LOCUS       sequence-257328-       10763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A CRISPR activation device with the necessary genes (dcas9 
            controlled by Ptet, pspF-Delta-HTH::MCP 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 10763)
  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 10763)
  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: pLY59
FEATURES             Location/Qualifiers
     source          1..10763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(108..119)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(134..760)
                     /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     665..684
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        776..831
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    812..830
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             849..860
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(867..893,894..896,897..3383,3384..3386,3387..4973)
                     /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    5037..5058
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      5061..5104
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(5108..5126)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5191..5295
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5296..5364,5365..6156)
                     /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(5514..5533)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6186..6280
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6772..6994
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             7042..7992
                     /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     8254..8271
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(8295..8352)
                     /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    8353..8373
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(8405..8416)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(9703..9737)
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             complement(9852..10565)
                     /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             10572..10583
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(10616..10665)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 atcaacagca acatgccagg atgccgtttt ttgccacaac agctttccgt tttttgccac
       61 aacagctttt ctagtaggga gaccagaaac aaaaaaaggc cgcgttagcg gccttcaata
      121 attggctcta gtattaagac ccactttcac atttaagttg tttttctaat ccgcatatga
      181 tcaattcaag gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag
      241 cttgtcgtaa taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga
      301 cttgatgctc ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg
      361 catataatgc attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga
      421 gagtttcata ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat
      481 gacttagtaa agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt
      541 ccccttctaa agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt
      601 cgagcaaagc ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct
      661 tctgggcgag tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg
      721 cgctgttaat cactttactt ttatctaatc ttgacatcat taattcctaa tttttgttga
      781 cactctatcg ttgatagagt tattttacca ctccctatca gtgatagaga aaagaattga
      841 aaagatctaa agaggagaaa ggatctatgg ataagaaata ctcaataggc ttagctatcg
      901 gcacaaatag cgtcggatgg gcggtgatca ctgatgaata taaggttccg tctaaaaagt
      961 tcaaggttct gggaaataca gaccgccaca gtatcaaaaa aaatcttata ggggctcttt
     1021 tatttgacag tggagagaca gcggaagcga ctcgcctcaa acggacagct cgtagaaggt
     1081 atacacgtcg gaagaatcgt atttgttatc tacaggagat tttttcaaat gagatggcga
     1141 aagtagatga tagtttcttt catcgacttg aagagtcttt tttggtggaa gaagacaaga
     1201 agcatgaacg tcatcctatt tttggaaata tagtagatga agttgcttat catgagaaat
