LOCUS       sequence-334608-       10467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To introduce dCas9-fused Tet1-CD and sgRNA2.0 system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10467)
  AUTHORS   Xu X, Tao Y, Gao X, Zhang L, Li X, Zou W, Ruan K, Wang F, Xu GL, Hu 
            R
  TITLE     A CRISPR-based approach for targeted DNA demethylation.
  JOURNAL   Cell Discov. 2016 May 3;2:16009. doi: 10.1038/celldisc.2016.9. 
            eCollection 2016.
  PUBMED    27462456
REFERENCE   2  (bases 1 to 10467)
  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: "Cell 
            Discov."; date: "2016-05-3"; volume: "2:16009. doi"; pages: " 
            10.1038/celldisc.2016.9. eCollection 2016"
COMMENT     Sequence Label: pdCas9-Tet1-CD
FEATURES             Location/Qualifiers
     source          1..10467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        285..303
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        355..373
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     enhancer        416..701
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        703..980
                     /label=chicken beta-actin promoter
     intron          981..1208
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1229..1255
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1259..1279
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1289..1312,1313..1315,1316..3802,3803..3805,
                     3806..5389)
                     /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             1289..5389
                     /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="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             5393..5413
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7508..7525)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7514..7721
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7730..7870
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7730..7859
                     /label=AAV2 ITR
     rep_origin      7945..8400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8032..8051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8242..8263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8417..8436)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8536..8558
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8596..8614)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8682..8786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8787..8855,8856..9647)
                     /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(9005..9024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9818..10406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10307..10326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag gccaacatga ggatcaccca
      301 tgtctgcagg gcctagcaag ttaaaataag gctagtccgt tatcaacttg gccaacatga
      361 ggatcaccca tgtctgcagg gccaagtggc accgagtcgg tgcttttttg gtacccgtta
      421 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      481 caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      541 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      601 atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta
      661 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
      721 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
      781 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccrggsgggg
      841 sggggsgggg sgrggggsgg ggsggggsgr ggcggagagg tgcggcggca gccaatcaga
      901 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
      961 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
     1021 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     1081 ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa gggtttaagg
     1141 gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct gaaatcactt
     1201 tttttcaggt tggaccggtg ccaccatgta cccatacgat gttccagatt acgcttcgcc
     1261 gaagaaaaag cgcaaggtcg aagcgtccga caagaagtac agcatcggcc tggccatcgg
     1321 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt
     1381 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct
     1441 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata
     1501 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa
     1561 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa
     1621 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
     1681 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg
     1741 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg
     1801 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
     1861 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct
     1921 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga
     1981 gaagaagaat ggcctgttcg gcaacctgat tgccctgagc ctgggcctga cccccaactt
     2041 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga
     2101 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc
     2161 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat
     2221 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct
     2281 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt
     2341 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt
     2401 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa
     2461 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca
     2521 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt
     2581 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt
     2641 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac
     2701 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat
     2761 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag
     2821 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga
     2881 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct
     2941 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat
     3001 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg
     3061 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa
     3121 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat
     3181 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa
     3241 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga
     3301 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa
     3361 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca
     3421 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc
     3481 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg
     3541 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa
     3601 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg
     3661 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta
     3721 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg
     3781 gctgtccgac tacgatgtgg acgctatcgt gcctcagagc tttctgaagg acgactccat
     3841 cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc
     3901 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat
     3961 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga
     4021 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc
     4081 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga
     4141 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt
