LOCUS       sequence-256197-       11370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inactivated control (targeted DNA demethylation_human_TSDR), 
            expression of dCas9-hudTET1CD-T2A-mCherry sgRNA3 targeting human 
            TSDR .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11370)
  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 11370)
  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-hudTET1CD-T2A-mCherry(PX458)-SghuTSDR-3
FEATURES             Location/Qualifiers
     source          1..11370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        155..403
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     155..175
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     326..345
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        424..499
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        596..881
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1126..1353
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1374..1400
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1404..1424
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1434..1457,1458..1460,1461..4016,4017..4019,
                     4020..5534)
                     /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(1434..1457,1458..1460,1461..3947,3948..3950,
                     3951..5534)
                     /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             7776..7829
                     /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             7830..8537
                     /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(7972..7990)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(8565..8582)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8571..8778
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8787..8927
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8787..8916
                     /label=AAV2 ITR
     rep_origin      9002..9457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9089..9108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9299..9320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9474..9493)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9593..9615
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9653..9671)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9739..9843
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9844..9912,9913..10704)
                     /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(10062..10081)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10875..11370,1..93)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(11364..11370,1..13)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
       61 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      121 ttcctggcct tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc
      181 atatttgcat atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa
      241 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
      301 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
      361 cgatttcttg gctttatata tcttgtggaa aggacgaaac accgggccag gtcttcagag
      421 ctagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
      481 agtggcaccg agtcggtgct tttttgtttt agagctagaa atagcaagtt aaaataaggc
      541 tagtccgttt ttagcgcgtg cgccaattct gcagacaaat ggctctagag gtacccgtta
      601 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      661 caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      721 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      781 atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta
      841 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
      901 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
      961 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggcggggcgg ggcgaggcgg
     1021 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
     1081 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
     1141 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     1201 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     1261 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
     1321 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atgtacccat
     1381 acgatgttcc agattacgct tcgccgaaga aaaagcgcaa ggtcgaagcg tccgacaaga
     1441 agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg
     1501 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     1561 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
     1621 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     1681 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     1741 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     1801 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     1861 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     1921 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     1981 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     2041 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     2101 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc
     2161 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     2221 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     2281 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     2341 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     2401 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     2461 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     2521 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     2581 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     2641 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     2701 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     2761 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     2821 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     2881 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     2941 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     3001 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     3061 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     3121 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     3181 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     3241 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     3301 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     3361 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     3421 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     3481 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     3541 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     3601 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     3661 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     3721 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     3781 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     3841 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     3901 tggaccagga actggacatc aaccggctgt ccgactacga tgtggacgcc atcgtgcctc
     3961 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
     4021 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     4081 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     4141 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     4201 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     4261 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     4321 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     4381 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     4441 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     4501 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     4561 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     4621 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     4681 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     4741 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