     1261 atccaactat ctatcatctg cgaaaaaaat tggtagattc tactgataaa gcggatttgc
     1321 gcttaatcta tttggcctta gcgcatatga ttaagtttcg tggtcatttt ttgattgagg
     1381 gagatttaaa tcctgataat agtgatgtgg acaaactatt tatccagttg gtacaaacct
     1441 acaatcaatt atttgaagaa aaccctatta acgcaagtgg agtagatgct aaagcgattc
     1501 tttctgcacg attgagtaaa tcaagacgat tagaaaatct cattgctcag ctccccggtg
     1561 agaagaaaaa tggcttattt gggaatctca ttgctttgtc attgggtttg acccctaatt
     1621 ttaaatcaaa ttttgatttg gcagaagatg ctaaattaca gctttcaaaa gatacttacg
     1681 atgatgattt agataattta ttggcgcaaa ttggagatca atatgctgat ttgtttttgg
     1741 cagctaagaa tttatcagat gctattttac tttcagatat cctaagagta aatactgaaa
     1801 taactaaggc tcccctatca gcttcaatga ttaaacgcta cgatgaacat catcaagact
     1861 tgactctttt aaaagcttta gttcgacaac aacttccaga aaagtataaa gaaatctttt
     1921 ttgatcaatc aaaaaacgga tatgcaggtt atattgatgg gggagctagc caagaagaat
     1981 tttataaatt tatcaaacca attttagaaa aaatggatgg tactgaggaa ttattggtga
     2041 aactaaatcg tgaagatttg ctgcgcaagc aacggacctt tgacaacggc tctattcccc
     2101 atcaaattca cttgggtgag ctgcatgcta ttttgagaag acaagaagac ttttatccat
     2161 ttttaaaaga caatcgtgag aagattgaaa aaatcttgac ttttcgtatt ccttattatg
     2221 ttggtccatt ggcgcgtggc aatagtcgtt ttgcatggat gactcggaag tctgaagaaa
     2281 caattacccc atggaatttt gaagaagttg tcgataaagg tgcttcagct caatcattta
     2341 ttgaacgcat gacaaacttt gataaaaatc ttccaaatga aaaagtacta ccaaaacata
     2401 gtttgcttta tgagtatttt acggtttata acgaattgac aaaggtcaaa tatgttactg
     2461 aaggaatgcg aaaaccagca tttctttcag gtgaacagaa gaaagccatt gttgatttac
     2521 tcttcaaaac aaatcgaaaa gtaaccgtta agcaattaaa agaagattat ttcaaaaaaa
     2581 tagaatgttt tgatagtgtt gaaatttcag gagttgaaga tagatttaat gcttcattag
     2641 gtacctacca tgatttgcta aaaattatta aagataaaga ttttttggat aatgaagaaa
     2701 atgaagatat cttagaggat attgttttaa cattgacctt atttgaagat agggagatga
     2761 ttgaggaaag acttaaaaca tatgctcacc tctttgatga taaggtgatg aaacagctta
     2821 aacgtcgccg ttatactggt tggggacgtt tgtctcgaaa attgattaat ggtattaggg
     2881 ataagcaatc tggcaaaaca atattagatt ttttgaaatc agatggtttt gccaatcgca
     2941 attttatgca gctgatccat gatgatagtt tgacatttaa agaagacatt caaaaagcac
     3001 aagtgtctgg acaaggcgat agtttacatg aacatattgc aaatttagct ggtagccctg
     3061 ctattaaaaa aggtatttta cagactgtaa aagttgttga tgaattggtc aaagtaatgg
     3121 ggcggcataa gccagaaaat atcgttattg aaatggcacg tgaaaatcag acaactcaaa
     3181 agggccagaa aaattcgcga gagcgtatga aacgaatcga agaaggtatc aaagaattag
     3241 gaagtcagat tcttaaagag catcctgttg aaaatactca attgcaaaat gaaaagctct
     3301 atctctatta tctccaaaat ggaagagaca tgtatgtgga ccaagaatta gatattaatc
     3361 gtttaagtga ttatgatgtc gatgccattg ttccacaaag tttccttaaa gacgattcaa
     3421 tagacaataa ggtcttaacg cgttctgata aaaatcgtgg taaatcggat aacgttccaa
     3481 gtgaagaagt agtcaaaaag atgaaaaact attggagaca acttctaaac gccaagttaa
     3541 tcactcaacg taagtttgat aatttaacga aagctgaacg tggaggtttg agtgaacttg
     3601 ataaagctgg ttttatcaaa cgccaattgg ttgaaactcg ccaaatcact aagcatgtgg
     3661 cacaaatttt ggatagtcgc atgaatacta aatacgatga aaatgataaa cttattcgag
     3721 aggttaaagt gattacctta aaatctaaat tagtttctga cttccgaaaa gatttccaat
     3781 tctataaagt acgtgagatt aacaattacc atcatgccca tgatgcgtat ctaaatgccg
     3841 tcgttggaac tgctttgatt aagaaatatc caaaacttga atcggagttt gtctatggtg
     3901 attataaagt ttatgatgtt cgtaaaatga ttgctaagtc tgagcaagaa ataggcaaag
     3961 caaccgcaaa atatttcttt tactctaata tcatgaactt cttcaaaaca gaaattacac
     4021 ttgcaaatgg agagattcgc aaacgccctc taatcgaaac taatggggaa actggagaaa
     4081 ttgtctggga taaagggcga gattttgcca cagtgcgcaa agtattgtcc atgccccaag
     4141 tcaatattgt caagaaaaca gaagtacaga caggcggatt ctccaaggag tcaattttac
     4201 caaaaagaaa ttcggacaag cttattgctc gtaaaaaaga ctgggatcca aaaaaatatg
     4261 gtggttttga tagtccaacg gtagcttatt cagtcctagt ggttgctaag gtggaaaaag
