LOCUS       sequence-258306-       10475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A CRISPR activation device with the necessary genes (dcas9 
            controlled by Ptet, pspF-Delta-HTH::lambda-N22plus controlled by 
            promoter J23106), and the reporter part (PpspA-2G6 with 
            sfgfp::ASV)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10475)
  AUTHORS   Liu Y, Wan X, Wang B
  TITLE     Engineered CRISPRa enables programmable eukaryote-like gene 
            activation in bacteria.
  JOURNAL   Nat Commun. 2019 Aug 26;10(1):3693. doi: 10.1038/s41467-019-11479-0.
  PUBMED    31451697
REFERENCE   2  (bases 1 to 10475)
  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: "2019-08-26"; volume: "10(1):3693. doi"; pages: " 
            10.1038/s41467-019-11479-0"
COMMENT     Sequence Label: pLY9
FEATURES             Location/Qualifiers
     source          1..10475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             468..479
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(512..561)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             complement(767..778)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(793..1419)
                     /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     1324..1343
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        1435..1490
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1471..1489
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1508..1519
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1526..1552,1553..1555,1556..4042,4043..4045,
                     4046..5632)
                     /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    5696..5717
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      5720..5763
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(5767..5785)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5850..5954
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5955..6023,6024..6815)
                     /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(6173..6192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6845..6939
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      7431..7653
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             7701..8651
                     /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     8913..8930
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(8954..9011)
                     /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    9012..9032
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(9064..9075)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(10074..10108)
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             complement(join(10223..10475,1..461))
                     /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"
ORIGIN
        1 gcggtgatgt aaacattgtg gctgttaaaa ttgtattcca gcttatggcc caggatattg
       61 ccgtcttctt taaagtcaat gcctttcagc tcaatgcggt ttaccagggt atcgccttca
      121 aatttcactt ccgcacgcgt tttgtacgtg ccgtcatcct taaaggaaat cgtgcgttcc
      181 tgcacatagc cttccggcat ggcggacttg aagaagtcat gctgcttcat atggtccgga
      241 taacgagcaa agcactgaac accataagtc agcgtcgtta ccagagtcgg ccaaggtacc
      301 ggcagtttac cagtagtaca gatgaacttc agcgtcagtt taccattagt tgcgtcacct
      361 tcaccctcgc cacgcacgga aaacttatga ccgttgacat caccatccag ttccaccaga
      421 atagggacga caccagtgaa cagctcttcg cctttacgca tatggtattt ctcctcttta
      481 atctctagtt aaacaaaatt atttgtagag gctgtttcgt cctcacggac tcatcagacc
      541 ggaaagcaca tccggtgaca gctctctagt atttgcgttc aggattgtcc tgctgaactg
      601 ttaatacaat ttgcgtgcca attttttatc tttttgattt ataaagatct gattgaagaa
      661 tcaacagcaa catgccagga tgccgttttt tgccacaaca gctttccgtt ttttgccaca
      721 acagcttttc tagtagggag accagaaaca aaaaaaggcc gcgttagcgg ccttcaataa
      781 ttggctctag tattaagacc cactttcaca tttaagttgt ttttctaatc cgcatatgat
      841 caattcaagg ccgaataaga aggctggctc tgcaccttgg tgatcaaata attcgatagc
