LOCUS       sequence-309006-       10816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PphlF_A2NT in dCRv1, 2x cascade + [dCas9 + PA4-mVenus].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10816)
  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 10816)
  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: pJK430
FEATURES             Location/Qualifiers
     source          1..10816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /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     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        649..704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    685..703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             722..733
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(740..766,767..769,770..3256,3257..3259,3260..4846)
                     /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      4870..4941
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4957..4984
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     4997..5015
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5019..5062)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_RNA        5145..5220
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5507..5541
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     terminator      6285..6356
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6372..6399
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        6496..6571
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(7179..8129)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      8177..8399
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     misc_RNA        8944..8993
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             9023..9034
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             9043..9756
                     /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             9043..9756
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(9073..9101)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     9696..9716
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      9817..9903
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(9916..10010)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(10031..10690)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     10605..10624
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(10691..10793)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta
      661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcaaaagatc
      721 taaagaggag aaaggatcta tggataagaa atactcaata ggcttagcta tcggcacaaa
      781 tagcgtcgga tgggcggtga tcactgatga atataaggtt ccgtctaaaa agttcaaggt
      841 tctgggaaat acagaccgcc acagtatcaa aaaaaatctt ataggggctc ttttatttga
      901 cagtggagag acagcggaag cgactcgtct caaacggaca gctcgtagaa ggtatacacg
      961 tcggaagaat cgtatttgtt atctacagga gattttttca aatgagatgg cgaaagtaga
     1021 tgatagtttc tttcatcgac ttgaagagtc ttttttggtg gaagaagaca agaagcatga
     1081 acgtcatcct atttttggaa atatagtaga tgaagttgct tatcatgaga aatatccaac
     1141 tatctatcat ctgcgaaaaa aattggtaga ttctactgat aaagcggatt tgcgcttaat
     1201 ctatttggcc ttagcgcata tgattaagtt tcgtggtcat tttttgattg agggagattt
     1261 aaatcctgat aatagtgatg tggacaaact atttatccag ttggtacaaa cctacaatca
     1321 attatttgaa gaaaacccta ttaacgcaag tggagtagat gctaaagcga ttctttctgc
     1381 acgattgagt aaatcaagac gattagaaaa tctcattgct cagctccccg gtgagaagaa
     1441 aaatggctta tttgggaatc tcattgcttt gtcattgggt ttgaccccta attttaaatc
     1501 aaattttgat ttggcagaag atgctaaatt acagctttca aaagatactt acgatgatga
     1561 tttagataat ttattggcgc aaattggaga tcaatatgct gatttgtttt tggcagctaa
     1621 gaatttatca gatgctattt tactttcaga tatcctaaga gtaaatactg aaataactaa
     1681 ggctccccta tcagcttcaa tgattaaacg ctacgatgaa catcatcaag acttgactct
     1741 tttaaaagct ttagttcgac aacaacttcc agaaaagtat aaagaaatct tttttgatca
     1801 atcaaaaaac ggatatgcag gttatattga tgggggagct agccaagaag aattttataa
     1861 atttatcaaa ccaattttag aaaaaatgga tggtactgag gaattattgg tgaaactaaa
     1921 tcgtgaagat ttgctgcgca agcaacggac ctttgacaac ggctctattc cccatcaaat
     1981 tcacttgggt gagctgcatg ctattttgag aagacaagaa gacttttatc catttttaaa
     2041 agacaatcgt gagaagattg aaaaaatctt gacttttcga attccttatt atgttggtcc
     2101 attggcgcgt ggcaatagtc gttttgcatg gatgactcgg aagtctgaag aaacaattac
     2161 cccatggaat tttgaagaag ttgtcgataa aggtgcttca gctcaatcat ttattgaacg
     2221 catgacaaac tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac atagtttgct
     2281 ttatgagtat tttacggttt ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat
     2341 gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc attgttgatt tactcttcaa
     2401 aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg
     2461 ttttgatagt gttgaaattt caggagttga agatagattt aatgcttcat taggtaccta
     2521 ccatgatttg ctaaaaatta ttaaagataa agattttttg gataatgaag aaaatgaaga
