LOCUS       sequence-257307-       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-LEA2B2 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: pLY54
FEATURES             Location/Qualifiers
     source          1..10491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             464..475
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(508..557)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             complement(779..790)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(1789..1823)
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             complement(1863..1874)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(1889..2515)
                     /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     2420..2439
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        2531..2586
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    2567..2585
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2604..2615
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2622..2648,2649..2651,2652..5138,5139..5141,
                     5142..6728)
                     /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    6792..6813
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      6816..6859
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(6863..6881)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6946..7050
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7051..7119,7120..7911)
                     /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(7269..7288)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7941..8035
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      8527..8749
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             8797..9747
                     /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     10009..10026
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(10050..10107)
                     /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    10108..10128
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(join(10235..10491,1..457))
                     /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 tgatgtaaac attgtggctg ttaaaattgt attccagctt atggcccagg atattgccgt
       61 cttctttaaa gtcaatgcct ttcagctcaa tgcggtttac cagggtatcg ccttcaaatt
      121 tcacttccgc acgcgttttg tacgtgccgt catccttaaa ggaaatcgtg cgttcctgca
      181 catagccttc cggcatggcg gacttgaaga agtcatgctg cttcatatgg tccggataac
      241 gagcaaagca ctgaacacca taagtcagcg tcgttaccag agtcggccaa ggtaccggca
      301 gtttaccagt agtacagatg aacttcagcg tcagtttacc attagttgcg tcaccttcac
      361 cctcgccacg cacggaaaac ttatgaccgt tgacatcacc atccagttcc accagaatag
      421 ggacgacacc agtgaacagc tcttcgcctt tacgcatatg gtatttctcc tctttaatct
      481 ctagttaaac aaaattattt gtagaggctg tttcgtcctc acggactcat cagaccggaa
      541 agcacatccg gtgacagctc tctagtattt gcgttcagga ttgtcctgct gaactgttaa
      601 tacaatttgc gtgccaattt tttatctttt tgatttataa agatctgatt gaagaatcaa
      661 cagcaacatg ccaggatgag ttagcgaatt acactaaccg caaaacaacg tctcaaaccg
      721 gaccgaacta tataataact tctagtaggg agaccagaaa caaaaaaagg ccgcgttagc
      781 ggccttcaat aattggctct agtattaatt tgctgctttc cattgagcct gtttctctgc
      841 gcgacgttcg cggcggcgtg tgcgtgcatt catttgcaaa ctgagttgca gcaactcttt
      901 ttcctgctgc atctgaaact cacgtaaatc cagcggcagt gttggaagcg aggtggtttc
      961 tgaaacggcg atagcgtctt caggcggacg ccgtttaaag ggatcaataa tgatgtcatc
     1021 aagcggataa tcgctggtgc cgtggcgata cactgaacgt tccaccacgt ttttcaattc
     1081 acgaatattt cccggccaac gataattcag caatgtttct ctggcgcgct ccgtaaaccc
     1141 cgggaacaga ggcagcttga tttcccgaca catctggatg gcaaagtatt ctgccatcaa
     1201 cattatgtcg ctttcgcgct cgcgcagtgg tggcagttgt acaacatcaa aagccagtcg
     1261 gtcgagcagg tcagcgcgaa aagtgccttc attgaccatc gccgggagat cggcattcgt