      901 ttgtcgtaat aatggcggca tactatcagt agtaggtgtt tccctttctt ctttagcgac
      961 ttgatgctct tgatcttcca atacgcaacc taaagtaaaa tgccccacag cgctgagtgc
     1021 atataatgca ttctctagtg aaaaaccttg ttggcataaa aaggctaatt gattttcgag
     1081 agtttcatac tgtttttctg taggccgtgt acctaaatgt acttttgctc catcgcgatg
     1141 acttagtaaa gcacatctaa aacttttagc gttattacgt aaaaaatctt gccagctttc
     1201 cccttctaaa gggcaaaagt gagtatggtg cctatctaac atctcaatgg ctaaggcgtc
     1261 gagcaaagcc cgcttatttt ttacatgcca atacaatgta ggctgctcta cacctagctt
     1321 ctgggcgagt ttacgggttg ttaaaccttc gattccgacc tcattaagca gctctaatgc
     1381 gctgttaatc actttacttt tatctaatct tgacatcatt aattcctaat ttttgttgac
     1441 actctatcgt tgatagagtt attttaccac tccctatcag tgatagagaa aagaattgaa
     1501 aagatctaaa gaggagaaag gatctatgga taagaaatac tcaataggct tagctatcgg
     1561 cacaaatagc gtcggatggg cggtgatcac tgatgaatat aaggttccgt ctaaaaagtt
     1621 caaggttctg ggaaatacag accgccacag tatcaaaaaa aatcttatag gggctctttt
     1681 atttgacagt ggagagacag cggaagcgac tcgcctcaaa cggacagctc gtagaaggta
     1741 tacacgtcgg aagaatcgta tttgttatct acaggagatt ttttcaaatg agatggcgaa
     1801 agtagatgat agtttctttc atcgacttga agagtctttt ttggtggaag aagacaagaa
     1861 gcatgaacgt catcctattt ttggaaatat agtagatgaa gttgcttatc atgagaaata
     1921 tccaactatc tatcatctgc gaaaaaaatt ggtagattct actgataaag cggatttgcg
     1981 cttaatctat ttggccttag cgcatatgat taagtttcgt ggtcattttt tgattgaggg
     2041 agatttaaat cctgataata gtgatgtgga caaactattt atccagttgg tacaaaccta
     2101 caatcaatta tttgaagaaa accctattaa cgcaagtgga gtagatgcta aagcgattct
     2161 ttctgcacga ttgagtaaat caagacgatt agaaaatctc attgctcagc tccccggtga
     2221 gaagaaaaat ggcttatttg ggaatctcat tgctttgtca ttgggtttga cccctaattt
     2281 taaatcaaat tttgatttgg cagaagatgc taaattacag ctttcaaaag atacttacga
     2341 tgatgattta gataatttat tggcgcaaat tggagatcaa tatgctgatt tgtttttggc
     2401 agctaagaat ttatcagatg ctattttact ttcagatatc ctaagagtaa atactgaaat
     2461 aactaaggct cccctatcag cttcaatgat taaacgctac gatgaacatc atcaagactt
     2521 gactctttta aaagctttag ttcgacaaca acttccagaa aagtataaag aaatcttttt
     2581 tgatcaatca aaaaacggat atgcaggtta tattgatggg ggagctagcc aagaagaatt
     2641 ttataaattt atcaaaccaa ttttagaaaa aatggatggt actgaggaat tattggtgaa
     2701 actaaatcgt gaagatttgc tgcgcaagca acggaccttt gacaacggct ctattcccca
     2761 tcaaattcac ttgggtgagc tgcatgctat tttgagaaga caagaagact tttatccatt
     2821 tttaaaagac aatcgtgaga agattgaaaa aatcttgact tttcgtattc cttattatgt
     2881 tggtccattg gcgcgtggca atagtcgttt tgcatggatg actcggaagt ctgaagaaac
     2941 aattacccca tggaattttg aagaagttgt cgataaaggt gcttcagctc aatcatttat
     3001 tgaacgcatg acaaactttg ataaaaatct tccaaatgaa aaagtactac caaaacatag
     3061 tttgctttat gagtatttta cggtttataa cgaattgaca aaggtcaaat atgttactga
     3121 aggaatgcga aaaccagcat ttctttcagg tgaacagaag aaagccattg ttgatttact
     3181 cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa gaagattatt tcaaaaaaat
     3241 agaatgtttt gatagtgttg aaatttcagg agttgaagat agatttaatg cttcattagg
     3301 tacctaccat gatttgctaa aaattattaa agataaagat tttttggata atgaagaaaa
     3361 tgaagatatc ttagaggata ttgttttaac attgacctta tttgaagata gggagatgat
     3421 tgaggaaaga cttaaaacat atgctcacct ctttgatgat aaggtgatga aacagcttaa
     3481 acgtcgccgt tatactggtt ggggacgttt gtctcgaaaa ttgattaatg gtattaggga
     3541 taagcaatct ggcaaaacaa tattagattt tttgaaatca gatggttttg ccaatcgcaa
     3601 ttttatgcag ctgatccatg atgatagttt gacatttaaa gaagacattc aaaaagcaca
     3661 agtgtctgga caaggcgata gtttacatga acatattgca aatttagctg gtagccctgc
     3721 tattaaaaaa ggtattttac agactgtaaa agttgttgat gaattggtca aagtaatggg
     3781 gcggcataag ccagaaaata tcgttattga aatggcacgt gaaaatcaga caactcaaaa
     3841 gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa gaaggtatca aagaattagg
     3901 aagtcagatt cttaaagagc atcctgttga aaatactcaa ttgcaaaatg aaaagctcta
     3961 tctctattat ctccaaaatg gaagagacat gtatgtggac caagaattag atattaatcg