     2581 tatcttagag gatattgttt taacattgac cttatttgaa gatagggaga tgattgagga
     2641 aagacttaaa acatatgctc acctctttga tgataaggtg atgaaacagc ttaaacgtcg
     2701 ccgttatact ggttggggac gtttgtctcg aaaattgatt aatggtatta gggataagca
     2761 atctggcaaa acaatattag attttttgaa atcagatggt tttgccaatc gcaattttat
     2821 gcagctgatc catgatgata gtttgacatt taaagaagac attcaaaaag cacaagtgtc
     2881 tggacaaggc gatagtttac atgaacatat tgcaaattta gctggtagcc ctgctattaa
     2941 aaaaggtatt ttacagactg taaaagttgt tgatgaattg gtcaaagtaa tggggcggca
     3001 taagccagaa aatatcgtta ttgaaatggc acgtgaaaat cagacaactc aaaagggcca
     3061 gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat taggaagtca
     3121 gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta
     3181 ttatctccaa aatggaagag acatgtatgt ggaccaagaa ttagatatta atcgtttaag
     3241 tgattatgat gtcgatgcca ttgttccaca aagtttcctt aaagacgatt caatagacaa
     3301 taaggtctta acgcgttctg ataaaaatcg tggtaaatcg gataacgttc caagtgaaga
     3361 agtagtcaaa aagatgaaaa actattggag acaacttcta aacgccaagt taatcactca
     3421 acgtaagttt gataatttaa cgaaagctga acgtggaggt ttgagtgaac ttgataaagc
     3481 tggttttatc aaacgccaat tggttgaaac tcgccaaatc actaagcatg tggcacaaat
     3541 tttggatagt cgcatgaata ctaaatacga tgaaaatgat aaacttattc gagaggttaa
     3601 agtgattacc ttaaaatcta aattagtttc tgacttccga aaagatttcc aattctataa
     3661 agtacgtgag attaacaatt accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg
     3721 aactgctttg attaagaaat atccaaaact tgaatcggag tttgtctatg gtgattataa
     3781 agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca aagcaaccgc
     3841 aaaatatttc ttttactcta atatcatgaa cttcttcaaa acagaaatta cacttgcaaa
     3901 tggagagatt cgcaaacgcc ctctaatcga aactaatggg gaaactggag aaattgtctg
     3961 ggataaaggg cgagattttg ccacagtgcg caaagtattg tccatgcccc aagtcaatat
     4021 tgtcaagaaa acagaagtac agacaggcgg attctccaag gagtcaattt taccaaaaag
     4081 aaattcggac aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt
     4141 tgatagtcca acggtagctt attcagtcct agtggttgct aaggtggaaa aagggaaatc
     4201 gaagaagtta aaatccgtta aagagttact agggatcaca attatggaaa gaagttcctt
     4261 tgaaaaaaat ccgattgact ttttagaagc taaaggatat aaggaagtta aaaaagactt
     4321 aatcattaaa ctacctaaat atagtctttt tgagttagaa aacggtcgta aacggatgct
     4381 ggctagtgcc ggagaattac aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa
     4441 ttttttatat ttagctagtc attatgaaaa gttgaagggt agtccagaag ataacgaaca
     4501 aaaacaattg tttgtggagc agcataagca ttatttagat gagattattg agcaaatcag
     4561 tgaattttct aagcgtgtta ttttagcaga tgccaattta gataaagttc ttagtgcata
     4621 taacaaacat agagacaaac caatacgtga acaagcagaa aatattattc atttatttac
     4681 gttgacgaat cttggagctc ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa
     4741 acgatatacg tctacaaaag aagttttaga tgccactctt atccatcaat ccatcactgg
     4801 tctttatgaa acacgcattg atttgagtca gctaggaggt gactaactcg agtaaggatc
     4861 tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     4921 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     4981 tgcgtttata cctagtaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt
     5041 agctttcgct aaggatgatt tctggaattc taaagatctc agaaatcatt ttacaccaac
     5101 gggtcacacg ggtattatgc tagcataata ccagctagtt gtgggtttta gagctagaaa
     5161 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     5221 tttttttctc ggtaccaaat tccagaaaag aggcctcccg aaaggggggc cttttttcgt
     5281 tttggtccgc ttaagctttt tacacctcca caactagctg gtattatgct agcataatac
     5341 ctaggactga gctgttttag agctagaaat agcaagttaa aataaggcta gtccgattgc
     5401 aacttgaaaa agtggcaccg agtcggtgct ttttttctcg gtaccaaatt ccagaaaaga
     5461 gacgctttcg agcgtctttt ttcgttttgg tccgatagga tccactttta cagctagctc
     5521 agtcctaggt attatgctag cctgaagtac gtctgagcgt gatacccgct cactgaagat
     5581 ggcccggtag ggccgaaacg tacctctaca aataattttg tttaactatg gactatgttt
     5641 gaaagggaga aatactagat ggcacgtacc ccgagccgta gcagcattgg tagcctgcgt
     5701 agtccgcata cccataaagc aattctgacc agcaccattg aaatcctgaa agaatgtggt
     5761 tatagcggtc tgagcattga aagcgttgca cgtcgtgccg gtgcaagcaa accgaccatt
     5821 tatcgttggt ggaccaataa agcagcactg attgccgaag tgtatgaaaa tgaaagcgaa
     5881 caggtgcgta aatttccgga tctgggtagc tttaaagccg atctggattt tctgctgcgt
     5941 aatctgtgga aagtttggcg tgaaaccatt tgtggtgaag catttcgttg tgttattgca
     6001 gaagcacagc tggaccctgc aaccctgacc cagctgaaag atcagtttat ggaacgtcgt
     6061 cgtgagatgc cgaaaaaact ggttgaaaat gccattagca atggtgaact gccgaaagat
     6121 accaatcgtg aactgctgct ggatatgatt tttggttttt gttggtatcg cctgctgacc
     6181 gaacagctga ccgttgaaca ggatattgaa gaatttacct tcctgctgat taatggtgtt
     6241 tgtccgggta cacagcgtta actagggccc atacccccag gcatcaaata aaacgaaagg
     6301 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     6361 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatatacgc gacgtacggt
     6421 ggaatctgat tcgttaccaa ttgacatgat acgaaacgta ccgtatcgtt aaggtataat
