LOCUS       sequence-257304-       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-LEA1B1 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: pLY53
FEATURES             Location/Qualifiers
     source          1..10491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..29
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(33..76)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    79..100
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(164..1750,1751..1753,1754..4240,4241..4243,
                     4244..4270))
                     /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             4277..4288
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        4306..4361
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    4307..4325
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             4377..5003
                     /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(4453..4472)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             5018..5029
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5069..5103
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             6102..6113
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        6335..6384
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             6417..6428
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6435..7148
                     /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"
     misc_feature    7255..7275
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      7276..7333
                     /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(7357..7374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(7636..8586)
                     /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      8634..8856
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(9348..9442)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(9472..10263,10264..10332))
                     /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     10095..10114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10333..10437)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt
       61 cgctaaggat gatttctgga attcgcggcc gcttctagag gggagaccag aaacaaaaaa
      121 aggggagcgg tttcccgctc cccttcaata attggctcta gtattagtca cctcctagct
      181 gactcaaatc aatgcgtgtt tcataaagac cagtgatgga ttgatggata agagtggcat
      241 ctaaaacttc ttttgtagac gtatatcgtt tacgatcaat tgttgtatca aaatatttaa
      301 aagcagcggg agctccaaga ttcgtcaacg taaataaatg aataatattt tctgcttgtt
      361 cacgtattgg tttgtctcta tgtttgttat atgcactaag aactttatct aaattggcat
      421 ctgctaaaat aacacgctta gaaaattcac tgatttgctc aataatctca tctaaataat
      481 gcttatgctg ctccacaaac aattgttttt gttcgttatc ttctggacta cccttcaact
      541 tttcataatg actagctaaa tataaaaaat tcacatattt gcttggcaga gccagctcat
      601 ttcctttttg taattctccg gcactagcca gcatccgttt acgaccgttt tctaactcaa
      661 aaagactata tttaggtagt ttaatgatta agtctttttt aacttcctta tatcctttag
      721 cttctaaaaa gtcaatcgga tttttttcaa aggaacttct ttccataatt gtgatcccta
      781 gtaactcttt aacggatttt aacttcttcg atttcccttt ttccacctta gcaaccacta
      841 ggactgaata agctaccgtt ggactatcaa aaccaccata tttttttgga tcccagtctt
      901 ttttacgagc aataagcttg tccgaatttc tttttggtaa aattgactcc ttggagaatc
      961 cgcctgtctg tacttctgtt ttcttgacaa tattgacttg gggcatggac aatactttgc
     1021 gcactgtggc aaaatctcgc cctttatccc agacaatttc tccagtttcc ccattagttt
     1081 cgattagagg gcgtttgcga atctctccat ttgcaagtgt aatttctgtt ttgaagaagt
     1141 tcatgatatt agagtaaaag aaatattttg cggttgcttt gcctatttct tgctcagact
     1201 tagcaatcat tttacgaaca tcataaactt tataatcacc atagacaaac tccgattcaa
     1261 gttttggata tttcttaatc aaagcagttc caacgacggc atttagatac gcatcatggg
     1321 catgatggta attgttaatc tcacgtactt tatagaattg gaaatctttt cggaagtcag
     1381 aaactaattt agattttaag gtaatcactt taacctctcg aataagttta tcattttcat
     1441 cgtatttagt attcatgcga ctatccaaaa tttgtgccac atgcttagtg atttggcgag
     1501 tttcaaccaa ttggcgtttg ataaaaccag ctttatcaag ttcactcaaa cctccacgtt
     1561 cagctttcgt taaattatca aacttacgtt gagtgattaa cttggcgttt agaagttgtc
     1621 tccaatagtt tttcatcttt ttgactactt cttcacttgg aacgttatcc gatttaccac
     1681 gatttttatc agaacgcgtt aagaccttat tgtctattga atcgtcttta aggaaacttt
     1741 gtggaacaat ggcatcgaca tcataatcac ttaaacgatt aatatctaat tcttggtcca
     1801 catacatgtc tcttccattt tggagataat agagatagag cttttcattt tgcaattgag
     1861 tattttcaac aggatgctct ttaagaatct gacttcctaa ttctttgata ccttcttcga
     1921 ttcgtttcat acgctctcgc gaatttttct ggcccttttg agttgtctga ttttcacgtg
     1981 ccatttcaat aacgatattt tctggcttat gccgccccat tactttgacc aattcatcaa
     2041 caacttttac agtctgtaaa ataccttttt taatagcagg gctaccagct aaatttgcaa
     2101 tatgttcatg taaactatcg ccttgtccag acacttgtgc tttttgaatg tcttctttaa
     2161 atgtcaaact atcatcatgg atcagctgca taaaattgcg attggcaaaa ccatctgatt
     2221 tcaaaaaatc taatattgtt ttgccagatt gcttatccct aataccatta atcaattttc