     4321 ggaaatcgaa gaagttaaaa tccgttaaag agttactagg gatcacaatt atggaaagaa
     4381 gttcctttga aaaaaatccg attgactttt tagaagctaa aggatataag gaagttaaaa
     4441 aagacttaat cattaaacta cctaaatata gtctttttga gttagaaaac ggtcgtaaac
     4501 ggatgctggc tagtgccgga gaattacaaa aaggaaatga gctggctctg ccaagcaaat
     4561 atgtgaattt tttatattta gctagtcatt atgaaaagtt gaagggtagt ccagaagata
     4621 acgaacaaaa acaattgttt gtggagcagc ataagcatta tttagatgag attattgagc
     4681 aaatcagtga attttctaag cgtgttattt tagcagatgc caatttagat aaagttctta
     4741 gtgcatataa caaacataga gacaaaccaa tacgtgaaca agcagaaaat attattcatt
     4801 tatttacgtt gacgaatctt ggagctcccg ctgcttttaa atattttgat acaacaattg
     4861 atcgtaaacg atatacgtct acaaaagaag ttttagatgc cactcttatc catcaatcca
     4921 tcactggtct ttatgaaaca cgcattgatt tgagtcagct aggaggtgac taatactaga
     4981 gccaattatt gaaggggagc gggaaaccgc tccccttttt ttgtttctgg tctcccctct
     5041 agaagcggcc gcgaattcca gaaatcatcc ttagcgaaag ctaaggattt tttttatctg
     5101 aaattctgcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5161 tagacgtcag gtggcaccat gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5221 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5281 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5341 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5401 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5461 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5521 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5581 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5641 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5701 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5761 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5821 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5881 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5941 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6001 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6061 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6121 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttacgagctc
     6181 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     6241 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcacta gggacagtaa
     6301 gacgggtaag cctgttgatg ataccgctgc cttactgggt gcattagcca gtctgaatga
     6361 cctgtcacgg gataatccga agtggtcaga ctggaaaatc agagggcagg aactgctgaa
     6421 cagcaaaaag tcagatagca ccacatagca gacccgccat aaaacgccct gagaagcccg
     6481 tgacgggctt ttcttgtatt atgggtagtt tccttgcatg aatccataaa aggcgcctgt
     6541 agtgccattt acccccattc actgccagag ccgtgagcgc agcgaactga atgtcacgaa
     6601 aaagacagcg actcaggtgc ctgatggtcg gagacaaaag gaatattcag cgatttgccc
     6661 gagcttgcga gggtgctact taagccttta gggttttaag gtctgttttg tagaggagca
     6721 aacagcgttt gcgacatcct tttgtaatac tgcggaactg actaaagtag tgagttatac
     6781 acagggctgg gatctattct ttttatcttt ttttattctt tctttattct ataaattata
     6841 accacttgaa tataaacaaa aaaaacacac aaaggtctag cggaatttac agagggtcta
     6901 gcagaattta caagttttcc agcaaaggtc tagcagaatt tacagatacc cacaactcaa
     6961 aggaaaagga catgtaatta tcattgacta gcccatctca attggtatag tgattaaaat
     7021 cacctagacc aattgagatg tatgtctgaa ttagttgttt tcaaagcaaa tgaactagcg
     7081 attagtcgct atgacttaac ggagcatgaa accaagctaa ttttatgctg tgtggcacta
     7141 ctcaacccca cgattgaaaa ccctacaagg aaagaacgga cggtatcgtt cacttataac
     7201 caatacgctc agatgatgaa catcagtagg gaaaatgctt atggtgtatt agctaaagca
     7261 accagagagc tgatgacgag aactgtggaa atcaggaatc ctttggttaa aggctttgag
     7321 attttccagt ggacaaacta tgccaagttc tcaagcgaaa aattagaatt agtttttagt
     7381 gaagagatat tgccttatct tttccagtta aaaaaattca taaaatataa tctggaacat
     7441 gttaagtctt ttgaaaacaa atactctatg aggatttatg agtggttatt aaaagaacta
     7501 acacaaaaga aaactcacaa ggcaaatata gagattagcc ttgatgaatt taagttcatg
     7561 ttaatgcttg aaaataacta ccatgagttt aaaaggctta accaatgggt tttgaaacca
     7621 ataagtaaag atttaaacac ttacagcaat atgaaattgg tggttgataa gcgaggccgc
