LOCUS       sequence-319753-       12875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 oscillator v1 (dCRv1) + [dCas9 + PA4-mVenus]. (Integration of 
            413; pOSIP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12875)
  AUTHORS   Kuo J, Yuan R, Sanchez C, Paulsson J, Silver PA
  TITLE     Toward a translationally independent RNA-based synthetic oscillator 
            using deactivated CRISPR-Cas.
  JOURNAL   Nucleic Acids Res. 2020 Aug 20;48(14):8165-8177. doi: 
            10.1093/nar/gkaa557.
  PUBMED    32609820
REFERENCE   2  (bases 1 to 12875)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2020-08-20- 20"; volume: "48"; issue: 
            "14"; pages: "8165-8177"
COMMENT     Sequence Label: pJK423
FEATURES             Location/Qualifiers
     source          1..12875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        128..177
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             207..218
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             227..940
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             227..940
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(257..285)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     880..900
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1001..1087
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1100..1194)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1215..1874)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1789..1808
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1875..1977)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(2007..2633)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     2538..2557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        2649..2704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    2685..2703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2722..2733
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2740..2766,2767..2769,2770..5256,5257..5259,
                     5260..6846)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     terminator      6870..6941
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6957..6984
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        complement(7065..7140)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(7161..7195)
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     misc_RNA        complement(7479..7554)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      7637..7680
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7702..7722
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      7777..7804
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(7853..7884)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    7961..7994
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(8001..8714)
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             8803..8811
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     RBS             8823..8845
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             8853..9863
                     /codon_start=1
                     /gene="int"
                     /product="integrase from phage 186"
                     /label=phage 186 integrase
                     /translation="MTVRKNPAGGWICELYPNGAKGKRIRKKFATKGEALAFEQYTVQN
                     PWQEEKEDRRTLKGLVDSWYSAHGITLKDGLKRQLAMHHAFECMGEPLARDFDAQMFSR
                     YREKRLKGEYARSNRVKEVSPRTLNLELAYFRAVFNELNRLGEWKGENPLKNMRPFRTE
                     EMEMTWLTHDQISQLLGECNRHDHPDLETVVRICLATGARWSEAESLRKSQLAKYKITY
                     TNTKGRKNRTVPISKELYESLPDDKKGRLFSDCYGAFRSALERTGIELPAGQLTHVLRH
                     TFASHFMMNGGNILVLQRVLGHTDIKMTMRYAHFAPDHLEDAVKLNPLVHITNSK"
     primer_bind     complement(10026..10045)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             10037..10852
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(10110..10129)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      10974..11362
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    11373..11420
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    11433..11647
