LOCUS       sequence-257327-       10491 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-LEA5B5 with 
            sfgfp::ASV)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10491)
  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 10491)
  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: pLY57
FEATURES             Location/Qualifiers
     source          1..10491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    365..385
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      386..443
                     /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."
     primer_bind     complement(467..484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(746..1696)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1744..1966
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2458..2552)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2582..3373,3374..3442))
                     /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     3205..3224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3443..3547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3612..3630
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3634..3677)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3680..3701
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(3765..5351,5352..5354,5355..7841,7842..7844,
                     7845..7871))
                     /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"
     RBS             7878..7889
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        7907..7962
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7908..7926
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             7978..8604
                     /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     complement(8054..8073)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             8619..8630
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        8670..8704
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             9703..9714
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        9936..9985
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             10018..10029
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(10036..10491,1..258)
                     /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 accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag
       61 gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct
      121 gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac
      181 gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt
      241 atggatgaac tgtacaaaag gcctgctgca aacgacgaaa actacgctgc atcagtttaa
      301 taatactaga gccaattatt gaaggcctcc ctaacggggg gccttttttt gtttctggtc
      361 tccctactag tagcggccgc tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc
      421 ctttttcttt aaaaccgaaa agattacttc gcgttatgca ggcttcctcg ctcactgact
      481 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatct
      541 cgaggttaca ttgtcgatct gttcatggtg aacagctttg aatgcaccaa aaactcgtaa
      601 aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc
      661 ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact
      721 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta
      781 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt
      841 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat
      901 cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta
      961 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc
     1021 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca
     1081 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa
     1141 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc
     1201 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat
     1261 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga
     1321 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt
     1381 cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat
     1441 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag
     1501 cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc
     1561 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
     1621 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
     1681 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
     1741 gatgggctag tcaatgataa ttacatgtcc ttttcctttg agttgtgggt atctgtaaat
     1801 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
     1861 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
     1921 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
     1981 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
     2041 aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt
