LOCUS       sequence-376139-       11648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter circuit for hijacking RFP mRNA as CRISPR RNA (target site 
            2).
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: pLY163
FEATURES             Location/Qualifiers
     source          1..11648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(35..46)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(61..687)
                     /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     592..611
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        703..758
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    739..757
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             776..787
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(794..820,821..823,824..3310,3311..3313,3314..4900)
                     /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    4964..4985
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      4988..5031
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(5035..5053)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5118..5222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5223..5291,5292..6083)
                     /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(5441..5460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6113..6207
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6699..6921
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             6969..7919
                     /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     8181..8198
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(8222..8279)
                     /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    8280..8300
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(8407..9120)
                     /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             9127..9138
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(9171..9220)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             complement(9423..9434)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(10432..10550)
                     /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(10608..10635)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        10651..10685
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     CDS             10703..11539
                     /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 gtagggagac cagaaacaaa aaaaggccgc gttagcggcc ttcaataatt ggctctagta
       61 ttaagaccca ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc
      121 gaataagaag gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa
      181 tggcggcata ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg
      241 atcttccaat acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt
      301 ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg
      361 tttttctgta ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc
      421 acatctaaaa cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg
      481 gcaaaagtga gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg
      541 cttatttttt acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt
      601 acgggttgtt aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac
      661 tttactttta tctaatcttg acatcattaa ttcctaattt ttgttgacac tctatcgttg
