggccagg caaaaaagaa aaaggaattc ggcagtggag agggcagagg aagtctgcta
     6781 acatgcggtg acgtcgagga gaatcctggc ccagtgagca agggcgagga ggataacatg
     6841 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     6901 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     6961 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     7021 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     7081 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     7141 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     7201 ggcaccaact tcccctccga cggccccgta atgcagaaga aaaccatggg ctgggaggcc
     7261 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     7321 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     7381 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     7441 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     7501 atggacgagc tgtacaagta agaattctaa ctagagctcg ctgatcagcc tcgactgtgc
     7561 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     7621 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     7681 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     7741 agaatagcag gcatgctggg gagcggccgc aggaacccct agtgatggag ttggccactc
     7801 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     7861 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     7921 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     7981 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     8041 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     8101 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     8161 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     8221 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     8281 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     8341 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     8401 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     8461 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     8521 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     8581 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     8641 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     8701 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     8761 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     8821 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     8881 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     8941 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     9001 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     9061 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     9121 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     9181 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     9241 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     9301 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     9361 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     9421 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     9481 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     9541 tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     9601 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     9661 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     9721 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     9781 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     9841 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     9901 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     9961 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
    10021 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
    10081 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
    10141 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
    10201 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
    10261 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
    10321 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
    10381 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
    10441 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
    10501 tcacatgtga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
    10561 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
    10621 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
    10681 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
    10741 ggaaaggacg aaacaccggg cttcatcggc aacaagggtt ttagagctag aaatagcaag
    10801 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
    10861 ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa
    10921 ttctgcagac aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct
    10981 ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc
    11041 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
    11101 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
    11161 tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
    11221 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
    11281 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
    11341 gcgggggggg gg
//
