LOCUS       sequence-257326-       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-LEA4B4 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: pLY56
FEATURES             Location/Qualifiers
     source          1..10491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             463..474
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(507..556)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             complement(778..789)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(1788..1822)
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             complement(1862..1873)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(1888..2514)
                     /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     2419..2438
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        2530..2585
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    2566..2584
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2603..2614
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2621..2647,2648..2650,2651..5137,5138..5140,
                     5141..6727)
                     /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    6791..6812
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      6815..6858
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(6862..6880)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6945..7049
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7050..7118,7119..7910)
                     /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(7268..7287)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7940..8034
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      8526..8748
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             8796..9746
                     /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     10008..10025
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(10049..10106)
                     /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    10107..10127
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(join(10234..10491,1..456))
                     /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 gatgtaaaca ttgtggctgt taaaattgta ttccagctta tggcccagga tattgccgtc
       61 ttctttaaag tcaatgcctt tcagctcaat gcggtttacc agggtatcgc cttcaaattt
      121 cacttccgca cgcgttttgt acgtgccgtc atccttaaag gaaatcgtgc gttcctgcac
      181 atagccttcc ggcatggcgg acttgaagaa gtcatgctgc ttcatatggt ccggataacg
      241 agcaaagcac tgaacaccat aagtcagcgt cgttaccaga gtcggccaag gtaccggcag
      301 tttaccagta gtacagatga acttcagcgt cagtttacca ttagttgcgt caccttcacc
      361 ctcgccacgc acggaaaact tatgaccgtt gacatcacca tccagttcca ccagaatagg
      421 gacgacacca gtgaacagct cttcgccttt acgcatatgg tatttctcct ctttaatctc
      481 tagttaaaca aaattatttg tagaggctgt ttcgtcctca cggactcatc agaccggaaa
      541 gcacatccgg tgacagctct ctagtatttg cgttcaggat tgtcctgctg aactgttaat
      601 acaatttgcg tgccaatttt ttatcttttt gatttataaa gatctgattg aagaatcaac
      661 agcaacatgc caggatgagt tagcgaatta cactaaccgt tatttccgaa cgatcaccgg
      721 ggaacgaagc gagataactt ctagtaggga gaccagaaac aaaaaaaggc cgcgttagcg
      781 gccttcaata attggctcta gtattaattt gctgctttcc attgagcctg tttctctgcg
      841 cgacgttcgc ggcggcgtgt gcgtgcattc atttgcaaac tgagttgcag caactctttt
      901 tcctgctgca tctgaaactc acgtaaatcc agcggcagtg ttggaagcga ggtggtttct
      961 gaaacggcga tagcgtcttc aggcggacgc cgtttaaagg gatcaataat gatgtcatca
     1021 agcggataat cgctggtgcc gtggcgatac actgaacgtt ccaccacgtt tttcaattca
     1081 cgaatatttc ccggccaacg ataattcagc aatgtttctc tggcgcgctc cgtaaacccc
     1141 gggaacagag gcagcttgat ttcccgacac atctggatgg caaagtattc tgccatcaac
     1201 attatgtcgc tttcgcgctc gcgcagtggt ggcagttgta caacatcaaa agccagtcgg
     1261 tcgagcaggt cagcgcgaaa agtgccttca ttgaccatcg ccgggagatc ggcattcgtc
