LOCUS       sequence-376082-       11648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter circuit for hijacking mRNA of cusCFAB as CRISPR RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11648)
  AUTHORS   Liu Y, Pinto F, Wan X, Yang Z, Peng S, Li M, Cooper JM, Xie Z, 
            French CE, Wang B
  TITLE     Reprogrammed tracrRNAs enable repurposing of RNAs as crRNAs and 
            sequence-specific RNA biosensors.
  JOURNAL   Nat Commun. 2022 Apr 11;13(1):1937. doi: 10.1038/s41467-022-29604-x.
  PUBMED    35410423
REFERENCE   2  (bases 1 to 11648)
  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: "Nat 
            Commun."; date: "2022-04-11"; volume: "13(1):1937. doi"; pages: " 
            10.1038/s41467-022-29604-x"
COMMENT     Sequence Label: pLY320
FEATURES             Location/Qualifiers
     source          1..11648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..12)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(27..653)
                     /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     558..577
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        669..724
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    705..723
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             742..753
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(760..786,787..789,790..3276,3277..3279,3280..4866)
                     /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"
     misc_feature    4930..4951
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      4954..4997
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(5001..5019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5084..5188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5189..5257,5258..6049)
                     /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(5407..5426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6079..6173
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6665..6887
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             6935..7885
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     primer_bind     8147..8164
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(8188..8245)
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     misc_feature    8246..8266
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(8373..9086)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             9093..9104
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(9137..9186)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             complement(9389..9400)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(10398..10516)
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     terminator      complement(10574..10601)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        10617..10651
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     CDS             10669..11505
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
ORIGIN
        1 gcggccttca ataattggct ctagtattaa gacccacttt cacatttaag ttgtttttct
       61 aatccgcata tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca
      121 aataattcga tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt
      181 tcttctttag cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc
      241 acagcgctga gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct
      301 aattgatttt cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt
      361 gctccatcgc gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa
      421 tcttgccagc tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca
      481 atggctaagg cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc
      541 tctacaccta gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta
      601 agcagctcta atgcgctgtt aatcacttta cttttatcta atcttgacat cattaattcc
      661 taatttttgt tgacactcta tcgttgatag agttatttta ccactcccta tcagtgatag
      721 agaaaagaat tgaaaagatc taaagaggag aaaggatcta tggataagaa atactcaata
      781 ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt
      841 ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt
      901 ataggggctc ttttatttga cagtggagag acagcggaag cgactcgcct caaacggaca
      961 gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca
     1021 aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg
     1081 gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct
     1141 tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat
     1201 aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat
     1261 tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag
     1321 ttggtacaaa cctacaatca attatttgaa gaaaacccta ttaacgcaag tggagtagat
     1381 gctaaagcga ttctttctgc acgattgagt aaatcaagac gattagaaaa tctcattgct
     1441 cagctccccg gtgagaagaa aaatggctta tttgggaatc tcattgcttt gtcattgggt
     1501 ttgaccccta attttaaatc aaattttgat ttggcagaag atgctaaatt acagctttca
     1561 aaagatactt acgatgatga tttagataat ttattggcgc aaattggaga tcaatatgct
     1621 gatttgtttt tggcagctaa gaatttatca gatgctattt tactttcaga tatcctaaga
     1681 gtaaatactg aaataactaa ggctccccta tcagcttcaa tgattaaacg ctacgatgaa
     1741 catcatcaag acttgactct tttaaaagct ttagttcgac aacaacttcc agaaaagtat
     1801 aaagaaatct tttttgatca atcaaaaaac ggatatgcag gttatattga tgggggagct
     1861 agccaagaag aattttataa atttatcaaa ccaattttag aaaaaatgga tggtactgag
     1921 gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac ctttgacaac
     1981 ggctctattc cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa
     2041 gacttttatc catttttaaa agacaatcgt gagaagattg aaaaaatctt gacttttcgt
     2101 attccttatt atgttggtcc attggcgcgt ggcaatagtc gttttgcatg gatgactcgg
     2161 aagtctgaag aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca
     2221 gctcaatcat ttattgaacg catgacaaac tttgataaaa atcttccaaa tgaaaaagta
     2281 ctaccaaaac atagtttgct ttatgagtat tttacggttt ataacgaatt gacaaaggtc
     2341 aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc
     2401 attgttgatt tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat
     2461 tatttcaaaa aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt
     2521 aatgcttcat taggtaccta ccatgatttg ctaaaaatta ttaaagataa agattttttg
     2581 gataatgaag aaaatgaaga tatcttagag gatattgttt taacattgac cttatttgaa
     2641 gatagggaga tgattgagga aagacttaaa acatatgctc acctctttga tgataaggtg
     2701 atgaaacagc ttaaacgtcg ccgttatact ggttggggac gtttgtctcg aaaattgatt
     2761 aatggtatta gggataagca atctggcaaa acaatattag attttttgaa atcagatggt
     2821 tttgccaatc gcaattttat gcagctgatc catgatgata gtttgacatt taaagaagac
     2881 attcaaaaag cacaagtgtc tggacaaggc gatagtttac atgaacatat tgcaaattta
     2941 gctggtagcc ctgctattaa aaaaggtatt ttacagactg taaaagttgt tgatgaattg
     3001 gtcaaagtaa tggggcggca taagccagaa aatatcgtta ttgaaatggc acgtgaaaat
     3061 cagacaactc aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt
     3121 atcaaagaat taggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa
     3181 aatgaaaagc tctatctcta ttatctccaa aatggaagag acatgtatgt ggaccaagaa
     3241 ttagatatta atcgtttaag tgattatgat gtcgatgcca ttgttccaca aagtttcctt
     3301 aaagacgatt caatagacaa taaggtctta acgcgttctg ataaaaatcg tggtaaatcg
     3361 gataacgttc caagtgaaga agtagtcaaa aagatgaaaa actattggag acaacttcta
     3421 aacgccaagt taatcactca acgtaagttt gataatttaa cgaaagctga acgtggaggt
     3481 ttgagtgaac ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgccaaatc
     3541 actaagcatg tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat
     3601 aaacttattc gagaggttaa agtgattacc ttaaaatcta aattagtttc tgacttccga
     3661 aaagatttcc aattctataa agtacgtgag attaacaatt accatcatgc ccatgatgcg
     3721 tatctaaatg ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaatcggag
     3781 tttgtctatg gtgattataa agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa
     3841 gaaataggca aagcaaccgc aaaatatttc ttttactcta atatcatgaa cttcttcaaa
     3901 acagaaatta cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga aactaatggg
     3961 gaaactggag aaattgtctg ggataaaggg cgagattttg ccacagtgcg caaagtattg
     4021 tccatgcccc aagtcaatat tgtcaagaaa acagaagtac agacaggcgg attctccaag
     4081 gagtcaattt taccaaaaag aaattcggac aagcttattg ctcgtaaaaa agactgggat
     4141 ccaaaaaaat atggtggttt tgatagtcca acggtagctt attcagtcct agtggttgct
     4201 aaggtggaaa aagggaaatc gaagaagtta aaatccgtta aagagttact agggatcaca
     4261 attatggaaa gaagttcctt tgaaaaaaat ccgattgact ttttagaagc taaaggatat
     4321 aaggaagtta aaaaagactt aatcattaaa ctacctaaat atagtctttt tgagttagaa
     4381 aacggtcgta aacggatgct ggctagtgcc ggagaattac aaaaaggaaa tgagctggct
     4441 ctgccaagca aatatgtgaa ttttttatat ttagctagtc attatgaaaa gttgaagggt
     4501 agtccagaag ataacgaaca aaaacaattg tttgtggagc agcataagca ttatttagat