      721 atagagttat tttaccactc cctatcagtg atagagaaaa gaattgaaaa gatctaaaga
      781 ggagaaagga tctatggata agaaatactc aataggctta gctatcggca caaatagcgt
      841 cggatgggcg gtgatcactg atgaatataa ggttccgtct aaaaagttca aggttctggg
      901 aaatacagac cgccacagta tcaaaaaaaa tcttataggg gctcttttat ttgacagtgg
      961 agagacagcg gaagcgactc gcctcaaacg gacagctcgt agaaggtata cacgtcggaa
     1021 gaatcgtatt tgttatctac aggagatttt ttcaaatgag atggcgaaag tagatgatag
     1081 tttctttcat cgacttgaag agtctttttt ggtggaagaa gacaagaagc atgaacgtca
     1141 tcctattttt ggaaatatag tagatgaagt tgcttatcat gagaaatatc caactatcta
     1201 tcatctgcga aaaaaattgg tagattctac tgataaagcg gatttgcgct taatctattt
     1261 ggccttagcg catatgatta agtttcgtgg tcattttttg attgagggag atttaaatcc
     1321 tgataatagt gatgtggaca aactatttat ccagttggta caaacctaca atcaattatt
     1381 tgaagaaaac cctattaacg caagtggagt agatgctaaa gcgattcttt ctgcacgatt
     1441 gagtaaatca agacgattag aaaatctcat tgctcagctc cccggtgaga agaaaaatgg
     1501 cttatttggg aatctcattg ctttgtcatt gggtttgacc cctaatttta aatcaaattt
     1561 tgatttggca gaagatgcta aattacagct ttcaaaagat acttacgatg atgatttaga
     1621 taatttattg gcgcaaattg gagatcaata tgctgatttg tttttggcag ctaagaattt
     1681 atcagatgct attttacttt cagatatcct aagagtaaat actgaaataa ctaaggctcc
     1741 cctatcagct tcaatgatta aacgctacga tgaacatcat caagacttga ctcttttaaa
     1801 agctttagtt cgacaacaac ttccagaaaa gtataaagaa atcttttttg atcaatcaaa
     1861 aaacggatat gcaggttata ttgatggggg agctagccaa gaagaatttt ataaatttat
     1921 caaaccaatt ttagaaaaaa tggatggtac tgaggaatta ttggtgaaac taaatcgtga
     1981 agatttgctg cgcaagcaac ggacctttga caacggctct attccccatc aaattcactt
     2041 gggtgagctg catgctattt tgagaagaca agaagacttt tatccatttt taaaagacaa
     2101 tcgtgagaag attgaaaaaa tcttgacttt tcgtattcct tattatgttg gtccattggc
     2161 gcgtggcaat agtcgttttg catggatgac tcggaagtct gaagaaacaa ttaccccatg
     2221 gaattttgaa gaagttgtcg ataaaggtgc ttcagctcaa tcatttattg aacgcatgac
     2281 aaactttgat aaaaatcttc caaatgaaaa agtactacca aaacatagtt tgctttatga
     2341 gtattttacg gtttataacg aattgacaaa ggtcaaatat gttactgaag gaatgcgaaa
     2401 accagcattt ctttcaggtg aacagaagaa agccattgtt gatttactct tcaaaacaaa
     2461 tcgaaaagta accgttaagc aattaaaaga agattatttc aaaaaaatag aatgttttga
     2521 tagtgttgaa atttcaggag ttgaagatag atttaatgct tcattaggta cctaccatga
     2581 tttgctaaaa attattaaag ataaagattt tttggataat gaagaaaatg aagatatctt
     2641 agaggatatt gttttaacat tgaccttatt tgaagatagg gagatgattg aggaaagact
     2701 taaaacatat gctcacctct ttgatgataa ggtgatgaaa cagcttaaac gtcgccgtta
     2761 tactggttgg ggacgtttgt ctcgaaaatt gattaatggt attagggata agcaatctgg
     2821 caaaacaata ttagattttt tgaaatcaga tggttttgcc aatcgcaatt ttatgcagct
     2881 gatccatgat gatagtttga catttaaaga agacattcaa aaagcacaag tgtctggaca
     2941 aggcgatagt ttacatgaac atattgcaaa tttagctggt agccctgcta ttaaaaaagg
     3001 tattttacag actgtaaaag ttgttgatga attggtcaaa gtaatggggc ggcataagcc
     3061 agaaaatatc gttattgaaa tggcacgtga aaatcagaca actcaaaagg gccagaaaaa
     3121 ttcgcgagag cgtatgaaac gaatcgaaga aggtatcaaa gaattaggaa gtcagattct
     3181 taaagagcat cctgttgaaa atactcaatt gcaaaatgaa aagctctatc tctattatct
     3241 ccaaaatgga agagacatgt atgtggacca agaattagat attaatcgtt taagtgatta
     3301 tgatgtcgat gccattgttc cacaaagttt ccttaaagac gattcaatag acaataaggt
     3361 cttaacgcgt tctgataaaa atcgtggtaa atcggataac gttccaagtg aagaagtagt