     4021 tttaagtgat tatgatgtcg atgccattgt tccacaaagt ttccttaaag acgattcaat
     4081 agacaataag gtcttaacgc gttctgataa aaatcgtggt aaatcggata acgttccaag
     4141 tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa cttctaaacg ccaagttaat
     4201 cactcaacgt aagtttgata atttaacgaa agctgaacgt ggaggtttga gtgaacttga
     4261 taaagctggt tttatcaaac gccaattggt tgaaactcgc caaatcacta agcatgtggc
     4321 acaaattttg gatagtcgca tgaatactaa atacgatgaa aatgataaac ttattcgaga
     4381 ggttaaagtg attaccttaa aatctaaatt agtttctgac ttccgaaaag atttccaatt
     4441 ctataaagta cgtgagatta acaattacca tcatgcccat gatgcgtatc taaatgccgt
     4501 cgttggaact gctttgatta agaaatatcc aaaacttgaa tcggagtttg tctatggtga
     4561 ttataaagtt tatgatgttc gtaaaatgat tgctaagtct gagcaagaaa taggcaaagc
     4621 aaccgcaaaa tatttctttt actctaatat catgaacttc ttcaaaacag aaattacact
     4681 tgcaaatgga gagattcgca aacgccctct aatcgaaact aatggggaaa ctggagaaat
     4741 tgtctgggat aaagggcgag attttgccac agtgcgcaaa gtattgtcca tgccccaagt
     4801 caatattgtc aagaaaacag aagtacagac aggcggattc tccaaggagt caattttacc
     4861 aaaaagaaat tcggacaagc ttattgctcg taaaaaagac tgggatccaa aaaaatatgg
     4921 tggttttgat agtccaacgg tagcttattc agtcctagtg gttgctaagg tggaaaaagg
     4981 gaaatcgaag aagttaaaat ccgttaaaga gttactaggg atcacaatta tggaaagaag
     5041 ttcctttgaa aaaaatccga ttgacttttt agaagctaaa ggatataagg aagttaaaaa
     5101 agacttaatc attaaactac ctaaatatag tctttttgag ttagaaaacg gtcgtaaacg
     5161 gatgctggct agtgccggag aattacaaaa aggaaatgag ctggctctgc caagcaaata
     5221 tgtgaatttt ttatatttag ctagtcatta tgaaaagttg aagggtagtc cagaagataa
     5281 cgaacaaaaa caattgtttg tggagcagca taagcattat ttagatgaga ttattgagca
     5341 aatcagtgaa ttttctaagc gtgttatttt agcagatgcc aatttagata aagttcttag
     5401 tgcatataac aaacatagag acaaaccaat acgtgaacaa gcagaaaata ttattcattt
     5461 atttacgttg acgaatcttg gagctcccgc tgcttttaaa tattttgata caacaattga
     5521 tcgtaaacga tatacgtcta caaaagaagt tttagatgcc actcttatcc atcaatccat
     5581 cactggtctt tatgaaacac gcattgattt gagtcagcta ggaggtgact aatactagag
     5641 ccaattattg aaggggagcg ggaaaccgct cccctttttt tgtttctggt ctcccctcta
     5701 gaagcggccg cgaattccag aaatcatcct tagcgaaagc taaggatttt ttttatctga
     5761 aattctgcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5821 agacgtcagg tggcaccatg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5881 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5941 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6001 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6061 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6121 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6181 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6241 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6301 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6361 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6421 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6481 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6541 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6601 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6661 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6721 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6781 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tacgagctcg
     6841 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     6901 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagcactag ggacagtaag
     6961 acgggtaagc ctgttgatga taccgctgcc ttactgggtg cattagccag tctgaatgac
     7021 ctgtcacggg ataatccgaa gtggtcagac tggaaaatca gagggcagga actgctgaac
     7081 agcaaaaagt cagatagcac cacatagcag acccgccata aaacgccctg agaagcccgt
     7141 gacgggcttt tcttgtatta tgggtagttt ccttgcatga atccataaaa ggcgcctgta
     7201 gtgccattta cccccattca ctgccagagc cgtgagcgca gcgaactgaa tgtcacgaaa
     7261 aagacagcga ctcaggtgcc tgatggtcgg agacaaaagg aatattcagc gatttgcccg