     6481 acccgtgtga cccgtgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     6541 tcaacttgaa aaagtggcac cgagtcggtg ctttttttct cggtaccaaa gacgaacaat
     6601 aagacgctga aaagcgtctt ttttcgtttt ggtcctttag aattcctgca ggcttctagg
     6661 gtacgggttt tgctgcccgc aaacgggctg ttctggtgtt gctagtttgt tatcagaatc
     6721 gcagatccgg cttcagccgg tttgccggct gaaagcgcta tttcttccag aattgccatg
     6781 attttttccc cacgggaggc gtcactggct cccgtgttgt cggcagcttt gattcgataa
     6841 gcagcatcgc ctgtttcagg ctgtctatgt gtgactgttg agctgtaaca agttgtctca
     6901 ggtgttcaat ttcatgttct agttgctttg ttttactggt ttcacctgtt ctattaggtg
     6961 ttacatgctg ttcatctgtt acattgtcga tctgttcatg gtgaacagct ttgaatgcac
     7021 caaaaactcg taaaagctct gatgtatcta tcttttttac accgttttca tctgtgcata
     7081 tggacagttt tccctttgat atgtaacggt gaacagttgt tctacttttg tttgttagtc
     7141 ttgatgcttc actgatagat acaagagcca taagaacctc agatccttcc gtatttagcc
     7201 agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga
     7261 tcatacttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt
     7321 gcagttaaag catcgtgtag tgtttttctt agtccgttat gtaggtagga atctgatgta
     7381 atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg
     7441 agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc
     7501 ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa
     7561 acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta
     7621 aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt
     7681 aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga
     7741 ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact
     7801 aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct
     7861 ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct
     7921 aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg
     7981 aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca
     8041 cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca
     8101 tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca
     8161 ctataccaat tgagatgggc tagtcaatga taattactag tccttttcct ttgagttgtg
     8221 ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct
     8281 gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat
     8341 agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca
     8401 ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta
     8461 caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg
     8521 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
     8581 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct
     8641 tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc
     8701 agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt
     8761 cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac
     8821 tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc ttactgtccc
     8881 tagtgcttgg cctagtatgc gtctccttta cacctccaca actagctggt attatgctag
     8941 cagctgtcac cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac
     9001 aaataatttt gtttaacttt acaaagagga gaaaggtacc gcatgagtaa aggagaagaa
     9061 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
     9121 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattg
     9181 atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac tttgggttat
     9241 ggtgttcaat gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt
     9301 gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac
     9361 aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa
     9421 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac
     9481 tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc gaacttcaaa
     9541 attagacaca acattgaaga tggaggtgtt caactagcag accattatca acaaaatact
     9601 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccta ccaatctaag
     9661 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
     9721 gctgggatta cacatggcat ggatgaacta tacaaataag cttgatatcg aattcctgca
     9781 gcccggggga tcccattgta cgcgtgctag aggcatcaaa taaaacgaaa ggctcagtcg
     9841 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     9901 aattctagtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
     9961 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
    10021 cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
    10081 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
    10141 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
    10201 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
    10261 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
    10321 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
    10381 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
    10441 tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg
    10501 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
    10561 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
    10621 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
    10681 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
    10741 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc
    10801 attttcgcca gatatc
//