     3421 caaaaagatg aaaaactatt ggagacaact tctaaacgcc aagttaatca ctcaacgtaa
     3481 gtttgataat ttaacgaaag ctgaacgtgg aggtttgagt gaacttgata aagctggttt
     3541 tatcaaacgc caattggttg aaactcgcca aatcactaag catgtggcac aaattttgga
     3601 tagtcgcatg aatactaaat acgatgaaaa tgataaactt attcgagagg ttaaagtgat
     3661 taccttaaaa tctaaattag tttctgactt ccgaaaagat ttccaattct ataaagtacg
     3721 tgagattaac aattaccatc atgcccatga tgcgtatcta aatgccgtcg ttggaactgc
     3781 tttgattaag aaatatccaa aacttgaatc ggagtttgtc tatggtgatt ataaagttta
     3841 tgatgttcgt aaaatgattg ctaagtctga gcaagaaata ggcaaagcaa ccgcaaaata
     3901 tttcttttac tctaatatca tgaacttctt caaaacagaa attacacttg caaatggaga
     3961 gattcgcaaa cgccctctaa tcgaaactaa tggggaaact ggagaaattg tctgggataa
     4021 agggcgagat tttgccacag tgcgcaaagt attgtccatg ccccaagtca atattgtcaa
     4081 gaaaacagaa gtacagacag gcggattctc caaggagtca attttaccaa aaagaaattc
     4141 ggacaagctt attgctcgta aaaaagactg ggatccaaaa aaatatggtg gttttgatag
     4201 tccaacggta gcttattcag tcctagtggt tgctaaggtg gaaaaaggga aatcgaagaa
     4261 gttaaaatcc gttaaagagt tactagggat cacaattatg gaaagaagtt cctttgaaaa
     4321 aaatccgatt gactttttag aagctaaagg atataaggaa gttaaaaaag acttaatcat
     4381 taaactacct aaatatagtc tttttgagtt agaaaacggt cgtaaacgga tgctggctag
     4441 tgccggagaa ttacaaaaag gaaatgagct ggctctgcca agcaaatatg tgaatttttt
     4501 atatttagct agtcattatg aaaagttgaa gggtagtcca gaagataacg aacaaaaaca
     4561 attgtttgtg gagcagcata agcattattt agatgagatt attgagcaaa tcagtgaatt
     4621 ttctaagcgt gttattttag cagatgccaa tttagataaa gttcttagtg catataacaa
     4681 acatagagac aaaccaatac gtgaacaagc agaaaatatt attcatttat ttacgttgac
     4741 gaatcttgga gctcccgctg cttttaaata ttttgataca acaattgatc gtaaacgata
     4801 tacgtctaca aaagaagttt tagatgccac tcttatccat caatccatca ctggtcttta
     4861 tgaaacacgc attgatttga gtcagctagg aggtgactaa tactagagcc aattattgaa
     4921 ggggagcggg aaaccgctcc cctttttttg tttctggtct cccctctaga agcggccgcg
     4981 aattccagaa atcatcctta gcgaaagcta aggatttttt ttatctgaaa ttctgcctcg
     5041 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5101 gcaccatggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5161 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5221 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5281 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5341 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5401 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5461 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5521 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5581 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5641 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5701 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5761 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5821 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5881 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5941 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6001 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6061 cctcactgat taagcattgg taactgtcag accaagttta cgagctcgct tggactcctg
     6121 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     6181 cgccgggcgt tttttattgg tgagaatcca agcactaggg acagtaagac gggtaagcct
     6241 gttgatgata ccgctgcctt actgggtgca ttagccagtc tgaatgacct gtcacgggat