     2281 gagacaaacg tccccaacca gtataacggc gacgtttaag ctgtttcatc accttatcat
     2341 caaagaggtg agcatatgtt ttaagtcttt cctcaatcat ctccctatct tcaaataagg
     2401 tcaatgttaa aacaatatcc tctaagatat cttcattttc ttcattatcc aaaaaatctt
     2461 tatctttaat aatttttagc aaatcatggt aggtacctaa tgaagcatta aatctatctt
     2521 caactcctga aatttcaaca ctatcaaaac attctatttt tttgaaataa tcttctttta
     2581 attgcttaac ggttactttt cgatttgttt tgaagagtaa atcaacaatg gctttcttct
     2641 gttcacctga aagaaatgct ggttttcgca ttccttcagt aacatatttg acctttgtca
     2701 attcgttata aaccgtaaaa tactcataaa gcaaactatg ttttggtagt actttttcat
     2761 ttggaagatt tttatcaaag tttgtcatgc gttcaataaa tgattgagct gaagcacctt
     2821 tatcgacaac ttcttcaaaa ttccatgggg taattgtttc ttcagacttc cgagtcatcc
     2881 atgcaaaacg actattgcca cgcgccaatg gaccaacata ataaggaata cgaaaagtca
     2941 agattttttc aatcttctca cgattgtctt ttaaaaatgg ataaaagtct tcttgtcttc
     3001 tcaaaatagc atgcagctca cccaagtgaa tttgatgggg aatagagccg ttgtcaaagg
     3061 tccgttgctt gcgcagcaaa tcttcacgat ttagtttcac caataattcc tcagtaccat
     3121 ccattttttc taaaattggt ttgataaatt tataaaattc ttcttggcta gctcccccat
     3181 caatataacc tgcatatccg ttttttgatt gatcaaaaaa gatttcttta tacttttctg
     3241 gaagttgttg tcgaactaaa gcttttaaaa gagtcaagtc ttgatgatgt tcatcgtagc
     3301 gtttaatcat tgaagctgat aggggagcct tagttatttc agtatttact cttaggatat
     3361 ctgaaagtaa aatagcatct gataaattct tagctgccaa aaacaaatca gcatattgat
     3421 ctccaatttg cgccaataaa ttatctaaat catcatcgta agtatctttt gaaagctgta
     3481 atttagcatc ttctgccaaa tcaaaatttg atttaaaatt aggggtcaaa cccaatgaca
     3541 aagcaatgag attcccaaat aagccatttt tcttctcacc ggggagctga gcaatgagat
     3601 tttctaatcg tcttgattta ctcaatcgtg cagaaagaat cgctttagca tctactccac
     3661 ttgcgttaat agggttttct tcaaataatt gattgtaggt ttgtaccaac tggataaata
     3721 gtttgtccac atcactatta tcaggattta aatctccctc aatcaaaaaa tgaccacgaa
     3781 acttaatcat atgcgctaag gccaaataga ttaagcgcaa atccgcttta tcagtagaat
     3841 ctaccaattt ttttcgcaga tgatagatag ttggatattt ctcatgataa gcaacttcat
     3901 ctactatatt tccaaaaata ggatgacgtt catgcttctt gtcttcttcc accaaaaaag
     3961 actcttcaag tcgatgaaag aaactatcat ctactttcgc catctcattt gaaaaaatct
     4021 cctgtagata acaaatacga ttcttccgac gtgtatacct tctacgagct gtccgtttga
     4081 ggcgagtcgc ttccgctgtc tctccactgt caaataaaag agcccctata agattttttt
     4141 tgatactgtg gcggtctgta tttcccagaa ccttgaactt tttagacgga accttatatt
     4201 catcagtgat caccgcccat ccgacgctat ttgtgccgat agctaagcct attgagtatt
     4261 tcttatccat agatcctttc tcctctttag atcttttcaa ttcttttctc tatcactgat
     4321 agggagtggt aaaataactc tatcaacgat agagtgtcaa caaaaattag gaattaatga
     4381 tgtcaagatt agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg
     4441 gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat
     4501 tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag
     4561 ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta
     4621 ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca aaagtacatt
     4681 taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat
     4741 gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg gggcatttta
     4801 ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac
     4861 ctactactga tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag
     4921 gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta gaaaaacaac
     4981 ttaaatgtga aagtgggtct taatactaga gccaattatt gaaggccgct aacgcggcct
     5041 ttttttgttt ctggtctccc tactagagtt tacggctagc tcagtcctag gtatagtgct
     5101 agctactaga gtcacacagg aaagtactag atggcagaat acaaagataa tttacttggt
     5161 gaggcgaaca gctttctcga agtgctggaa caggtttcgc atctcgcacc gctggacaaa
     5221 ccggtgctca tcatcggcga acgcggcacc ggtaaagagc tgattgccag ccgcctgcat
     5281 tatctctcct cccgttggca agggccgttt atttccctta actgcgcggc gttaaatgaa
     5341 aatctgctgg attccgaact gtttggtcac gaagcggggg cgtttaccgg tgcgcaaaaa
     5401 cgtcatccag ggagatttga acgtgccgac ggcggtacgc tatttcttga tgaactcgct
     5461 acggcaccga tgatggtgca ggagaaatta ttgcgcgtga ttgagtacgg tgaactggag
     5521 cgcgttggcg gcagccaacc attgcaggtg aatgtgcggt tggtatgcgc gacgaatgcc
     5581 gatctcccgg cgatggtcaa tgaaggcact tttcgcgctg acctgctcga ccgactggct
     5641 tttgatgttg tacaactgcc accactgcgc gagcgcgaaa gcgacataat gttgatggca
     5701 gaatactttg ccatccagat gtgtcgggaa atcaagctgc ctctgttccc ggggtttacg
     5761 gagcgcgcca gagaaacatt gctgaattat cgttggccgg gaaatattcg tgaattgaaa
     5821 aacgtggtgg aacgttcagt gtatcgccac ggcaccagcg attatccgct tgatgacatc
     5881 attattgatc cctttaaacg gcgtccgcct gaagacgcta tcgccgtttc agaaaccacc
     5941 tcgcttccaa cactgccgct ggatttacgt gagtttcaga tgcagcagga aaaagagttg
     6001 ctgcaactca gtttgcaaat gaatgcacgc acacgccgcc gcgaacgtcg cgcagagaaa
     6061 caggctcaat ggaaagcagc aaattaatac tagagccaat tattgaaggc cgctaacgcg
     6121 gccttttttt gtttctggtc tccctactag aagttatccc gacgcgccgt ccggtctgag