     1321 cgcgcatacc aaccgcacat tcacctgcaa tggttggctg ccgccaacgc gctccagttc
     1381 accgtactca atcacgcgca ataatttctc ctgcaccatc atcggtgccg tagcgagttc
     1441 atcaagaaat agcgtaccgc cgtcggcacg ttcaaatctc cctggatgac gtttttgcgc
     1501 accggtaaac gcccccgctt cgtgaccaaa cagttcggaa tccagcagat tttcatttaa
     1561 cgccgcgcag ttaagggaaa taaacggccc ttgccaacgg gaggagagat aatgcaggcg
     1621 gctggcaatc agctctttac cggtgccgcg ttcgccgatg atgagcaccg gtttgtccag
     1681 cggtgcgaga tgcgaaacct gttccagcac ttcgagaaag ctgttcgcct caccaagtaa
     1741 attatctttg tattctgcca tctagtactt tcctgtgtga ctctagtagc tagcactata
     1801 cctaggactg agctagccgt aaactctagt agggagacca gaaacaaaaa aaggccgcgt
     1861 tagcggcctt caataattgg ctctagtatt aagacccact ttcacattta agttgttttt
     1921 ctaatccgca tatgatcaat tcaaggccga ataagaaggc tggctctgca ccttggtgat
     1981 caaataattc gatagcttgt cgtaataatg gcggcatact atcagtagta ggtgtttccc
     2041 tttcttcttt agcgacttga tgctcttgat cttccaatac gcaacctaaa gtaaaatgcc
     2101 ccacagcgct gagtgcatat aatgcattct ctagtgaaaa accttgttgg cataaaaagg
     2161 ctaattgatt ttcgagagtt tcatactgtt tttctgtagg ccgtgtacct aaatgtactt
     2221 ttgctccatc gcgatgactt agtaaagcac atctaaaact tttagcgtta ttacgtaaaa
     2281 aatcttgcca gctttcccct tctaaagggc aaaagtgagt atggtgccta tctaacatct
     2341 caatggctaa ggcgtcgagc aaagcccgct tattttttac atgccaatac aatgtaggct
     2401 gctctacacc tagcttctgg gcgagtttac gggttgttaa accttcgatt ccgacctcat
     2461 taagcagctc taatgcgctg ttaatcactt tacttttatc taatcttgac atcattaatt
     2521 cctaattttt gttgacactc tatcgttgat agagttattt taccactccc tatcagtgat
     2581 agagaaaaga attgaaaaga tctaaagagg agaaaggatc tatggataag aaatactcaa
     2641 taggcttagc tatcggcaca aatagcgtcg gatgggcggt gatcactgat gaatataagg
     2701 ttccgtctaa aaagttcaag gttctgggaa atacagaccg ccacagtatc aaaaaaaatc
     2761 ttataggggc tcttttattt gacagtggag agacagcgga agcgactcgc ctcaaacgga
     2821 cagctcgtag aaggtataca cgtcggaaga atcgtatttg ttatctacag gagatttttt
     2881 caaatgagat ggcgaaagta gatgatagtt tctttcatcg acttgaagag tcttttttgg
     2941 tggaagaaga caagaagcat gaacgtcatc ctatttttgg aaatatagta gatgaagttg
     3001 cttatcatga gaaatatcca actatctatc atctgcgaaa aaaattggta gattctactg
     3061 ataaagcgga tttgcgctta atctatttgg ccttagcgca tatgattaag tttcgtggtc
     3121 attttttgat tgagggagat ttaaatcctg ataatagtga tgtggacaaa ctatttatcc
     3181 agttggtaca aacctacaat caattatttg aagaaaaccc tattaacgca agtggagtag
     3241 atgctaaagc gattctttct gcacgattga gtaaatcaag acgattagaa aatctcattg
     3301 ctcagctccc cggtgagaag aaaaatggct tatttgggaa tctcattgct ttgtcattgg
     3361 gtttgacccc taattttaaa tcaaattttg atttggcaga agatgctaaa ttacagcttt
     3421 caaaagatac ttacgatgat gatttagata atttattggc gcaaattgga gatcaatatg
     3481 ctgatttgtt tttggcagct aagaatttat cagatgctat tttactttca gatatcctaa
     3541 gagtaaatac tgaaataact aaggctcccc tatcagcttc aatgattaaa cgctacgatg
     3601 aacatcatca agacttgact cttttaaaag ctttagttcg acaacaactt ccagaaaagt
     3661 ataaagaaat cttttttgat caatcaaaaa acggatatgc aggttatatt gatgggggag
     3721 ctagccaaga agaattttat aaatttatca aaccaatttt agaaaaaatg gatggtactg
     3781 aggaattatt ggtgaaacta aatcgtgaag atttgctgcg caagcaacgg acctttgaca
     3841 acggctctat tccccatcaa attcacttgg gtgagctgca tgctattttg agaagacaag
     3901 aagactttta tccattttta aaagacaatc gtgagaagat tgaaaaaatc ttgacttttc
     3961 gtattcctta ttatgttggt ccattggcgc gtggcaatag tcgttttgca tggatgactc
     4021 ggaagtctga agaaacaatt accccatgga attttgaaga agttgtcgat aaaggtgctt
     4081 cagctcaatc atttattgaa cgcatgacaa actttgataa aaatcttcca aatgaaaaag
     4141 tactaccaaa acatagtttg ctttatgagt attttacggt ttataacgaa ttgacaaagg
     4201 tcaaatatgt tactgaagga atgcgaaaac cagcatttct ttcaggtgaa cagaagaaag
     4261 ccattgttga tttactcttc aaaacaaatc gaaaagtaac cgttaagcaa ttaaaagaag