     7681 ccgactgata cgttgatttt ccaagttgaa ctagatagac aaatggatct cgtaaccgaa
     7741 cttgagaaca accagataaa aatgaatggt gacaaaatac caacaaccat tacatcagat
     7801 tcctacctac ataacggact aagaaaaaca ctacacgatg ctttaactgc aaaaattcag
     7861 ctcaccagtt ttgaggcaaa atttttgagt gacatgcaaa gtaagtatga tctcaatggt
     7921 tcgttctcat ggctcacgca aaaacaacga accacactag agaacatact ggctaaatac
     7981 ggaaggatct gaggttctta tggctcttgt atctatcagt gaagcatcaa gactaacaaa
     8041 caaaagtaga acaactgttc accgttacat atcaaaggga aaactgtcca tatgcacaga
     8101 tgaaaacggt gtaaaaaaga tagatacatc agagctttta cgagtttttg gtgcattcaa
     8161 agctgttcac catgaacaga tcgacaatgt aacctcgaga ttaccgcctt tgagtgagct
     8221 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcctgc
     8281 ataacgcgaa gtaatctttt cggttttaaa gaaaaagggc agggtggtga caccttgccc
     8341 ttttttgccg gactgcagcg gccgctacta gtagggagac cagaaacaaa aaaaggccgc
     8401 gttagcggcc ttcaataatt ggctctagta ttattagtag atgccggagt ttgctgcgat
     8461 tgctgaggga atcgggtttc catcttttag gagaccttgc attgccttaa caataagctc
     8521 gcagtcggag ttcgtggcga aaattggaat ggttagttcc atatttaagt acgaacgcca
     8581 ggcgccttta ggcacctcga ctttgatggt gtatttgcga ttctgcgcag agctctgacg
     8641 aacgctacag gttactttgt aagcctgtga acgcgagtta gagctgatcc attcagcgat
     8701 cccgttagcg aagttgcttg gggcgacagt cacgtcgcca gttccgccat tgtcgacgag
     8761 aacgaactga gtaaagttag aagccatttg caaactgagt tgcagcaact ctttttcctg
     8821 ctgcatctga aactcacgta aatccagcgg cagtgttgga agcgaggtgg tttctgaaac
     8881 ggcgatagcg tcttcaggcg gacgccgttt aaagggatca ataatgatgt catcaagcgg
     8941 ataatcgctg gtgccgtggc gatacactga acgttccacc acgtttttca attcacgaat
     9001 atttcccggc caacgataat tcagcaatgt ttctctggcg cgctccgtaa accccgggaa
     9061 cagaggcagc ttgatttccc gacacatctg gatggcaaag tattctgcca tcaacattat
     9121 gtcgctttcg cgctcgcgca gtggtggcag ttgtacaaca tcaaaagcca gtcggtcgag
     9181 caggtcagcg cgaaaagtgc cttcattgac catcgccggg agatcggcat tcgtcgcgca
     9241 taccaaccgc acattcacct gcaatggttg gctgccgcca acgcgctcca gttcaccgta
     9301 ctcaatcacg cgcaataatt tctcctgcac catcatcggt gccgtagcga gttcatcaag
     9361 aaatagcgta ccgccgtcgg cacgttcaaa tctccctgga tgacgttttt gcgcaccggt
     9421 aaacgccccc gcttcgtgac caaacagttc ggaatccagc agattttcat ttaacgccgc
     9481 gcagttaagg gaaataaacg gcccttgcca acgggaggag agataatgca ggcggctggc
     9541 aatcagctct ttaccggtgc cgcgttcgcc gatgatgagc accggtttgt ccagcggtgc
     9601 gagatgcgaa acctgttcca gcacttcgag aaagctgttc gcctcaccaa gtaaattatc
     9661 tttgtattct gccatctagt actttcctgt gtgactctag tagctagcac tatacctagg
     9721 actgagctag ccgtaaactc tagtagggag accagaaaca aaaaaaggcc ccccgttagg
     9781 gaggccttca ataattggct ctagtattat taaactgatg cagcgtagtt ttcgtcgttt
     9841 gcagcaggcc ttttgtacag ttcatccata ccatgcgtga tgcccgctgc ggttacgaac
     9901 tccagcagaa ccatatgatc gcgtttctcg ttcggatctt tagacagaac gctttgcgtg
     9961 ctcagatagt gattgtctgg cagcagaaca ggaccatcac cgattggagt gttttgctgg
    10021 tagtgatcag ccagctgcac gctgccatcc tccacgttgt ggcgaatttt aaaattcgct
    10081 ttaatgccat ttttttgttt atcggcggtg atgtaaacat tgtggctgtt aaaattgtat
    10141 tccagcttat ggcccaggat attgccgtct tctttaaagt caatgccttt cagctcaatg
    10201 cggtttacca gggtatcgcc ttcaaatttc acttccgcac gcgttttgta cgtgccgtca
    10261 tccttaaagg aaatcgtgcg ttcctgcaca tagccttccg gcatggcgga cttgaagaag
    10321 tcatgctgct tcatatggtc cggataacga gcaaagcact gaacaccata agtcagcgtc
    10381 gttaccagag tcggccaagg taccggcagt ttaccagtag tacagatgaa cttcagcgtc
    10441 agtttaccat tagttgcgtc accttcaccc tcgccacgca cggaaaactt atgaccgttg
    10501 acatcaccat ccagttccac cagaataggg acgacaccag tgaacagctc ttcgccttta
    10561 cgcatatggt atttctcctc tttaatctct agttaaacaa aattatttgt agaggctgtt
    10621 tcgtcctcac ggactcatca gaccggaaag cacatccggt gacagctctc tagtatttgc
    10681 gttcaggatt gtcctgctga actgttaata caatttgcgt gccaattttt tatctttttg
    10741 atttataaag atctgattga aga
//