     6301 aatccgaagt ggtcagactg gaaaatcaga gggcaggaac tgctgaacag caaaaagtca
     6361 gatagcacca catagcagac ccgccataaa acgccctgag aagcccgtga cgggcttttc
     6421 ttgtattatg ggtagtttcc ttgcatgaat ccataaaagg cgcctgtagt gccatttacc
     6481 cccattcact gccagagccg tgagcgcagc gaactgaatg tcacgaaaaa gacagcgact
     6541 caggtgcctg atggtcggag acaaaaggaa tattcagcga tttgcccgag cttgcgaggg
     6601 tgctacttaa gcctttaggg ttttaaggtc tgttttgtag aggagcaaac agcgtttgcg
     6661 acatcctttt gtaatactgc ggaactgact aaagtagtga gttatacaca gggctgggat
     6721 ctattctttt tatctttttt tattctttct ttattctata aattataacc acttgaatat
     6781 aaacaaaaaa aacacacaaa ggtctagcgg aatttacaga gggtctagca gaatttacaa
     6841 gttttccagc aaaggtctag cagaatttac agatacccac aactcaaagg aaaaggacat
     6901 gtaattatca ttgactagcc catctcaatt ggtatagtga ttaaaatcac ctagaccaat
     6961 tgagatgtat gtctgaatta gttgttttca aagcaaatga actagcgatt agtcgctatg
     7021 acttaacgga gcatgaaacc aagctaattt tatgctgtgt ggcactactc aaccccacga
     7081 ttgaaaaccc tacaaggaaa gaacggacgg tatcgttcac ttataaccaa tacgctcaga
     7141 tgatgaacat cagtagggaa aatgcttatg gtgtattagc taaagcaacc agagagctga
     7201 tgacgagaac tgtggaaatc aggaatcctt tggttaaagg ctttgagatt ttccagtgga
     7261 caaactatgc caagttctca agcgaaaaat tagaattagt ttttagtgaa gagatattgc
     7321 cttatctttt ccagttaaaa aaattcataa aatataatct ggaacatgtt aagtcttttg
     7381 aaaacaaata ctctatgagg atttatgagt ggttattaaa agaactaaca caaaagaaaa
     7441 ctcacaaggc aaatatagag attagccttg atgaatttaa gttcatgtta atgcttgaaa
     7501 ataactacca tgagtttaaa aggcttaacc aatgggtttt gaaaccaata agtaaagatt
     7561 taaacactta cagcaatatg aaattggtgg ttgataagcg aggccgcccg actgatacgt
     7621 tgattttcca agttgaacta gatagacaaa tggatctcgt aaccgaactt gagaacaacc
     7681 agataaaaat gaatggtgac aaaataccaa caaccattac atcagattcc tacctacata
     7741 acggactaag aaaaacacta cacgatgctt taactgcaaa aattcagctc accagttttg
     7801 aggcaaaatt tttgagtgac atgcaaagta agtatgatct caatggttcg ttctcatggc
     7861 tcacgcaaaa acaacgaacc acactagaga acatactggc taaatacgga aggatctgag
     7921 gttcttatgg ctcttgtatc tatcagtgaa gcatcaagac taacaaacaa aagtagaaca
     7981 actgttcacc gttacatatc aaagggaaaa ctgtccatat gcacagatga aaacggtgta
     8041 aaaaagatag atacatcaga gcttttacga gtttttggtg cattcaaagc tgttcaccat
     8101 gaacagatcg acaatgtaac ctcgagatta ccgcctttga gtgagctgat accgctcgcc
     8161 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcctgcata acgcgaagta
     8221 atcttttcgg ttttaaagaa aaagggcagg gtggtgacac cttgcccttt tttgccggac
     8281 tgcagcggcc gctactagta gggagaccag aaacaaaaaa aggccccccg ttagggaggc
     8341 cttcaataat tggctctagt attattaaac tgatgcagcg tagttttcgt cgtttgcagc
     8401 aggccttttg tacagttcat ccataccatg cgtgatgccc gctgcggtta cgaactccag
     8461 cagaaccata tgatcgcgtt tctcgttcgg atctttagac agaacgcttt gcgtgctcag
     8521 atagtgattg tctggcagca gaacaggacc atcaccgatt ggagtgtttt gctggtagtg
     8581 atcagccagc tgcacgctgc catcctccac gttgtggcga attttaaaat tcgctttaat
     8641 gccatttttt tgtttatcgg cggtgatgta aacattgtgg ctgttaaaat tgtattccag
     8701 cttatggccc aggatattgc cgtcttcttt aaagtcaatg cctttcagct caatgcggtt
     8761 taccagggta tcgccttcaa atttcacttc cgcacgcgtt ttgtacgtgc cgtcatcctt
     8821 aaaggaaatc gtgcgttcct gcacatagcc ttccggcatg gcggacttga agaagtcatg
     8881 ctgcttcata tggtccggat aacgagcaaa gcactgaaca ccataagtca gcgtcgttac
     8941 cagagtcggc caaggtaccg gcagtttacc agtagtacag atgaacttca gcgtcagttt