     4321 attatttcaa aaaaatagaa tgttttgata gtgttgaaat ttcaggagtt gaagatagat
     4381 ttaatgcttc attaggtacc taccatgatt tgctaaaaat tattaaagat aaagattttt
     4441 tggataatga agaaaatgaa gatatcttag aggatattgt tttaacattg accttatttg
     4501 aagataggga gatgattgag gaaagactta aaacatatgc tcacctcttt gatgataagg
     4561 tgatgaaaca gcttaaacgt cgccgttata ctggttgggg acgtttgtct cgaaaattga
     4621 ttaatggtat tagggataag caatctggca aaacaatatt agattttttg aaatcagatg
     4681 gttttgccaa tcgcaatttt atgcagctga tccatgatga tagtttgaca tttaaagaag
     4741 acattcaaaa agcacaagtg tctggacaag gcgatagttt acatgaacat attgcaaatt
     4801 tagctggtag ccctgctatt aaaaaaggta ttttacagac tgtaaaagtt gttgatgaat
     4861 tggtcaaagt aatggggcgg cataagccag aaaatatcgt tattgaaatg gcacgtgaaa
     4921 atcagacaac tcaaaagggc cagaaaaatt cgcgagagcg tatgaaacga atcgaagaag
     4981 gtatcaaaga attaggaagt cagattctta aagagcatcc tgttgaaaat actcaattgc
     5041 aaaatgaaaa gctctatctc tattatctcc aaaatggaag agacatgtat gtggaccaag
     5101 aattagatat taatcgttta agtgattatg atgtcgatgc cattgttcca caaagtttcc
     5161 ttaaagacga ttcaatagac aataaggtct taacgcgttc tgataaaaat cgtggtaaat
     5221 cggataacgt tccaagtgaa gaagtagtca aaaagatgaa aaactattgg agacaacttc
     5281 taaacgccaa gttaatcact caacgtaagt ttgataattt aacgaaagct gaacgtggag
     5341 gtttgagtga acttgataaa gctggtttta tcaaacgcca attggttgaa actcgccaaa
     5401 tcactaagca tgtggcacaa attttggata gtcgcatgaa tactaaatac gatgaaaatg
     5461 ataaacttat tcgagaggtt aaagtgatta ccttaaaatc taaattagtt tctgacttcc
     5521 gaaaagattt ccaattctat aaagtacgtg agattaacaa ttaccatcat gcccatgatg
     5581 cgtatctaaa tgccgtcgtt ggaactgctt tgattaagaa atatccaaaa cttgaatcgg
     5641 agtttgtcta tggtgattat aaagtttatg atgttcgtaa aatgattgct aagtctgagc
     5701 aagaaatagg caaagcaacc gcaaaatatt tcttttactc taatatcatg aacttcttca
     5761 aaacagaaat tacacttgca aatggagaga ttcgcaaacg ccctctaatc gaaactaatg
     5821 gggaaactgg agaaattgtc tgggataaag ggcgagattt tgccacagtg cgcaaagtat
     5881 tgtccatgcc ccaagtcaat attgtcaaga aaacagaagt acagacaggc ggattctcca
     5941 aggagtcaat tttaccaaaa agaaattcgg acaagcttat tgctcgtaaa aaagactggg
     6001 atccaaaaaa atatggtggt tttgatagtc caacggtagc ttattcagtc ctagtggttg
     6061 ctaaggtgga aaaagggaaa tcgaagaagt taaaatccgt taaagagtta ctagggatca
     6121 caattatgga aagaagttcc tttgaaaaaa atccgattga ctttttagaa gctaaaggat
     6181 ataaggaagt taaaaaagac ttaatcatta aactacctaa atatagtctt tttgagttag
     6241 aaaacggtcg taaacggatg ctggctagtg ccggagaatt acaaaaagga aatgagctgg
     6301 ctctgccaag caaatatgtg aattttttat atttagctag tcattatgaa aagttgaagg
     6361 gtagtccaga agataacgaa caaaaacaat tgtttgtgga gcagcataag cattatttag
     6421 atgagattat tgagcaaatc agtgaatttt ctaagcgtgt tattttagca gatgccaatt
     6481 tagataaagt tcttagtgca tataacaaac atagagacaa accaatacgt gaacaagcag
     6541 aaaatattat tcatttattt acgttgacga atcttggagc tcccgctgct tttaaatatt
     6601 ttgatacaac aattgatcgt aaacgatata cgtctacaaa agaagtttta gatgccactc
     6661 ttatccatca atccatcact ggtctttatg aaacacgcat tgatttgagt cagctaggag
     6721 gtgactaata ctagagccaa ttattgaagg ggagcgggaa accgctcccc tttttttgtt
     6781 tctggtctcc cctctagaag cggccgcgaa ttccagaaat catccttagc gaaagctaag
     6841 gatttttttt atctgaaatt ctgcctcgtg atacgcctat ttttataggt taatgtcatg
     6901 ataataatgg tttcttagac gtcaggtggc accatgggaa atgtgcgcgg aacccctatt