     1321 gcgcatacca accgcacatt cacctgcaat ggttggctgc cgccaacgcg ctccagttca
     1381 ccgtactcaa tcacgcgcaa taatttctcc tgcaccatca tcggtgccgt agcgagttca
     1441 tcaagaaata gcgtaccgcc gtcggcacgt tcaaatctcc ctggatgacg tttttgcgca
     1501 ccggtaaacg cccccgcttc gtgaccaaac agttcggaat ccagcagatt ttcatttaac
     1561 gccgcgcagt taagggaaat aaacggccct tgccaacggg aggagagata atgcaggcgg
     1621 ctggcaatca gctctttacc ggtgccgcgt tcgccgatga tgagcaccgg tttgtccagc
     1681 ggtgcgagat gcgaaacctg ttccagcact tcgagaaagc tgttcgcctc accaagtaaa
     1741 ttatctttgt attctgccat ctagtacttt cctgtgtgac tctagtagct agcactatac
     1801 ctaggactga gctagccgta aactctagta gggagaccag aaacaaaaaa aggccgcgtt
     1861 agcggccttc aataattggc tctagtatta agacccactt tcacatttaa gttgtttttc
     1921 taatccgcat atgatcaatt caaggccgaa taagaaggct ggctctgcac cttggtgatc
     1981 aaataattcg atagcttgtc gtaataatgg cggcatacta tcagtagtag gtgtttccct
     2041 ttcttcttta gcgacttgat gctcttgatc ttccaatacg caacctaaag taaaatgccc
     2101 cacagcgctg agtgcatata atgcattctc tagtgaaaaa ccttgttggc ataaaaaggc
     2161 taattgattt tcgagagttt catactgttt ttctgtaggc cgtgtaccta aatgtacttt
     2221 tgctccatcg cgatgactta gtaaagcaca tctaaaactt ttagcgttat tacgtaaaaa
     2281 atcttgccag ctttcccctt ctaaagggca aaagtgagta tggtgcctat ctaacatctc
     2341 aatggctaag gcgtcgagca aagcccgctt attttttaca tgccaataca atgtaggctg
     2401 ctctacacct agcttctggg cgagtttacg ggttgttaaa ccttcgattc cgacctcatt
     2461 aagcagctct aatgcgctgt taatcacttt acttttatct aatcttgaca tcattaattc
     2521 ctaatttttg ttgacactct atcgttgata gagttatttt accactccct atcagtgata
     2581 gagaaaagaa ttgaaaagat ctaaagagga gaaaggatct atggataaga aatactcaat
     2641 aggcttagct atcggcacaa atagcgtcgg atgggcggtg atcactgatg aatataaggt
     2701 tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc cacagtatca aaaaaaatct
     2761 tataggggct cttttatttg acagtggaga gacagcggaa gcgactcgcc tcaaacggac
     2821 agctcgtaga aggtatacac gtcggaagaa tcgtatttgt tatctacagg agattttttc
     2881 aaatgagatg gcgaaagtag atgatagttt ctttcatcga cttgaagagt cttttttggt
     2941 ggaagaagac aagaagcatg aacgtcatcc tatttttgga aatatagtag atgaagttgc
     3001 ttatcatgag aaatatccaa ctatctatca tctgcgaaaa aaattggtag attctactga
     3061 taaagcggat ttgcgcttaa tctatttggc cttagcgcat atgattaagt ttcgtggtca
     3121 ttttttgatt gagggagatt taaatcctga taatagtgat gtggacaaac tatttatcca
     3181 gttggtacaa acctacaatc aattatttga agaaaaccct attaacgcaa gtggagtaga
     3241 tgctaaagcg attctttctg cacgattgag taaatcaaga cgattagaaa atctcattgc
     3301 tcagctcccc ggtgagaaga aaaatggctt atttgggaat ctcattgctt tgtcattggg
     3361 tttgacccct aattttaaat caaattttga tttggcagaa gatgctaaat tacagctttc
     3421 aaaagatact tacgatgatg atttagataa tttattggcg caaattggag atcaatatgc
     3481 tgatttgttt ttggcagcta agaatttatc agatgctatt ttactttcag atatcctaag
     3541 agtaaatact gaaataacta aggctcccct atcagcttca atgattaaac gctacgatga
     3601 acatcatcaa gacttgactc ttttaaaagc tttagttcga caacaacttc cagaaaagta
     3661 taaagaaatc ttttttgatc aatcaaaaaa cggatatgca ggttatattg atgggggagc
     3721 tagccaagaa gaattttata aatttatcaa accaatttta gaaaaaatgg atggtactga
     3781 ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga cctttgacaa
     3841 cggctctatt ccccatcaaa ttcacttggg tgagctgcat gctattttga gaagacaaga
     3901 agacttttat ccatttttaa aagacaatcg tgagaagatt gaaaaaatct tgacttttcg
     3961 tattccttat tatgttggtc cattggcgcg tggcaatagt cgttttgcat ggatgactcg
     4021 gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa gttgtcgata aaggtgcttc
     4081 agctcaatca tttattgaac gcatgacaaa ctttgataaa aatcttccaa atgaaaaagt
     4141 actaccaaaa catagtttgc tttatgagta ttttacggtt tataacgaat tgacaaaggt
     4201 caaatatgtt actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac agaagaaagc
     4261 cattgttgat ttactcttca aaacaaatcg aaaagtaacc gttaagcaat taaaagaaga