     9001 accattagtt gcgtcacctt caccctcgcc acgcacggaa aacttatgac cgttgacatc
     9061 accatccagt tccaccagaa tagggacgac accagtgaac agctcttcgc ctttacgcat
     9121 atggtatttc tcctctttaa tctctagtta aacaaaatta tttgtagagg ctgtttcgtc
     9181 ctcacggact catcagaccg gaaagcacat ccggtgacag ctctctagta tttgcgttca
     9241 ggattgtcct gctgaactgt taatacaatt tgcgtgccaa ttttttatct ttttgattta
     9301 taaagatctg attgaagaat caacagcaac atgccaggat gagttagcga attacactaa
     9361 ccgcatggtc ttcttctgca ttactctagt agggagacca gaaacaaaaa aaggccgcgt
     9421 tagcggcctt caataattgg ctctagtatt aatttgctgc tttccattga gcctgtttct
     9481 ctgcgcgacg ttcgcggcgg cgtgtgcgtg cattcatttg caaactgagt tgcagcaact
     9541 ctttttcctg ctgcatctga aactcacgta aatccagcgg cagtgttgga agcgaggtgg
     9601 tttctgaaac ggcgatagcg tcttcaggcg gacgccgttt aaagggatca ataatgatgt
     9661 catcaagcgg ataatcgctg gtgccgtggc gatacactga acgttccacc acgtttttca
     9721 attcacgaat atttcccggc caacgataat tcagcaatgt ttctctggcg cgctccgtaa
     9781 accccgggaa cagaggcagc ttgatttccc gacacatctg gatggcaaag tattctgcca
     9841 tcaacattat gtcgctttcg cgctcgcgca gtggtggcag ttgtacaaca tcaaaagcca
     9901 gtcggtcgag caggtcagcg cgaaaagtgc cttcattgac catcgccggg agatcggcat
     9961 tcgtcgcgca taccaaccgc acattcacct gcaatggttg gctgccgcca acgcgctcca
    10021 gttcaccgta ctcaatcacg cgcaataatt tctcctgcac catcatcggt gccgtagcga
    10081 gttcatcaag aaatagcgta ccgccgtcgg cacgttcaaa tctccctgga tgacgttttt
    10141 gcgcaccggt aaacgccccc gcttcgtgac caaacagttc ggaatccagc agattttcat
    10201 ttaacgccgc gcagttaagg gaaataaacg gcccttgcca acgggaggag agataatgca
    10261 ggcggctggc aatcagctct ttaccggtgc cgcgttcgcc gatgatgagc accggtttgt
    10321 ccagcggtgc gagatgcgaa acctgttcca gcacttcgag aaagctgttc gcctcaccaa
    10381 gtaaattatc tttgtattct gccatctagt actttcctgt gtgactctag ttacgaccag
    10441 tctaaaaagc gcctcaattc gcgaccttct cgttactgac aggaaaatgg gccattggca
    10501 accagggaaa gatgaacgtg atgatgttca caatttgctg aattgtggtg atgtgatgct
    10561 caccgcattc ctgctccagc ggcgtaaagg tctctctagt atataaacgc agaaaggccc
    10621 acccgaaggt gagccagtgt gatactagag tttacagcta gctcagtcct aggtattatg
    10681 ctagctacta gagggagccc aaatgaccgt attacatagt gtggattttt ttccgtccgg
    10741 taacgcgtcc gtggcgatag aaccccggct cccgcaggcg gattttcctg aacatcatca
    10801 tgattttcat gaaattgtga ttgtcgaaca tggcacgggt attcatgtgt ttaatgggca
    10861 gccctatacc atcaccggtg gcacggtctg tttcgtacgc gatcatgatc ggcatctgta
    10921 tgaacatacc gataatctgt gtctgaccaa tgtgctgtat cgctcgccgg atcgatttca
    10981 gtttctcgcc gggctgaatc agttgctgcc acaagagctg gatgggcagt atccgtctca
    11041 ctggcgcgtt aaccacagcg tattgcagca ggtgcgacag ctggttgcac agatggaaca
    11101 gcaggaaggg gaaaatgatt taccctcgac cgccagtcgc gagatcttgt ttatgcaatt
    11161 actgctcttg ctgcgtaaaa gcagtttgca ggagaacctg gaaaacagcg catcacgtct
    11221 caacttgctt ctggcctggc tggaggacca ttttgccgat gaggtgaatt gggatgccgt
    11281 ggcggatcaa ttttctcttt cactgcgtac gctacatcgg cagcttaagc agcaaacggg
    11341 actgacgcct cagcgatacc tgaaccgcct gcgactgatg aaagcccgac atctgctacg
    11401 ccacagcgag gccagcgtta ctgacatcgc ctatcgctgt ggattcagcg acagtaacca
    11461 cttttcgacg ctttttcgcc gagagtttaa ctggtcaccg cgtgatattc gccagggacg
    11521 ggatggcttt ctgcaataat actagctcgg taccaaagac gaacaataag acgctgaaaa
    11581 gcgtcttttt tcgttttggt ccctctagta ttaattttgt cttcatgcat gaagacaact
    11641 aggctcta
//