     2101 ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg
     2161 ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat
     2221 gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg
     2281 gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat
     2341 tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc
     2401 cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt
     2461 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     2521 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgtaaact tggtctgaca
     2581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2881 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3181 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3541 ttccgcgcac atttcccatg gtgccacctg acgtctaaga aaccattatt atcatgacat
     3601 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
     3661 tcgctaagga tgatttctgg aattcgcggc cgcttctaga ggggagacca gaaacaaaaa
     3721 aaggggagcg gtttcccgct ccccttcaat aattggctct agtattagtc acctcctagc
     3781 tgactcaaat caatgcgtgt ttcataaaga ccagtgatgg attgatggat aagagtggca
     3841 tctaaaactt cttttgtaga cgtatatcgt ttacgatcaa ttgttgtatc aaaatattta
     3901 aaagcagcgg gagctccaag attcgtcaac gtaaataaat gaataatatt ttctgcttgt
     3961 tcacgtattg gtttgtctct atgtttgtta tatgcactaa gaactttatc taaattggca
     4021 tctgctaaaa taacacgctt agaaaattca ctgatttgct caataatctc atctaaataa
     4081 tgcttatgct gctccacaaa caattgtttt tgttcgttat cttctggact acccttcaac
     4141 ttttcataat gactagctaa atataaaaaa ttcacatatt tgcttggcag agccagctca
     4201 tttccttttt gtaattctcc ggcactagcc agcatccgtt tacgaccgtt ttctaactca
     4261 aaaagactat atttaggtag tttaatgatt aagtcttttt taacttcctt atatccttta
     4321 gcttctaaaa agtcaatcgg atttttttca aaggaacttc tttccataat tgtgatccct
     4381 agtaactctt taacggattt taacttcttc gatttccctt tttccacctt agcaaccact
     4441 aggactgaat aagctaccgt tggactatca aaaccaccat atttttttgg atcccagtct
     4501 tttttacgag caataagctt gtccgaattt ctttttggta aaattgactc cttggagaat
     4561 ccgcctgtct gtacttctgt tttcttgaca atattgactt ggggcatgga caatactttg
     4621 cgcactgtgg caaaatctcg ccctttatcc cagacaattt ctccagtttc cccattagtt
     4681 tcgattagag ggcgtttgcg aatctctcca tttgcaagtg taatttctgt tttgaagaag
     4741 ttcatgatat tagagtaaaa gaaatatttt gcggttgctt tgcctatttc ttgctcagac
     4801 ttagcaatca ttttacgaac atcataaact ttataatcac catagacaaa ctccgattca
     4861 agttttggat atttcttaat caaagcagtt ccaacgacgg catttagata cgcatcatgg
     4921 gcatgatggt aattgttaat ctcacgtact ttatagaatt ggaaatcttt tcggaagtca
     4981 gaaactaatt tagattttaa ggtaatcact ttaacctctc gaataagttt atcattttca
     5041 tcgtatttag tattcatgcg actatccaaa atttgtgcca catgcttagt gatttggcga
     5101 gtttcaacca attggcgttt gataaaacca gctttatcaa gttcactcaa acctccacgt
     5161 tcagctttcg ttaaattatc aaacttacgt tgagtgatta acttggcgtt tagaagttgt
     5221 ctccaatagt ttttcatctt tttgactact tcttcacttg gaacgttatc cgatttacca
     5281 cgatttttat cagaacgcgt taagacctta ttgtctattg aatcgtcttt aaggaaactt
     5341 tgtggaacaa tggcatcgac atcataatca cttaaacgat taatatctaa ttcttggtcc
     5401 acatacatgt ctcttccatt ttggagataa tagagataga gcttttcatt ttgcaattga
     5461 gtattttcaa caggatgctc tttaagaatc tgacttccta attctttgat accttcttcg
     5521 attcgtttca tacgctctcg cgaatttttc tggccctttt gagttgtctg attttcacgt
     5581 gccatttcaa taacgatatt ttctggctta tgccgcccca ttactttgac caattcatca
     5641 acaactttta cagtctgtaa aatacctttt ttaatagcag ggctaccagc taaatttgca
     5701 atatgttcat gtaaactatc gccttgtcca gacacttgtg ctttttgaat gtcttcttta
     5761 aatgtcaaac tatcatcatg gatcagctgc ataaaattgc gattggcaaa accatctgat
     5821 ttcaaaaaat ctaatattgt tttgccagat tgcttatccc taataccatt aatcaatttt
     5881 cgagacaaac gtccccaacc agtataacgg cgacgtttaa gctgtttcat caccttatca
     5941 tcaaagaggt gagcatatgt tttaagtctt tcctcaatca tctccctatc ttcaaataag
     6001 gtcaatgtta aaacaatatc ctctaagata tcttcatttt cttcattatc caaaaaatct
     6061 ttatctttaa taatttttag caaatcatgg taggtaccta atgaagcatt aaatctatct
     6121 tcaactcctg aaatttcaac actatcaaaa cattctattt ttttgaaata atcttctttt
     6181 aattgcttaa cggttacttt tcgatttgtt ttgaagagta aatcaacaat ggctttcttc
     6241 tgttcacctg aaagaaatgc tggttttcgc attccttcag taacatattt gacctttgtc
     6301 aattcgttat aaaccgtaaa atactcataa agcaaactat gttttggtag tactttttca
     6361 tttggaagat ttttatcaaa gtttgtcatg cgttcaataa atgattgagc tgaagcacct
     6421 ttatcgacaa cttcttcaaa attccatggg gtaattgttt cttcagactt ccgagtcatc
     6481 catgcaaaac gactattgcc acgcgccaat ggaccaacat aataaggaat acgaaaagtc
     6541 aagatttttt caatcttctc acgattgtct tttaaaaatg gataaaagtc ttcttgtctt
     6601 ctcaaaatag catgcagctc acccaagtga atttgatggg gaatagagcc gttgtcaaag
     6661 gtccgttgct tgcgcagcaa atcttcacga tttagtttca ccaataattc ctcagtacca