     4321 ttatttcaaa aaaatagaat gttttgatag tgttgaaatt tcaggagttg aagatagatt
     4381 taatgcttca ttaggtacct accatgattt gctaaaaatt attaaagata aagatttttt
     4441 ggataatgaa gaaaatgaag atatcttaga ggatattgtt ttaacattga ccttatttga
     4501 agatagggag atgattgagg aaagacttaa aacatatgct cacctctttg atgataaggt
     4561 gatgaaacag cttaaacgtc gccgttatac tggttgggga cgtttgtctc gaaaattgat
     4621 taatggtatt agggataagc aatctggcaa aacaatatta gattttttga aatcagatgg
     4681 ttttgccaat cgcaatttta tgcagctgat ccatgatgat agtttgacat ttaaagaaga
     4741 cattcaaaaa gcacaagtgt ctggacaagg cgatagttta catgaacata ttgcaaattt
     4801 agctggtagc cctgctatta aaaaaggtat tttacagact gtaaaagttg ttgatgaatt
     4861 ggtcaaagta atggggcggc ataagccaga aaatatcgtt attgaaatgg cacgtgaaaa
     4921 tcagacaact caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa tcgaagaagg
     4981 tatcaaagaa ttaggaagtc agattcttaa agagcatcct gttgaaaata ctcaattgca
     5041 aaatgaaaag ctctatctct attatctcca aaatggaaga gacatgtatg tggaccaaga
     5101 attagatatt aatcgtttaa gtgattatga tgtcgatgcc attgttccac aaagtttcct
     5161 taaagacgat tcaatagaca ataaggtctt aacgcgttct gataaaaatc gtggtaaatc
     5221 ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa aactattgga gacaacttct
     5281 aaacgccaag ttaatcactc aacgtaagtt tgataattta acgaaagctg aacgtggagg
     5341 tttgagtgaa cttgataaag ctggttttat caaacgccaa ttggttgaaa ctcgccaaat
     5401 cactaagcat gtggcacaaa ttttggatag tcgcatgaat actaaatacg atgaaaatga
     5461 taaacttatt cgagaggtta aagtgattac cttaaaatct aaattagttt ctgacttccg
     5521 aaaagatttc caattctata aagtacgtga gattaacaat taccatcatg cccatgatgc
     5581 gtatctaaat gccgtcgttg gaactgcttt gattaagaaa tatccaaaac ttgaatcgga
     5641 gtttgtctat ggtgattata aagtttatga tgttcgtaaa atgattgcta agtctgagca
     5701 agaaataggc aaagcaaccg caaaatattt cttttactct aatatcatga acttcttcaa
     5761 aacagaaatt acacttgcaa atggagagat tcgcaaacgc cctctaatcg aaactaatgg
     5821 ggaaactgga gaaattgtct gggataaagg gcgagatttt gccacagtgc gcaaagtatt
     5881 gtccatgccc caagtcaata ttgtcaagaa aacagaagta cagacaggcg gattctccaa
     5941 ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa aagactggga
     6001 tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct tattcagtcc tagtggttgc
     6061 taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt aaagagttac tagggatcac
     6121 aattatggaa agaagttcct ttgaaaaaaa tccgattgac tttttagaag ctaaaggata
     6181 taaggaagtt aaaaaagact taatcattaa actacctaaa tatagtcttt ttgagttaga
     6241 aaacggtcgt aaacggatgc tggctagtgc cggagaatta caaaaaggaa atgagctggc
     6301 tctgccaagc aaatatgtga attttttata tttagctagt cattatgaaa agttgaaggg
     6361 tagtccagaa gataacgaac aaaaacaatt gtttgtggag cagcataagc attatttaga
     6421 tgagattatt gagcaaatca gtgaattttc taagcgtgtt attttagcag atgccaattt
     6481 agataaagtt cttagtgcat ataacaaaca tagagacaaa ccaatacgtg aacaagcaga
     6541 aaatattatt catttattta cgttgacgaa tcttggagct cccgctgctt ttaaatattt
     6601 tgatacaaca attgatcgta aacgatatac gtctacaaaa gaagttttag atgccactct
     6661 tatccatcaa tccatcactg gtctttatga aacacgcatt gatttgagtc agctaggagg
     6721 tgactaatac tagagccaat tattgaaggg gagcgggaaa ccgctcccct ttttttgttt
     6781 ctggtctccc ctctagaagc ggccgcgaat tccagaaatc atccttagcg aaagctaagg
     6841 atttttttta tctgaaattc tgcctcgtga tacgcctatt tttataggtt aatgtcatga
     6901 taataatggt ttcttagacg tcaggtggca ccatgggaaa tgtgcgcgga acccctattt
