LOCUS       sequence-256276-       11336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  targeted DNA demethylation_mouse_TSDR, expression of 
            dCas9-huTET1CD-T2A-mCherry and sgRNA6 targeting mouse TSDR .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11336)
  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 11336)
  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..11336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          94..321
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             342..368
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             372..392
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(402..425,426..428,429..2984,2985..2987,2988..4502)
                     /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(402..425,426..428,429..2915,2916..2918,2919..4502)
                     /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             6744..6797
                     /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             6798..7505
                     /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(6940..6958)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(7533..7550)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7539..7746
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7755..7895
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7755..7884
                     /label=AAV2 ITR
     rep_origin      7970..8425
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8057..8076)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8267..8288
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8442..8461)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8561..8583
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8621..8639)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8707..8811
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8812..8880,8881..9672)
                     /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(9030..9049)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9843..10431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10332..10351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        10493..10741
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     10493..10513
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     10664..10683
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        10762..10837
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        10934..11219
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
       61 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
      121 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      181 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
      241 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
      301 cctgaaatca ctttttttca ggttggaccg gtgccaccat gtacccatac gatgttccag
      361 attacgcttc gccgaagaaa aagcgcaagg tcgaagcgtc cgacaagaag tacagcatcg
      421 gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
      481 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
      541 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg
      601 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
      661 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
      721 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
      781 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
      841 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
      901 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
      961 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     1021 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     1081 agctgcccgg cgagaagaag aatggcctgt tcggcaacct gattgccctg agcctgggcc
     1141 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     1201 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     1261 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     1321 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     1381 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     1441 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca
     1501 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     1561 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     1621 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     1681 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     1741 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     1801 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg
     1861 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     1921 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga
     1981 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca
     2041 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     2101 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     2161 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     2221 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     2281 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     2341 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     2401 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     2461 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     2521 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     2581 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     2641 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     2701 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     2761 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     2821 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     2881 tggacatcaa ccggctgtcc gactacgatg tggacgccat cgtgcctcag agctttctga
     2941 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg
     3001 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga
     3061 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc
     3121 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     3181 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca
     3241 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     3301 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     3361 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     3421 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
     3481 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
     3541 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
     3601 aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga
     3661 gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
     3721 agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc
     3781 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca
     3841 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca
     3901 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
     3961 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
     4021 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc
     4081 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
     4141 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg
     4201 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
     4261 acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga
     4321 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
     4381 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga
     4441 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg
     4501 acaaaaggcc ggcggccacg aaaaaggccg gccagggagg aggatccctg cccacctgca
     4561 gctgtcttga tcgagttata caaaaagaca aaggcccata ttatacacac cttggggcag
     4621 gaccaagtgt tgctgctgtc agggaaatca tggagaatag gtatggtcaa aaaggaaacg