                     /gene="attP"
                     /label=phage 186 attP
                     /bound_moiety="phage 186 integrase"
                     /note="attachment site of phage 186"
     terminator      complement(11764..12010)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     primer_bind     complement(12101..12123)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     terminator      12333..12419
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      12516..12602
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      12699..12785
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gaattcgagc tcggtacccg gggatcctct agagggctta ctaaaagcca gataacagta
       61 tgcgtatttg cgcgcctagt atgcgtctcc tttacacctc cacaactagc tggtattatg
      121 ctagcagctg tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct
      181 ctacaaataa ttttgtttaa ctttacaaag aggagaaagg taccgcatga gtaaaggaga
      241 agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
      301 caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa
      361 attgatttgc actactggaa aactacctgt tccatggcca acacttgtca ctactttggg
      421 ttatggtgtt caatgctttg cgagataccc agatcatatg aaacagcatg actttttcaa
      481 gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa
      541 ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
      601 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
      661 taactcacac aatgtataca tcacggcaga caaacaaaag aatggaatca aagcgaactt
      721 caaaattaga cacaacattg aagatggagg tgttcaacta gcagaccatt atcaacaaaa
      781 tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt cctaccaatc
      841 taagctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac
      901 agctgctggg attacacatg gcatggatga actatacaaa taagcttgat atcgaattcc
      961 tgcagcccgg gggatcccat tgtacgcgtg ctagaggcat caaataaaac gaaaggctca
     1021 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     1081 gacaaattct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     1141 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     1201 agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     1261 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     1321 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     1381 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     1441 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     1501 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     1561 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     1621 catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg
     1681 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     1741 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     1801 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     1861 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     1921 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg
     1981 tctcattttc gccagatatc gacgtcttaa gacccacttt cacatttaag ttgtttttct
     2041 aatccgcata tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca
     2101 aataattcga tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt
     2161 tcttctttag cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc
     2221 acagcgctga gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct
     2281 aattgatttt cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt
     2341 gctccatcgc gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa
     2401 tcttgccagc tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca
     2461 atggctaagg cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc
     2521 tctacaccta gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta
     2581 agcagctcta atgcgctgtt aatcacttta cttttatcta atctagacat cattaattcc
     2641 taatttttgt tgacactcta tcgttgatag agttatttta ccactcccta tcagtgatag
     2701 agaaaagaat tcaaaagatc taaagaggag aaaggatcta tggataagaa atactcaata
     2761 ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt
     2821 ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt
     2881 ataggggctc ttttatttga cagtggagag acagcggaag cgactcgtct caaacggaca
     2941 gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca
     3001 aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg
     3061 gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct
     3121 tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat
     3181 aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat
     3241 tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag
     3301 ttggtacaaa cctacaatca attatttgaa gaaaacccta ttaacgcaag tggagtagat
     3361 gctaaagcga ttctttctgc acgattgagt aaatcaagac gattagaaaa tctcattgct
     3421 cagctccccg gtgagaagaa aaatggctta tttgggaatc tcattgcttt gtcattgggt
     3481 ttgaccccta attttaaatc aaattttgat ttggcagaag atgctaaatt acagctttca
     3541 aaagatactt acgatgatga tttagataat ttattggcgc aaattggaga tcaatatgct
     3601 gatttgtttt tggcagctaa gaatttatca gatgctattt tactttcaga tatcctaaga
     3661 gtaaatactg aaataactaa ggctccccta tcagcttcaa tgattaaacg ctacgatgaa
     3721 catcatcaag acttgactct tttaaaagct ttagttcgac aacaacttcc agaaaagtat
     3781 aaagaaatct tttttgatca atcaaaaaac ggatatgcag gttatattga tgggggagct
     3841 agccaagaag aattttataa atttatcaaa ccaattttag aaaaaatgga tggtactgag
     3901 gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac ctttgacaac
     3961 ggctctattc cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa
     4021 gacttttatc catttttaaa agacaatcgt gagaagattg aaaaaatctt gacttttcga
     4081 attccttatt atgttggtcc attggcgcgt ggcaatagtc gttttgcatg gatgactcgg
     4141 aagtctgaag aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca
     4201 gctcaatcat ttattgaacg catgacaaac tttgataaaa atcttccaaa tgaaaaagta
     4261 ctaccaaaac atagtttgct ttatgagtat tttacggttt ataacgaatt gacaaaggtc
     4321 aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc
     4381 attgttgatt tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat
     4441 tatttcaaaa aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt
     4501 aatgcttcat taggtaccta ccatgatttg ctaaaaatta ttaaagataa agattttttg
     4561 gataatgaag aaaatgaaga tatcttagag gatattgttt taacattgac cttatttgaa
     4621 gatagggaga tgattgagga aagacttaaa acatatgctc acctctttga tgataaggtg
     4681 atgaaacagc ttaaacgtcg ccgttatact ggttggggac gtttgtctcg aaaattgatt
     4741 aatggtatta gggataagca atctggcaaa acaatattag attttttgaa atcagatggt
     4801 tttgccaatc gcaattttat gcagctgatc catgatgata gtttgacatt taaagaagac
     4861 attcaaaaag cacaagtgtc tggacaaggc gatagtttac atgaacatat tgcaaattta
     4921 gctggtagcc ctgctattaa aaaaggtatt ttacagactg taaaagttgt tgatgaattg
     4981 gtcaaagtaa tggggcggca taagccagaa aatatcgtta ttgaaatggc acgtgaaaat
     5041 cagacaactc aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt
     5101 atcaaagaat taggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa
     5161 aatgaaaagc tctatctcta ttatctccaa aatggaagag acatgtatgt ggaccaagaa
     5221 ttagatatta atcgtttaag tgattatgat gtcgatgcca ttgttccaca aagtttcctt
     5281 aaagacgatt caatagacaa taaggtctta acgcgttctg ataaaaatcg tggtaaatcg
     5341 gataacgttc caagtgaaga agtagtcaaa aagatgaaaa actattggag acaacttcta
     5401 aacgccaagt taatcactca acgtaagttt gataatttaa cgaaagctga acgtggaggt
     5461 ttgagtgaac ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgccaaatc
     5521 actaagcatg tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat
     5581 aaacttattc gagaggttaa agtgattacc ttaaaatcta aattagtttc tgacttccga
     5641 aaagatttcc aattctataa agtacgtgag attaacaatt accatcatgc ccatgatgcg
     5701 tatctaaatg ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaatcggag
     5761 tttgtctatg gtgattataa agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa
     5821 gaaataggca aagcaaccgc aaaatatttc ttttactcta atatcatgaa cttcttcaaa
     5881 acagaaatta cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga aactaatggg
     5941 gaaactggag aaattgtctg ggataaaggg cgagattttg ccacagtgcg caaagtattg
     6001 tccatgcccc aagtcaatat tgtcaagaaa acagaagtac agacaggcgg attctccaag
     6061 gagtcaattt taccaaaaag aaattcggac aagcttattg ctcgtaaaaa agactgggat
     6121 ccaaaaaaat atggtggttt tgatagtcca acggtagctt attcagtcct agtggttgct
     6181 aaggtggaaa aagggaaatc gaagaagtta aaatccgtta aagagttact agggatcaca