     6961 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     7021 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     7081 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     7141 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     7201 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     7261 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     7321 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     7381 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     7441 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     7501 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     7561 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     7621 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7681 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7741 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7801 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7861 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7921 aagtttacga gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat
     7981 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc
     8041 actagggaca gtaagacggg taagcctgtt gatgataccg ctgccttact gggtgcatta
     8101 gccagtctga atgacctgtc acgggataat ccgaagtggt cagactggaa aatcagaggg
     8161 caggaactgc tgaacagcaa aaagtcagat agcaccacat agcagacccg ccataaaacg
     8221 ccctgagaag cccgtgacgg gcttttcttg tattatgggt agtttccttg catgaatcca
     8281 taaaaggcgc ctgtagtgcc atttaccccc attcactgcc agagccgtga gcgcagcgaa
     8341 ctgaatgtca cgaaaaagac agcgactcag gtgcctgatg gtcggagaca aaaggaatat
     8401 tcagcgattt gcccgagctt gcgagggtgc tacttaagcc tttagggttt taaggtctgt
     8461 tttgtagagg agcaaacagc gtttgcgaca tccttttgta atactgcgga actgactaaa
     8521 gtagtgagtt atacacaggg ctgggatcta ttctttttat ctttttttat tctttcttta
     8581 ttctataaat tataaccact tgaatataaa caaaaaaaac acacaaaggt ctagcggaat
     8641 ttacagaggg tctagcagaa tttacaagtt ttccagcaaa ggtctagcag aatttacaga
     8701 tacccacaac tcaaaggaaa aggacatgta attatcattg actagcccat ctcaattggt
     8761 atagtgatta aaatcaccta gaccaattga gatgtatgtc tgaattagtt gttttcaaag
     8821 caaatgaact agcgattagt cgctatgact taacggagca tgaaaccaag ctaattttat
     8881 gctgtgtggc actactcaac cccacgattg aaaaccctac aaggaaagaa cggacggtat
     8941 cgttcactta taaccaatac gctcagatga tgaacatcag tagggaaaat gcttatggtg
     9001 tattagctaa agcaaccaga gagctgatga cgagaactgt ggaaatcagg aatcctttgg
     9061 ttaaaggctt tgagattttc cagtggacaa actatgccaa gttctcaagc gaaaaattag
     9121 aattagtttt tagtgaagag atattgcctt atcttttcca gttaaaaaaa ttcataaaat
     9181 ataatctgga acatgttaag tcttttgaaa acaaatactc tatgaggatt tatgagtggt
     9241 tattaaaaga actaacacaa aagaaaactc acaaggcaaa tatagagatt agccttgatg
     9301 aatttaagtt catgttaatg cttgaaaata actaccatga gtttaaaagg cttaaccaat
     9361 gggttttgaa accaataagt aaagatttaa acacttacag caatatgaaa ttggtggttg
     9421 ataagcgagg ccgcccgact gatacgttga ttttccaagt tgaactagat agacaaatgg
     9481 atctcgtaac cgaacttgag aacaaccaga taaaaatgaa tggtgacaaa ataccaacaa
     9541 ccattacatc agattcctac ctacataacg gactaagaaa aacactacac gatgctttaa
     9601 ctgcaaaaat tcagctcacc agttttgagg caaaattttt gagtgacatg caaagtaagt
     9661 atgatctcaa tggttcgttc tcatggctca cgcaaaaaca acgaaccaca ctagagaaca
     9721 tactggctaa atacggaagg atctgaggtt cttatggctc ttgtatctat cagtgaagca
     9781 tcaagactaa caaacaaaag tagaacaact gttcaccgtt acatatcaaa gggaaaactg
     9841 tccatatgca cagatgaaaa cggtgtaaaa aagatagata catcagagct tttacgagtt
     9901 tttggtgcat tcaaagctgt tcaccatgaa cagatcgaca atgtaacctc gagattaccg
     9961 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
    10021 gcgaggaagc ctgcataacg cgaagtaatc ttttcggttt taaagaaaaa gggcagggtg
    10081 gtgacacctt gccctttttt gccggactgc agcggccgct actagtaggg agaccagaaa
    10141 caaaaaaagg ccccccgtta gggaggcctt caataattgg ctctagtatt attaaactga
    10201 tgcagcgtag ttttcgtcgt ttgcagcagg ccttttgtac agttcatcca taccatgcgt
    10261 gatgcccgct gcggttacga actccagcag aaccatatga tcgcgtttct cgttcggatc
    10321 tttagacaga acgctttgcg tgctcagata gtgattgtct ggcagcagaa caggaccatc
    10381 accgattgga gtgttttgct ggtagtgatc agccagctgc acgctgccat cctccacgtt
    10441 gtggcgaatt ttaaaattcg ctttaatgcc atttttttgt ttatcggcgg t
//