     6721 tccatttttt ctaaaattgg tttgataaat ttataaaatt cttcttggct agctccccca
     6781 tcaatataac ctgcatatcc gttttttgat tgatcaaaaa agatttcttt atacttttct
     6841 ggaagttgtt gtcgaactaa agcttttaaa agagtcaagt cttgatgatg ttcatcgtag
     6901 cgtttaatca ttgaagctga taggggagcc ttagttattt cagtatttac tcttaggata
     6961 tctgaaagta aaatagcatc tgataaattc ttagctgcca aaaacaaatc agcatattga
     7021 tctccaattt gcgccaataa attatctaaa tcatcatcgt aagtatcttt tgaaagctgt
     7081 aatttagcat cttctgccaa atcaaaattt gatttaaaat taggggtcaa acccaatgac
     7141 aaagcaatga gattcccaaa taagccattt ttcttctcac cggggagctg agcaatgaga
     7201 ttttctaatc gtcttgattt actcaatcgt gcagaaagaa tcgctttagc atctactcca
     7261 cttgcgttaa tagggttttc ttcaaataat tgattgtagg tttgtaccaa ctggataaat
     7321 agtttgtcca catcactatt atcaggattt aaatctccct caatcaaaaa atgaccacga
     7381 aacttaatca tatgcgctaa ggccaaatag attaagcgca aatccgcttt atcagtagaa
     7441 tctaccaatt tttttcgcag atgatagata gttggatatt tctcatgata agcaacttca
     7501 tctactatat ttccaaaaat aggatgacgt tcatgcttct tgtcttcttc caccaaaaaa
     7561 gactcttcaa gtcgatgaaa gaaactatca tctactttcg ccatctcatt tgaaaaaatc
     7621 tcctgtagat aacaaatacg attcttccga cgtgtatacc ttctacgagc tgtccgtttg
     7681 aggcgagtcg cttccgctgt ctctccactg tcaaataaaa gagcccctat aagatttttt
     7741 ttgatactgt ggcggtctgt atttcccaga accttgaact ttttagacgg aaccttatat
     7801 tcatcagtga tcaccgccca tccgacgcta tttgtgccga tagctaagcc tattgagtat
     7861 ttcttatcca tagatccttt ctcctcttta gatcttttca attcttttct ctatcactga
     7921 tagggagtgg taaaataact ctatcaacga tagagtgtca acaaaaatta ggaattaatg
     7981 atgtcaagat tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc
     8041 ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga gcagcctaca
     8101 ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat tgagatgtta
     8161 gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt
     8221 aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat
     8281 ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta
     8341 tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt ggggcatttt
     8401 actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca
     8461 cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa
     8521 ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt agaaaaacaa
     8581 cttaaatgtg aaagtgggtc ttaatactag agccaattat tgaaggccgc taacgcggcc
     8641 tttttttgtt tctggtctcc ctactagagt ttacggctag ctcagtccta ggtatagtgc
     8701 tagctactag agtcacacag gaaagtacta gatggcagaa tacaaagata atttacttgg
     8761 tgaggcgaac agctttctcg aagtgctgga acaggtttcg catctcgcac cgctggacaa
     8821 accggtgctc atcatcggcg aacgcggcac cggtaaagag ctgattgcca gccgcctgca
     8881 ttatctctcc tcccgttggc aagggccgtt tatttccctt aactgcgcgg cgttaaatga
     8941 aaatctgctg gattccgaac tgtttggtca cgaagcgggg gcgtttaccg gtgcgcaaaa
     9001 acgtcatcca gggagatttg aacgtgccga cggcggtacg ctatttcttg atgaactcgc
     9061 tacggcaccg atgatggtgc aggagaaatt attgcgcgtg attgagtacg gtgaactgga
     9121 gcgcgttggc ggcagccaac cattgcaggt gaatgtgcgg ttggtatgcg cgacgaatgc
     9181 cgatctcccg gcgatggtca atgaaggcac ttttcgcgct gacctgctcg accgactggc
     9241 ttttgatgtt gtacaactgc caccactgcg cgagcgcgaa agcgacataa tgttgatggc
     9301 agaatacttt gccatccaga tgtgtcggga aatcaagctg cctctgttcc cggggtttac
     9361 ggagcgcgcc agagaaacat tgctgaatta tcgttggccg ggaaatattc gtgaattgaa
     9421 aaacgtggtg gaacgttcag tgtatcgcca cggcaccagc gattatccgc ttgatgacat
     9481 cattattgat ccctttaaac ggcgtccgcc tgaagacgct atcgccgttt cagaaaccac
     9541 ctcgcttcca acactgccgc tggatttacg tgagtttcag atgcagcagg aaaaagagtt
     9601 gctgcaactc agtttgcaaa tgaatgcacg cacacgccgc cgcgaacgtc gcgcagagaa
     9661 acaggctcaa tggaaagcag caaattaata ctagagccaa ttattgaagg ccgctaacgc
     9721 ggcctttttt tgtttctggt ctccctacta gaagttatat gttggtaact accggggatg
     9781 aatatataag tgcggttagt gtaattcgct aactcatcct ggcatgttgc tgttgattct
     9841 tcaatcagat ctttataaat caaaaagata aaaaattggc acgcaaattg tattaacagt
     9901 tcagcaggac aatcctgaac gcaaatacta gagagctgtc accggatgtg ctttccggtc
     9961 tgatgagtcc gtgaggacga aacagcctct acaaataatt ttgtttaact agagattaaa
    10021 gaggagaaat accatatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg
    10081 gtggaactgg atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt
    10141 gacgcaacta atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggta
    10201 ccttggccga ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg
    10261 gaccatatga agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa
    10321 cgcacgattt cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa
    10381 ggcgataccc tggtaaaccg cattgagctg aaaggcattg actttaaaga agacggcaat
    10441 atcctgggcc ataagctgga atacaatttt aacagccaca atgtttacat c
//