     6241 attatggaaa gaagttcctt tgaaaaaaat ccgattgact ttttagaagc taaaggatat
     6301 aaggaagtta aaaaagactt aatcattaaa ctacctaaat atagtctttt tgagttagaa
     6361 aacggtcgta aacggatgct ggctagtgcc ggagaattac aaaaaggaaa tgagctggct
     6421 ctgccaagca aatatgtgaa ttttttatat ttagctagtc attatgaaaa gttgaagggt
     6481 agtccagaag ataacgaaca aaaacaattg tttgtggagc agcataagca ttatttagat
     6541 gagattattg agcaaatcag tgaattttct aagcgtgtta ttttagcaga tgccaattta
     6601 gataaagttc ttagtgcata taacaaacat agagacaaac caatacgtga acaagcagaa
     6661 aatattattc atttatttac gttgacgaat cttggagctc ccgctgcttt taaatatttt
     6721 gatacaacaa ttgatcgtaa acgatatacg tctacaaaag aagttttaga tgccactctt
     6781 atccatcaat ccatcactgg tctttatgaa acacgcattg atttgagtca gctaggaggt
     6841 gactaactcg agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact
     6901 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     6961 caccttcggg tgggcctttc tgcgtttata cctagataaa ggaccaaaac gaaaaaagac
     7021 gcttttcagc gtcttattgt tcgtctttgg taccgagaaa aaaagcaccg actcggtgcc
     7081 actttttcaa gttgataacg gactagcctt attttaactt gctatttcta gctctaaaac
     7141 acgggtcaca cgggtattat gctagcataa tacctaggac tgagctaggt gtaaaggatc
     7201 ctatcggacc aaaacgaaaa aagacgctcg aaagcgtctc ttttctggaa tttggtaccg
     7261 agaaaaaaag caccgactcg gtgccacttt ttcaagttgc aatcggacta gccttatttt
     7321 aacttgctat ttctagctct aaaacagctc agtcctaggt attatgctag cataatacca
     7381 gctagttgtg gaggtgtaaa aagcttaagc ggaccaaaac gaaaaaaggc ccccctttcg
     7441 ggaggcctct tttctggaat ttggtaccga gaaaaaaagc accgactcgg tgccactttt
     7501 tcaagttgat aacggactag ccttatttta acttgctatt tctagctcta aaacccacaa
     7561 ctagctggta ttatgctagc ataatacccg tgtgacccgt tggtgtaaaa tgatttctga
     7621 gatctttaga attccagaaa tcatccttag cgaaagctaa ggattttttt tatctgaaat
     7681 tctgcctcgt gatacgccta ttactagtag cggccgctgc aggcatgcct cgagatgcat
     7741 ggcgcctaac ctaaactgac aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     7801 tttttgcgtt tcgaacaatt gaaaaaacct cgcgccttac ctgttgagta atagtcaaaa
     7861 gcctccggtc ggaggctttt gactttctgc ttactgaatt tcggtggtgc cgttaattaa
     7921 ccggtgggcc ctcatgataa taatggtttc ttagacgtcc gaagttccta ttctctagaa
     7981 agtataggaa cttccctagg tcagccaaac gtctcttcag gccactgact agcgataact
     8041 ttccccacaa cggaacaact ctcattgcat gggatcattg ggtactgtgg gtttagtggt
     8101 tgtaaaaaca cctgaccgct atccctgatc agtttcttga aggtaaactc atcaccccca
     8161 agtctggcta tgcagaaatc acctggctca acagcctgct cagggtcaac gagaattaac
     8221 attccgtcag gaaagcttgg cttggagcct gttggtgcgg tcatggaatt accttcaacc
     8281 tcaagccaga atgcagaatc actggctttt ttggttgtgc ttacccatct ctccgcatca
     8341 cctttggtaa aggttctaag cttaggtgag aacatccctg cctgaacatg agaaaaaaca
     8401 gggtactcat actcacttct aagtgacggc tgcatactaa ccgcttcata catctcgtag
     8461 atttctctgg cgattgaagg gctaaattct tcaacgctaa ctttgagaat ttttgtaagc
     8521 aatgcggcgt tataagcatt taatgcattg atgccattaa ataaagcacc aacgcctgac
     8581 tgccccatcc ccatcttgtc tgcgacagat tcctgggata agccaagttc atttttcttt
     8641 ttttcataaa ttgctttaag gcgacgtgcg tcctcaagct gctcttgtgt taatggtttc
     8701 ttttttgtgc tcatacgtta aatctatcac cgcaagggat aaatatctaa caccgtgcgt
     8761 gttgactatt ttacctctgg cggtgataat ggttgcatgt actaaggagg ttgtatacgc
     8821 gttttgttta actttaagaa ggagatatac atatgaccgt ccgtaaaaat ccggctggcg
     8881 gttggatttg tgaactctac ccaaacggtg caaaaggcaa acgtatcaga aagaaattcg
     8941 ctactaaagg cgaggctctg gcgtttgagc agtacaccgt tcaaaacccg tggcaggaag
     9001 aaaaggaaga caggcgcacg ttaaaagggc tggttgattc atggtatagc gctcatggca
     9061 ttacactgaa agatggtttg aaacgccagt tagccatgca ccatgctttt gagtgtatgg
     9121 gcgaaccact cgcacgcgat ttcgatgcgc agatgttttc ccgctaccga gaaaaacggt
     9181 taaaaggtga gtatgcccgt tcaaacagag tgaaagaggt atcgcctcgc acgcttaatc
     9241 ttgagctggc ctacttccgg gcggtgttca atgagctaaa ccgcctcgga gaatggaagg
     9301 gtgaaaaccc actaaaaaat atgcgcccat tccgcacaga agaaatggaa atgacttggc
     9361 taactcacga ccaaatttcg caactgctcg gagagtgtaa ccgacatgac caccctgatt
     9421 tagaaaccgt ggtaagaatc tgtctcgcaa ctggcgcacg gtggtctgag gccgagagtc