     4681 caataaggat agaaatagta gtgtacaccg gtaaagaagg gaaaagctct catgggtgtc
     4741 caattgctaa gtgggtttta agaagaagca gtgatgaaga aaaagttctt tgtttggtcc
     4801 ggcagcgtac aggccaccac tgtccaactg ctgtgatggt ggtgctcatc atggtgtggg
     4861 atggcatccc tcttccaatg gccgaccggc tatacacaga gctcacagag aatctaaagt
     4921 catacaatgg gcaccctacc gacagaagat gcaccctcaa tgaaaatcgt acctgtacat
     4981 gtcaaggaat tgatccagag acttgtggag cttcattctc ttttggctgt tcatggagta
     5041 tgtactttaa tggctgtaag tttggtagaa gcccaagccc cagaagattt agaattgatc
     5101 caagctctcc cttacatgaa aaaaaccttg aagataactt acagagtttg gctacacgat
     5161 tagctccaat ttataagcag tatgctccag tagcttacca aaatcaggtg gaatatgaaa
     5221 atgttgcccg agaatgtcgg cttggcagca aggaaggtcg acccttctct ggggtcactg
     5281 cttgcctgga cttctgtgct catccccaca gggacattca caacatgaat aatggaagca
     5341 ctgtggtttg taccttaact cgagaagata accgctcttt gggtgttatt cctcaagatg
     5401 agcagctcca tgtgctacct ctttataagc tttcagacac agatgagttt ggctccaagg
     5461 aaggaatgga agccaagatc aaatctgggg ccatcgaggt cctggcaccc cgccgcaaaa
     5521 aaagaacgtg tttcactcag cctgttcccc gttctggaaa gaagagggct gcgatgatga
     5581 cagaggttct tgcacataag ataagggcag tggaaaagaa acctattccc cgaatcaagc
     5641 ggaagaataa ctcaacaaca acaaacaaca gtaagccttc gtcactgcca accttaggga
     5701 gtaacactga gaccgtgcaa cctgaagtaa aaagtgaaac cgaaccccat tttatcttaa
     5761 aaagttcaga caacactaaa acttattcgc tgatgccatc cgctcctcac ccagtgaaag
     5821 aggcatctcc aggcttctcc tggtccccga agactgcttc agccacacca gctccactga
     5881 agaatgacgc aacagcctca tgcgggtttt cagaaagaag cagcactccc cactgtacga
     5941 tgccttcggg aagactcagt ggtgccaatg ctgcagctgc tgatggccct ggcatttcac
     6001 agcttggcga agtggctcct ctccccaccc tgtctgctcc tgtgatggag cccctcatta
     6061 attctgagcc ttccactggt gtgactgagc cgctaacgcc tcatcagcca aaccaccagc
     6121 cctccttcct cacctctcct caagaccttg cctcttctcc aatggaagaa gatgagcagc
     6181 attctgaagc agatgagcct ccatcagacg aacccctatc tgatgacccc ctgtcacctg
     6241 ctgaggagaa attgccccac attgatgagt attggtcaga cagtgagcac atctttttgg
     6301 atgcaaatat tggtggggtg gccatcgcac ctgctcacgg ctcggttttg attgagtgtg
     6361 cccggcgaga gctgcacgct accactcctg ttgagcaccc caaccgtaat catccaaccc
     6421 gcctctccct tgtcttttac cagcacaaaa acctaaataa gccccaacat ggttttgaac
     6481 taaacaagat taagtttgag gctaaagaag ctaagaataa gaaaatgaag gcctcagagc
     6541 aaaaagacca ggcagctaat gaaggtccag aacagtcctc tgaagtaaat gaattgaacc
     6601 aaattccttc tcataaagca ttaacattaa cccatgacaa tgttgtcacc gtgtcccctt
     6661 atgctctcac acacgttgcg gggccctata accattgggt catggccggc caggcaaaaa
     6721 agaaaaagga attcggcagt ggagagggca gaggaagtct gctaacatgc ggtgacgtcg
     6781 aggagaatcc tggcccagtg agcaagggcg aggaggataa catggccatc atcaaggagt
     6841 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     6901 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     6961 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     7021 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     7081 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     7141 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     7201 ccgacggccc cgtaatgcag aagaaaacca tgggctggga ggcctcctcc gagcggatgt
     7261 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     7321 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     7381 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     7441 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     7501 agtaagaatt ctaactagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     7561 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     7621 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     7681 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
     7741 tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     7801 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     7861 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     7921 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     7981 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     8041 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     8101 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     8161 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     8221 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     8281 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     8341 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     8401 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     8461 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     8521 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     8581 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     8641 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     8701 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     8761 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     8821 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     8881 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     8941 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     9001 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     9061 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     9121 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     9181 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     9241 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     9301 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     9361 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     9421 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     9481 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc
     9541 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     9601 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     9661 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     9721 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     9781 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     9841 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     9901 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     9961 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
    10021 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
    10081 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
    10141 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
    10201 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
    10261 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
    10321 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
    10381 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
    10441 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtgagggcct
    10501 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
    10561 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
    10621 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
    10681 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
    10741 cgtagccgga taggcccagg tgttttagag ctagaaatag caagttaaaa taaggctagt
    10801 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag agctagaaat
    10861 agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc agacaaatgg
    10921 ctctagaggt acccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
    10981 acccccgccc attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg
    11041 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
    11101 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc cagtacatga
    11161 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
    11221 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
    11281 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggg
//