     6181 gtctggaaga tcggttagtg taattcgcta actcatcctg gcatgttgct gttgattctt
     6241 caatcagatc tttataaatc aaaaagataa aaaattggca cgcaaattgt attaacagtt
     6301 cagcaggaca atcctgaacg caaatactag agagctgtca ccggatgtgc tttccggtct
     6361 gatgagtccg tgaggacgaa acagcctcta caaataattt tgtttaacta gagattaaag
     6421 aggagaaata ccatatgcgt aaaggcgaag agctgttcac tggtgtcgtc cctattctgg
     6481 tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg
     6541 acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggtac
     6601 cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct cgttatccgg
     6661 accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac
     6721 gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag
     6781 gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagaa gacggcaata
     6841 tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc accgccgata
     6901 aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg
     6961 tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc
     7021 cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg
     7081 atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac
     7141 tgtacaaaag gcctgctgca aacgacgaaa actacgctgc atcagtttaa taatactaga
     7201 gccaattatt gaaggcctcc ctaacggggg gccttttttt gtttctggtc tccctactag
     7261 tagcggccgc tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc ctttttcttt
     7321 aaaaccgaaa agattacttc gcgttatgca ggcttcctcg ctcactgact cgctgcgctc
     7381 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatct cgaggttaca
     7441 ttgtcgatct gttcatggtg aacagctttg aatgcaccaa aaactcgtaa aagctctgat
     7501 gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc ctttgatatg
     7561 taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca
     7621 agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg
     7681 ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt gcatgtcact
     7741 caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt
     7801 ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc
     7861 attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc
     7921 aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt
     7981 gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa
     8041 ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat
     8101 atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat
     8161 agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa
     8221 ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa
     8281 cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc
     8341 cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct
     8401 actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc gttctttcct
     8461 tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc
     8521 atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc
     8581 agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag
     8641 tcaatgataa ttacatgtcc ttttcctttg agttgtgggt atctgtaaat tctgctagac
     8701 ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg
     8761 tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa
     8821 gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta
     8881 ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa
     8941 aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga
     9001 ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc
     9061 tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac
     9121 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc
     9181 tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct
     9241 gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca
     9301 gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt ctcaccaata
     9361 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
     9421 actggatcta tcaacaggag tccaagcgag ctcgtaaact tggtctgaca gttaccaatg
     9481 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     9541 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     9601 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     9661 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     9721 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     9781 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     9841 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     9901 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     9961 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    10021 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    10081 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    10141 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    10201 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    10261 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    10321 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    10381 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    10441 atttcccatg gtgccacctg acgtctaaga aaccattatt atcatgacat t
//