     9481 tgagaaaaag ccaactcgcg aaatacaaaa tcacatacac caacacgaaa ggtagaaaaa
     9541 atcgcaccgt cccaatcagc aaagagctct atgagtctct gcctgatgat aaaaagggcc
     9601 ggttgttcag tgattgttat ggcgcgttcc ggtcagcttt ggaaagaaca ggtatcgaac
     9661 taccggcagg acaacttacc cacgttttac gtcacacctt cgctagtcac tttatgatga
     9721 atggcggtaa tattttggtg ttgcagcgcg tactcggcca tacagatata aaaatgacca
     9781 tgcgatatgc gcactttgca cccgaccatt tagaggatgc agttaaacta aatcctttag
     9841 ttcacattac taacagcaaa taagttatgg aggttgtatg tcagttcggc tagcccatgg
     9901 gtatggacag ttttcccttt gatatgtaac gcacgttgtg tctcaaaatc tctgatgtta
     9961 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
    10021 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctccc gtccgcgctt
    10081 aaactccaac atggacgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
    10141 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
    10201 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtccgtc tcaactggct
    10261 gacggagttt atgcctctcc cgaccatcaa gcattttatc cgtactcctg atgatgcgtg
    10321 gttactcacc accgcgattc ctgggaaaac agccttccag gtattagaag aatatcctga
    10381 ttcaggtgaa aatattgttg atgcgctggc cgtgttcctg cgccggttac attcgattcc
    10441 tgtttgtaat tgtcctttta acagcgatcg tgtatttcgt cttgctcagg cgcaatcacg
    10501 catgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
    10561 tgaacaagtc tggaaagaaa tgcacaagct cttgccattc tcaccggatt cagtcgtcac
    10621 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
    10681 tgatgttgga cgggtcggaa tcgcagaccg ttaccaggac cttgccattc tttggaactg
    10741 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
    10801 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aataatacta
    10861 gcagaaatca tccttagcga aagctaagga ttttttttat ctgattaccg cctttgagtg
    10921 agcgtcgacc tagtgcggcc gcaagatccg gccacgatgc gtccggcgta gaggatctga
    10981 agatcagcag ttcaacctgt tgatagtacg tactaagctc tcatgtttca cgtactaagc
    11041 tctcatgttt aacgtactaa gctctcatgt ttaacgaact aaaccctcat ggctaacgta
    11101 ctaagctctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
    11161 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
    11221 agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca
    11281 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca
    11341 caggaacact taacggctga catgggaatt aggaagttcc tattccgaag ttcctattct
    11401 ctagaaagta taggaacttc catatgccat gggcgatggt tctgagtaac agataataga
    11461 atggcgataa agtggcggta gaaatggcga ataataggta attattggca aacaatggca
    11521 atctatgtca atgataaata acgcatacta ttgattttcg gttgttcttg taggaactca
    11581 taatcgcttg gtcgctggtt caagtccagc aggggccacc aaatttaagt agtaaataca
    11641 tacagttagg ccactctctc gagtggcctt tttgttgcct cgttttcgag tggcgataga
    11701 atggcggtgg rttttttacc gccactttcc tgaggagtgc atagctagct tcttcgtctg
    11761 tttctactgg tattggcaca aacctgattc caatttgagc aaggctatgt gccatctcga
    11821 tactcgttct taactcaaca gaagatgctt tgtgcataca gcccctcgtt tattatttat
    11881 ctcctcagcc agccgctgtg ctttcagtgg atttcggata acagaaaggc cgggaaatac
    11941 ccagcctcgc tttgtaacgg agtagagacg aaagtgattg cgcctacccg gatattatcg
    12001 tgaggatgcg tcatcgccat taattcactg atcagtgata agctgtcaaa catgagaatt
    12061 gatccggctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
    12121 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
    12181 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
    12241 gagtgtatgc tgcacatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
    12301 cttcaagaat taattcccaa ttccccaggc atcaaataaa acgaaaggct cagtcgaaag
    12361 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
    12421 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
    12481 cgccataaac tgccaggaat taattcccca ggcatcaaat aaaacgaaag gctcagtcga
    12541 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
    12601 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
    12661 gcccgccata aactgccagg aattaattcc ccaggcatca aataaaacga aaggctcagt
    12721 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
    12781 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
    12841 gacgcccgcc ataaactgcc aggaattggg gatcg
//