     4801 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     4861 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     4921 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     4981 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     5041 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     5101 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     5161 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
     5221 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     5281 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
     5341 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     5401 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     5461 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     5521 agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggccaggga ggaggatccc
     5581 tgcccacctg cagctgtctt gatcgagtta tacaaaaaga caaaggccca tattatacac
     5641 accttggggc aggaccaagt gttgctgctg tcagggaaat catggagaat aggtatggtc
     5701 aaaaaggaaa cgcaataagg atagaaatag tagtgtacac cggtaaagaa gggaaaagct
     5761 ctcatgggtg tccaattgct aagtgggttt taagaagaag cagtgatgaa gaaaaagttc
     5821 tttgtttggt ccggcagcgt acaggccacc actgtccaac tgctgtgatg gtggtgctca
     5881 tcatggtgtg ggatggcatc cctcttccaa tggccgaccg gctatacaca gagctcacag
     5941 agaatctaaa gtcatacaat gggcacccta ccgacagaag atgcaccctc aatgaaaatc
     6001 gtacctgtac atgtcaagga attgatccag agacttgtgg agcttcattc tcttttggct
     6061 gttcatggag tatgtacttt aatggctgta agtttggtag aagcccaagc cccagaagat
     6121 ttagaattga tccaagctct cccttacatg aaaaaaacct tgaagataac ttacagagtt
     6181 tggctacacg attagctcca atttataagc agtatgctcc agtagcttac caaaatcagg
     6241 tggaatatga aaatgttgcc cgagaatgtc ggcttggcag caaggaaggt cgacccttct
     6301 ctggggtcac tgcttgcctg gacttctgtg ctcatcccta cagggccatt cacaacatga
     6361 ataatggaag cactgtggtt tgtaccttaa ctcgagaaga taaccgctct ttgggtgtta
     6421 ttcctcaaga tgagcagctc catgtgctac ctctttataa gctttcagac acagatgagt
     6481 ttggctccaa ggaaggaatg gaagccaaga tcaaatctgg ggccatcgag gtcctggcac
     6541 cccgccgcaa aaaaagaacg tgtttcactc agcctgttcc ccgttctgga aagaagaggg
     6601 ctgcgatgat gacagaggtt cttgcacata agataagggc agtggaaaag aaacctattc
     6661 cccgaatcaa gcggaagaat aactcaacaa caacaaacaa cagtaagcct tcgtcactgc
     6721 caaccttagg gagtaacact gagaccgtgc aacctgaagt aaaaagtgaa accgaacccc
     6781 attttatctt aaaaagttca gacaacacta aaacttattc gctgatgcca tccgctcctc
     6841 acccagtgaa agaggcatct ccaggcttct cctggtcccc gaagactgct tcagccacac
     6901 cagctccact gaagaatgac gcaacagcct catgcgggtt ttcagaaaga agcagcactc
     6961 cccactgtac gatgccttcg ggaagactca gtggtgccaa tgctgcagct gctgatggcc
     7021 ctggcatttc acagcttggc gaagtggctc ctctccccac cctgtctgct cctgtgatgg
     7081 agcccctcat taattctgag ccttccactg gtgtgactga gccgctaacg cctcatcagc
     7141 caaaccacca gccctccttc ctcacctctc ctcaagacct tgcctcttct ccaatggaag
     7201 aagatgagca gcattctgaa gcagatgagc ctccatcaga cgaaccccta tctgatgacc
     7261 ccctgtcacc tgctgaggag aaattgcccc acattgatga gtattggtca gacagtgagc
     7321 acatcttttt ggatgcaaat attggtgggg tggccatcgc acctgctcac ggctcggttt
     7381 tgattgagtg tgcccggcga gagctgcacg ctaccactcc tgttgagcac cccaaccgta
     7441 atcatccaac ccgcctctcc cttgtctttt accagcacaa aaacctaaat aagccccaac
     7501 atggttttga actaaacaag attaagtttg aggctaaaga agctaagaat aagaaaatga
     7561 aggcctcaga gcaaaaagac caggcagcta atgaaggtcc agaacagtcc tctgaagtaa
     7621 atgaattgaa ccaaattcct tctcataaag cattaacatt aacccatgac aatgttgtca
     7681 ccgtgtcccc ttatgctctc acacacgttg cggggcccta taaccattgg gtcatggccg
     7741 gccaggcaaa aaagaaaaag gaattcggca gtggagaggg cagaggaagt ctgctaacat
     7801 gcggtgacgt cgaggagaat cctggcccag tgagcaaggg cgaggaggat aacatggcca
     7861 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     7921 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     7981 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     8041 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     8101 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     8161 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     8221 ccaacttccc ctccgacggc cccgtaatgc agaagaaaac catgggctgg gaggcctcct
     8281 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     8341 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     8401 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     8461 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     8521 acgagctgta caagtaagaa ttctaactag agctcgctga tcagcctcga ctgtgccttc
     8581 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     8641 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     8701 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa
     8761 tagcaggcat gctggggagc ggccgcagga acccctagtg atggagttgg ccactccctc
     8821 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     8881 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     8941 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     9001 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     9061 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     9121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     9181 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     9241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     9301 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     9361 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     9421 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     9481 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     9541 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     9601 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     9661 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     9721 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     9781 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     9841 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     9901 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     9961 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
    10021 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
    10081 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
    10141 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
    10201 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
    10261 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
    10321 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
    10381 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
    10441 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
    10501 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga
    10561 agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
    10621 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
    10681 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
    10741 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
    10801 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
    10861 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
    10921 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
    10981 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
    11041 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
    11101 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
    11161 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
    11221 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
    11281 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
    11341 gagcgcacga gggagcttcc agggggaaac
//