     4201 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
     4261 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
     4321 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
     4381 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
     4441 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
     4501 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
     4561 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
     4621 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
     4681 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
     4741 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
     4801 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
     4861 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
     4921 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
     4981 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
     5041 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
     5101 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
     5161 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
     5221 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
     5281 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
     5341 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca gccccaagaa
     5401 gaagagaaag gtggaggcca gcggtggagg aagtggaggt ggttcaggtg gaggatccgg
     5461 cgaagctgca ccctgtgact gtgatggagg tacacaaaaa gaaaaaggcc catattatac
     5521 acaccttggg gcaggaccaa gtgtggctgc tgtcagggag ctcatggaga ctaggtttgg
     5581 ccagaagggg aaggcaatcc ggattgagaa gatagtgttc acggggaagg aagggaagag
     5641 ctctcagggc tgcccggtcg ccaagtgggt gatcagaaga agtggtcctg aagagaagct
     5701 tatttgtttg gttcgtgagc gtgtagacca tcactgttcg acggctgtga tagttgtcct
     5761 tatcctgctg tgggaaggta tccctcgcct gatggctgac cgcctgtaca aagagctcac
     5821 tgagaacttg aggtcctaca gcggacatcc cacagaccga agatgtaccc tcaacaaaaa
     5881 gcgtacctgc acctgtcaag gcatcgaccc aaaaacctgc ggagcgtcct tctcctttgg
     5941 ctgttcgtgg agcatgtatt tcaacggctg taagtttggg aggagtgaaa accccagaaa
     6001 attcagactt gctccaaact accccttaca taactactat aagagaatta ctggaatgag
     6061 ttctgaagga agtgacgtga aaaccgggtg gatcattcca gaccgcaaga ccctcataag
     6121 cagagaggaa aaacagcttg aaaagaattt acaagaattg gctacagtat tagctccact
     6181 ttacaagcag atggctccag ttgcttatca aaatcaggtg gaatatgaag aagttgctgg
     6241 agactgtcga cttggaaatg aagaggggcg tcctttctct ggtgtcacct gttgcatgga
     6301 tttttgtgcc cattctcaca aggacattca caacatgcac aacggaagca ccgtggtgtg
     6361 tacgttgatt cgagcagatg gccgtgacac aaattgtccc gaggatgaac aactccacgt
     6421 cctgccacta taccggcttg cagacactga tgaatttggc tccgtggaag ggatgaaggc
     6481 caaaatcaaa tctggggcca tccaagtcaa tgggccaacc aggaagaggc gactacgttt
     6541 tactgagcct gttcctcgat gtgggaagag ggccaaaatg aagcagaacc acaataaatc
     6601 aggttcacac aacactaaga gcttttcatc agcctcatct acttctcacc tagtgaaaga
     6661 cgaatctaca gacttctgtc ccctgcaggc ttcctccgca gaaacatcta cctgtacgta
     6721 cagtaaaaca gcctcaggtg ggtttgcaga aacaagtagt attctccact gcacaatgcc
     6781 ttctggagca cacagtggtg ctaatgcagc tgctggggaa tgtactggaa cggtgcagcc
     6841 tgccgaggtg gctgctcatc ctcaccagtc tcttcccaca gccgattctc ccgttcatgc
     6901 tgagcctctc actagtccat ctgagcagct aacttctaac cagtcaaacc agcagctccc
     6961 tctcctcagc aattctcaga aactggcttc ctgtcaggtg gaagatgagc ggcaccctga
     7021 agcggatgag cctcagcacc ccgaggacga taacttgcct caacttgatg agttctggtc
     7081 agacagtgag gagatctacg ccgatccttc ctttggtggc gtggcgatag cacccattca
     7141 cggctcggtg ctcattgagt gcgctcggaa ggagcttcat gctaccacct ctttgcgctc
     7201 ccccaaacga ggggtccctt ttcgtgtgtc ccttgtattc taccagcaca aaagcctaaa
     7261 caagcctaat catggttttg atatcaacaa aattaagtgt aaatgcaaaa aagtaacgaa
     7321 aaaaaagccc gcagaccggg agtgtcctga tgtatccccc gaagccaatt tatcacacca
     7381 aattccttct cgagttgcat caaccttaac ccgagacaat gttgttaccg tgtccccata
     7441 ctctctcact catgttgcgg gaccctacaa tcgttgggtc taagaattcc tagagctcgc
     7501 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     7561 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     7621 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     7681 aagggggagg attgggaaga gaatagcagg catgctgggg agcggccgca ggaaccccta
     7741 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     7801 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     7861 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     7921 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     7981 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     8041 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     8101 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     8161 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     8221 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     8281 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     8341 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     8401 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     8461 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     8521 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     8581 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     8641 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     8701 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     8761 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     8821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     8881 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     8941 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     9001 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     9061 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     9121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     9181 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     9241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     9301 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     9361 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     9421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     9481 aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta
     9541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     9601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     9661 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     9721 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     9781 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     9841 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     9901 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     9961 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
    10021 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
    10081 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
    10141 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
    10201 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
    10261 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
    10321 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
    10381 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
    10441 ccttttgctg gccttttgct cacatgt
//