     6961 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     7021 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     7081 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     7141 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     7201 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     7261 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     7321 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     7381 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     7441 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     7501 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7561 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     7621 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7681 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7741 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7801 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7861 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7921 caagtttacg agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
     7981 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
     8041 cactagggac agtaagacgg gtaagcctgt tgatgatacc gctgccttac tgggtgcatt
     8101 agccagtctg aatgacctgt cacgggataa tccgaagtgg tcagactgga aaatcagagg
     8161 gcaggaactg ctgaacagca aaaagtcaga tagcaccaca tagcagaccc gccataaaac
     8221 gccctgagaa gcccgtgacg ggcttttctt gtattatggg tagtttcctt gcatgaatcc
     8281 ataaaaggcg cctgtagtgc catttacccc cattcactgc cagagccgtg agcgcagcga
     8341 actgaatgtc acgaaaaaga cagcgactca ggtgcctgat ggtcggagac aaaaggaata
     8401 ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc ctttagggtt ttaaggtctg
     8461 ttttgtagag gagcaaacag cgtttgcgac atccttttgt aatactgcgg aactgactaa
     8521 agtagtgagt tatacacagg gctgggatct attcttttta tcttttttta ttctttcttt
     8581 attctataaa ttataaccac ttgaatataa acaaaaaaaa cacacaaagg tctagcggaa
     8641 tttacagagg gtctagcaga atttacaagt tttccagcaa aggtctagca gaatttacag
     8701 atacccacaa ctcaaaggaa aaggacatgt aattatcatt gactagccca tctcaattgg
     8761 tatagtgatt aaaatcacct agaccaattg agatgtatgt ctgaattagt tgttttcaaa
     8821 gcaaatgaac tagcgattag tcgctatgac ttaacggagc atgaaaccaa gctaatttta
     8881 tgctgtgtgg cactactcaa ccccacgatt gaaaacccta caaggaaaga acggacggta
     8941 tcgttcactt ataaccaata cgctcagatg atgaacatca gtagggaaaa tgcttatggt
     9001 gtattagcta aagcaaccag agagctgatg acgagaactg tggaaatcag gaatcctttg
     9061 gttaaaggct ttgagatttt ccagtggaca aactatgcca agttctcaag cgaaaaatta
     9121 gaattagttt ttagtgaaga gatattgcct tatcttttcc agttaaaaaa attcataaaa
     9181 tataatctgg aacatgttaa gtcttttgaa aacaaatact ctatgaggat ttatgagtgg
     9241 ttattaaaag aactaacaca aaagaaaact cacaaggcaa atatagagat tagccttgat
     9301 gaatttaagt tcatgttaat gcttgaaaat aactaccatg agtttaaaag gcttaaccaa
     9361 tgggttttga aaccaataag taaagattta aacacttaca gcaatatgaa attggtggtt
     9421 gataagcgag gccgcccgac tgatacgttg attttccaag ttgaactaga tagacaaatg
     9481 gatctcgtaa ccgaacttga gaacaaccag ataaaaatga atggtgacaa aataccaaca
     9541 accattacat cagattccta cctacataac ggactaagaa aaacactaca cgatgcttta
     9601 actgcaaaaa ttcagctcac cagttttgag gcaaaatttt tgagtgacat gcaaagtaag
     9661 tatgatctca atggttcgtt ctcatggctc acgcaaaaac aacgaaccac actagagaac
     9721 atactggcta aatacggaag gatctgaggt tcttatggct cttgtatcta tcagtgaagc
     9781 atcaagacta acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa agggaaaact
     9841 gtccatatgc acagatgaaa acggtgtaaa aaagatagat acatcagagc ttttacgagt
     9901 ttttggtgca ttcaaagctg ttcaccatga acagatcgac aatgtaacct cgagattacc
     9961 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
    10021 agcgaggaag cctgcataac gcgaagtaat cttttcggtt ttaaagaaaa agggcagggt
    10081 ggtgacacct tgcccttttt tgccggactg cagcggccgc tactagtagg gagaccagaa
    10141 acaaaaaaag gccccccgtt agggaggcct tcaataattg gctctagtat tattaaactg
    10201 atgcagcgta gttttcgtcg tttgcagcag gccttttgta cagttcatcc ataccatgcg
    10261 tgatgcccgc tgcggttacg aactccagca gaaccatatg atcgcgtttc tcgttcggat
    10321 ctttagacag aacgctttgc gtgctcagat agtgattgtc tggcagcaga acaggaccat
    10381 caccgattgg agtgttttgc tggtagtgat cagccagctg cacgctgcca tcctccacgt
    10441 tgtggcgaat tttaaaattc gctttaatgc catttttttg tttatcggcg g
//