     7321 agcttgcgag ggtgctactt aagcctttag ggttttaagg tctgttttgt agaggagcaa
     7381 acagcgtttg cgacatcctt ttgtaatact gcggaactga ctaaagtagt gagttataca
     7441 cagggctggg atctattctt tttatctttt tttattcttt ctttattcta taaattataa
     7501 ccacttgaat ataaacaaaa aaaacacaca aaggtctagc ggaatttaca gagggtctag
     7561 cagaatttac aagttttcca gcaaaggtct agcagaattt acagataccc acaactcaaa
     7621 ggaaaaggac atgtaattat cattgactag cccatctcaa ttggtatagt gattaaaatc
     7681 acctagacca attgagatgt atgtctgaat tagttgtttt caaagcaaat gaactagcga
     7741 ttagtcgcta tgacttaacg gagcatgaaa ccaagctaat tttatgctgt gtggcactac
     7801 tcaaccccac gattgaaaac cctacaagga aagaacggac ggtatcgttc acttataacc
     7861 aatacgctca gatgatgaac atcagtaggg aaaatgctta tggtgtatta gctaaagcaa
     7921 ccagagagct gatgacgaga actgtggaaa tcaggaatcc tttggttaaa ggctttgaga
     7981 ttttccagtg gacaaactat gccaagttct caagcgaaaa attagaatta gtttttagtg
     8041 aagagatatt gccttatctt ttccagttaa aaaaattcat aaaatataat ctggaacatg
     8101 ttaagtcttt tgaaaacaaa tactctatga ggatttatga gtggttatta aaagaactaa
     8161 cacaaaagaa aactcacaag gcaaatatag agattagcct tgatgaattt aagttcatgt
     8221 taatgcttga aaataactac catgagttta aaaggcttaa ccaatgggtt ttgaaaccaa
     8281 taagtaaaga tttaaacact tacagcaata tgaaattggt ggttgataag cgaggccgcc
     8341 cgactgatac gttgattttc caagttgaac tagatagaca aatggatctc gtaaccgaac
     8401 ttgagaacaa ccagataaaa atgaatggtg acaaaatacc aacaaccatt acatcagatt
     8461 cctacctaca taacggacta agaaaaacac tacacgatgc tttaactgca aaaattcagc
     8521 tcaccagttt tgaggcaaaa tttttgagtg acatgcaaag taagtatgat ctcaatggtt
     8581 cgttctcatg gctcacgcaa aaacaacgaa ccacactaga gaacatactg gctaaatacg
     8641 gaaggatctg aggttcttat ggctcttgta tctatcagtg aagcatcaag actaacaaac
     8701 aaaagtagaa caactgttca ccgttacata tcaaagggaa aactgtccat atgcacagat
     8761 gaaaacggtg taaaaaagat agatacatca gagcttttac gagtttttgg tgcattcaaa
     8821 gctgttcacc atgaacagat cgacaatgta acctcgagat taccgccttt gagtgagctg
     8881 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcctgca
     8941 taacgcgaag taatcttttc ggttttaaag aaaaagggca gggtggtgac accttgccct
     9001 tttttgccgg actgcagcgg ccgctactag tagggagacc agaaacaaaa aaaggccgcg
     9061 ttagcggcct tcaataattg gctctagtat taatttgctg ctttccattg agcctgtttc
     9121 tctgcgcgac gttcgcggcg gcgtgtgcgt gcattcattt gcaaactgag ttgcagcaac
     9181 tctttttcct gctgcatctg aaactcacgt aaatccagcg gcagtgttgg aagcgaggtg
     9241 gtttctgaaa cggcgatagc gtcttcaggc ggacgccgtt taaagggatc aataatgatg
     9301 tcatcaagcg gataatcgct ggtgccgtgg cgatacactg aacgttccac cacgtttttc
     9361 aattcacgaa tatttcccgg ccaacgataa ttcagcaatg tttctctggc gcgctccgta
     9421 aaccccggga acagaggcag cttgatttcc cgacacatct ggatggcaaa gtattctgcc
     9481 atcaacatta tgtcgctttc gcgctcgcgc agtggtggca gttgtacaac atcaaaagcc
     9541 agtcggtcga gcaggtcagc gcgaaaagtg ccttcattga ccatcgccgg gagatcggca
     9601 ttcgtcgcgc ataccaaccg cacattcacc tgcaatggtt ggctgccgcc aacgcgctcc
     9661 agttcaccgt actcaatcac gcgcaataat ttctcctgca ccatcatcgg tgccgtagcg
     9721 agttcatcaa gaaatagcgt accgccgtcg gcacgttcaa atctccctgg atgacgtttt
     9781 tgcgcaccgg taaacgcccc cgcttcgtga ccaaacagtt cggaatccag cagattttca
     9841 tttaacgccg cgcagttaag ggaaataaac ggcccttgcc aacgggagga gagataatgc
     9901 aggcggctgg caatcagctc tttaccggtg ccgcgttcgc cgatgatgag caccggtttg
     9961 tccagcggtg cgagatgcga aacctgttcc agcacttcga gaaagctgtt cgcctcacca
    10021 agtaaattat ctttgtattc tgccatctag tactttcctg tgtgactcta gtagctagca
    10081 ctatacctag gactgagcta gccgtaaact ctagtaggga gaccagaaac aaaaaaaggc
    10141 cccccgttag ggaggccttc aataattggc tctagtatta ttaaactgat gcagcgtagt
    10201 tttcgtcgtt tgcagcaggc cttttgtaca gttcatccat accatgcgtg atgcccgctg
    10261 cggttacgaa ctccagcaga accatatgat cgcgtttctc gttcggatct ttagacagaa
    10321 cgctttgcgt gctcagatag tgattgtctg gcagcagaac aggaccatca ccgattggag
    10381 tgttttgctg gtagtgatca gccagctgca cgctgccatc ctccacgttg tggcgaattt
    10441 taaaattcgc tttaatgcca tttttttgtt tatcg
//