     4561 gagattattg agcaaatcag tgaattttct aagcgtgtta ttttagcaga tgccaattta
     4621 gataaagttc ttagtgcata taacaaacat agagacaaac caatacgtga acaagcagaa
     4681 aatattattc atttatttac gttgacgaat cttggagctc ccgctgcttt taaatatttt
     4741 gatacaacaa ttgatcgtaa acgatatacg tctacaaaag aagttttaga tgccactctt
     4801 atccatcaat ccatcactgg tctttatgaa acacgcattg atttgagtca gctaggaggt
     4861 gactaatact agagccaatt attgaagggg agcgggaaac cgctcccctt tttttgtttc
     4921 tggtctcccc tctagaagcg gccgcgaatt ccagaaatca tccttagcga aagctaagga
     4981 ttttttttat ctgaaattct gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5041 aataatggtt tcttagacgt caggtggcac catgggaaat gtgcgcggaa cccctatttg
     5101 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5161 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5221 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5281 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5341 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5401 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5461 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5521 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5581 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5641 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5701 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5761 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5821 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5881 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5941 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6001 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6061 agtttacgag ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     6121 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca
     6181 ctagggacag taagacgggt aagcctgttg atgataccgc tgccttactg ggtgcattag
     6241 ccagtctgaa tgacctgtca cgggataatc cgaagtggtc agactggaaa atcagagggc
     6301 aggaactgct gaacagcaaa aagtcagata gcaccacata gcagacccgc cataaaacgc
     6361 cctgagaagc ccgtgacggg cttttcttgt attatgggta gtttccttgc atgaatccat
     6421 aaaaggcgcc tgtagtgcca tttaccccca ttcactgcca gagccgtgag cgcagcgaac
     6481 tgaatgtcac gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa aaggaatatt
     6541 cagcgatttg cccgagcttg cgagggtgct acttaagcct ttagggtttt aaggtctgtt
     6601 ttgtagagga gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa ctgactaaag
     6661 tagtgagtta tacacagggc tgggatctat tctttttatc tttttttatt ctttctttat
     6721 tctataaatt ataaccactt gaatataaac aaaaaaaaca cacaaaggtc tagcggaatt
     6781 tacagagggt ctagcagaat ttacaagttt tccagcaaag gtctagcaga atttacagat
     6841 acccacaact caaaggaaaa ggacatgtaa ttatcattga ctagcccatc tcaattggta
     6901 tagtgattaa aatcacctag accaattgag atgtatgtct gaattagttg ttttcaaagc
     6961 aaatgaacta gcgattagtc gctatgactt aacggagcat gaaaccaagc taattttatg
     7021 ctgtgtggca ctactcaacc ccacgattga aaaccctaca aggaaagaac ggacggtatc
     7081 gttcacttat aaccaatacg ctcagatgat gaacatcagt agggaaaatg cttatggtgt
     7141 attagctaaa gcaaccagag agctgatgac gagaactgtg gaaatcagga atcctttggt
     7201 taaaggcttt gagattttcc agtggacaaa ctatgccaag ttctcaagcg aaaaattaga
     7261 attagttttt agtgaagaga tattgcctta tcttttccag ttaaaaaaat tcataaaata
     7321 taatctggaa catgttaagt cttttgaaaa caaatactct atgaggattt atgagtggtt
     7381 attaaaagaa ctaacacaaa agaaaactca caaggcaaat atagagatta gccttgatga
     7441 atttaagttc atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc ttaaccaatg
     7501 ggttttgaaa ccaataagta aagatttaaa cacttacagc aatatgaaat tggtggttga
     7561 taagcgaggc cgcccgactg atacgttgat tttccaagtt gaactagata gacaaatgga
     7621 tctcgtaacc gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa taccaacaac
     7681 cattacatca gattcctacc tacataacgg actaagaaaa acactacacg atgctttaac
     7741 tgcaaaaatt cagctcacca gttttgaggc aaaatttttg agtgacatgc aaagtaagta
     7801 tgatctcaat ggttcgttct catggctcac gcaaaaacaa cgaaccacac tagagaacat
     7861 actggctaaa tacggaagga tctgaggttc ttatggctct tgtatctatc agtgaagcat
     7921 caagactaac aaacaaaagt agaacaactg ttcaccgtta catatcaaag ggaaaactgt
     7981 ccatatgcac agatgaaaac ggtgtaaaaa agatagatac atcagagctt ttacgagttt
     8041 ttggtgcatt caaagctgtt caccatgaac agatcgacaa tgtaacctcg agattaccgc
     8101 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     8161 cgaggaagcc tgcataacgc gaagtaatct tttcggtttt aaagaaaaag ggcagggtgg
     8221 tgacaccttg cccttttttg ccggactgca gcggccgcta ctagtaggga gaccagaaac
     8281 aaaaaaaggc cccccgttag ggaggccttc aataattggc tctagtatta ttaaactgat
     8341 gcagcgtagt tttcgtcgtt tgcagcaggc cttttgtaca gttcatccat accatgcgtg
     8401 atgcccgctg cggttacgaa ctccagcaga accatatgat cgcgtttctc gttcggatct
     8461 ttagacagaa cgctttgcgt gctcagatag tgattgtctg gcagcagaac aggaccatca
     8521 ccgattggag tgttttgctg gtagtgatca gccagctgca cgctgccatc ctccacgttg
     8581 tggcgaattt taaaattcgc tttaatgcca tttttttgtt tatcggcggt gatgtaaaca
     8641 ttgtggctgt taaaattgta ttccagctta tggcccagga tattgccgtc ttctttaaag
     8701 tcaatgcctt tcagctcaat gcggtttacc agggtatcgc cttcaaattt cacttccgca
     8761 cgcgttttgt acgtgccgtc atccttaaag gaaatcgtgc gttcctgcac atagccttcc
     8821 ggcatggcgg acttgaagaa gtcatgctgc ttcatatggt ccggataacg agcaaagcac
     8881 tgaacaccat aagtcagcgt cgttaccaga gtcggccaag gtaccggcag tttaccagta
     8941 gtacagatga acttcagcgt cagtttacca ttagttgcgt caccttcacc ctcgccacgc
     9001 acggaaaact tatgaccgtt gacatcacca tccagttcca ccagaatagg gacgacacca
     9061 gtgaacagct cttcgccttt acgcatatgg tatttctcct ctttaatctc tagttaaaca
     9121 aaattatttg tagaggctgt ttcgtcctca cggactcatc agaccggaaa gcacatccgg
     9181 tgacagctct ctagtatttg cgttcaggat tgtcctgctg aactgttaat acaatttgcg
     9241 tgccaatttt ttatcttttt gatttataaa gatctgattg aagaatcaac agcaacatgc
     9301 caggatgagt tagcgaatta cactaaccga ctgctacctg tcaacagttc tctagtaggg
     9361 agaccagaaa caaaaaaagg ccgcgttagc ggccttcaat aattggctct agtattaatt
     9421 tgctgctttc cattgagcct gtttctctgc gcgacgttcg cggcggcgtg tgcgtgcatt
     9481 catttgcaaa ctgagttgca gcaactcttt ttcctgctgc atctgaaact cacgtaaatc
     9541 cagcggcagt gttggaagcg aggtggtttc tgaaacggcg atagcgtctt caggcggacg
     9601 ccgtttaaag ggatcaataa tgatgtcatc aagcggataa tcgctggtgc cgtggcgata
     9661 cactgaacgt tccaccacgt ttttcaattc acgaatattt cccggccaac gataattcag
     9721 caatgtttct ctggcgcgct ccgtaaaccc cgggaacaga ggcagcttga tttcccgaca
     9781 catctggatg gcaaagtatt ctgccatcaa cattatgtcg ctttcgcgct cgcgcagtgg
     9841 tggcagttgt acaacatcaa aagccagtcg gtcgagcagg tcagcgcgaa aagtgccttc
     9901 attgaccatc gccgggagat cggcattcgt cgcgcatacc aaccgcacat tcacctgcaa
     9961 tggttggctg ccgccaacgc gctccagttc accgtactca atcacgcgca ataatttctc
    10021 ctgcaccatc atcggtgccg tagcgagttc atcaagaaat agcgtaccgc cgtcggcacg
    10081 ttcaaatctc cctggatgac gtttttgcgc accggtaaac gcccccgctt cgtgaccaaa
    10141 cagttcggaa tccagcagat tttcatttaa cgccgcgcag ttaagggaaa taaacggccc
    10201 ttgccaacgg gaggagagat aatgcaggcg gctggcaatc agctctttac cggtgccgcg
    10261 ttcgccgatg atgagcaccg gtttgtccag cggtgcgaga tgcgaaacct gttccagcac
    10321 ttcgagaaag ctgttcgcct caccaagtaa attatctttg tattctgcca tctagtactt
    10381 tcctgtgtga ctctagttac gaccagtcta aaaagcgcct caattcgcga ccttctcgtt
    10441 actgacagga aaatgggcca ttggcaacca gggaaagatg aacgtgatga tgttcacaat
    10501 ttgctgaatt gtggtgatgt gatgctcacc gcattcctgc tccagcggcg taaaggtctc
    10561 tctagtatat aaacgcagaa aggcccaccc gaaggtgagc cagtgtgata ctagagttta
    10621 cagctagctc agtcctaggt attatgctag ctactagagg gagcccaaat gaccgtatta
    10681 catagtgtgg atttttttcc gtccggtaac gcgtccgtgg cgatagaacc ccggctcccg
    10741 caggcggatt ttcctgaaca tcatcatgat tttcatgaaa ttgtgattgt cgaacatggc
    10801 acgggtattc atgtgtttaa tgggcagccc tataccatca ccggtggcac ggtctgtttc
    10861 gtacgcgatc atgatcggca tctgtatgaa cataccgata atctgtgtct gaccaatgtg
    10921 ctgtatcgct cgccggatcg atttcagttt ctcgccgggc tgaatcagtt gctgccacaa
    10981 gagctggatg ggcagtatcc gtctcactgg cgcgttaacc acagcgtatt gcagcaggtg
    11041 cgacagctgg ttgcacagat ggaacagcag gaaggggaaa atgatttacc ctcgaccgcc
    11101 agtcgcgaga tcttgtttat gcaattactg ctcttgctgc gtaaaagcag tttgcaggag
    11161 aacctggaaa acagcgcatc acgtctcaac ttgcttctgg cctggctgga ggaccatttt
    11221 gccgatgagg tgaattggga tgccgtggcg gatcaatttt ctctttcact gcgtacgcta
    11281 catcggcagc ttaagcagca aacgggactg acgcctcagc gatacctgaa ccgcctgcga
    11341 ctgatgaaag cccgacatct gctacgccac agcgaggcca gcgttactga catcgcctat
    11401 cgctgtggat tcagcgacag taaccacttt tcgacgcttt ttcgccgaga gtttaactgg
    11461 tcaccgcgtg atattcgcca gggacgggat ggctttctgc aataatacta gctcggtacc
    11521 aaagacgaac aataagacgc tgaaaagcgt cttttttcgt tttggtccct ctagtattaa
    11581 ttttgtcttc atgcatgaag acaactaggc tctagtaggg agaccagaaa caaaaaaagg
    11641 ccgcgtta
//
