LOCUS       sequence-257362-        9391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dcas9 generator and reporter circuit (PpspA-2G6 with sfgfp::ASV).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9391)
  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 9391)
  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: pLY152
FEATURES             Location/Qualifiers
     source          1..9391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    364..384
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      385..442
                     /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(466..483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(745..1695)
                     /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      1743..1965
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2457..2551)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2581..3372,3373..3441))
                     /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     3204..3223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3442..3546)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3611..3629
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3633..3676)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3679..3700
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(3764..5350,5351..5353,5354..7840,7841..7843,
                     7844..7870))
                     /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             7877..7888
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        7906..7961
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    7907..7925
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             7977..8603
                     /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(8053..8072)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             8618..8629
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        8835..8884
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             8917..8928
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(8935..9391,1..257)
                     /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 ccgccgataa acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg
       61 atggcagcgt gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg
      121 ttctgctgcc agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg
      181 agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta
      241 tggatgaact gtacaaaagg cctgctgcaa acgacgaaaa ctacgctgca tcagtttaat
      301 aatactagag ccaattattg aaggcctccc taacgggggg cctttttttg tttctggtct
      361 ccctactagt agcggccgct gcagtccggc aaaaaagggc aaggtgtcac caccctgccc
      421 tttttcttta aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc tcactgactc
      481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatctc
      541 gaggttacat tgtcgatctg ttcatggtga acagctttga atgcaccaaa aactcgtaaa
      601 agctctgatg tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc
      661 tttgatatgt aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg
      721 atagatacaa gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag
      781 tgtggttcgt tgtttttgcg tgagccatga gaacgaacca ttgagatcat acttactttg
      841 catgtcactc aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc
      901 gtgtagtgtt tttcttagtc cgttatgtag gtaggaatct gatgtaatgg ttgttggtat
      961 tttgtcacca ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct
     1021 atctagttca acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa
     1081 tttcatattg ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag
     1141 ccttttaaac tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct
     1201 aatctctata tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata
     1261 aatcctcata gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa
     1321 tttttttaac tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc
     1381 gcttgagaac ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt
     1441 cctgatttcc acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc
     1501 attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg
     1561 ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag
     1621 cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac
     1681 aactaattca gacatacatc tcaattggtc taggtgattt taatcactat accaattgag
     1741 atgggctagt caatgataat tacatgtcct tttcctttga gttgtgggta tctgtaaatt
     1801 ctgctagacc tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag
     1861 acctttgtgt gttttttttg tttatattca agtggttata atttatagaa taaagaaaga
     1921 ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt
     1981 tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta
     2041 aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc aaatcgctga atattccttt
     2101 tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc
     2161 tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg
     2221 caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg
     2281 cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt
     2341 ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc
     2401 agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtccctagt gcttggattc
     2461 tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct
     2521 gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgtaaactt ggtctgacag
     2581 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2641 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2701 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2761 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2821 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2881 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2941 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3001 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3061 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3121 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3181 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3241 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3301 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3361 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3421 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3481 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3541 tccgcgcaca tttcccatgg tgccacctga cgtctaagaa accattatta tcatgacatt
     3601 aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt
     3661 cgctaaggat gatttctgga attcgcggcc gcttctagag gggagaccag aaacaaaaaa
     3721 aggggagcgg tttcccgctc cccttcaata attggctcta gtattagtca cctcctagct
     3781 gactcaaatc aatgcgtgtt tcataaagac cagtgatgga ttgatggata agagtggcat
     3841 ctaaaacttc ttttgtagac gtatatcgtt tacgatcaat tgttgtatca aaatatttaa
     3901 aagcagcggg agctccaaga ttcgtcaacg taaataaatg aataatattt tctgcttgtt
     3961 cacgtattgg tttgtctcta tgtttgttat atgcactaag aactttatct aaattggcat
     4021 ctgctaaaat aacacgctta gaaaattcac tgatttgctc aataatctca tctaaataat
     4081 gcttatgctg ctccacaaac aattgttttt gttcgttatc ttctggacta cccttcaact
     4141 tttcataatg actagctaaa tataaaaaat tcacatattt gcttggcaga gccagctcat
     4201 ttcctttttg taattctccg gcactagcca gcatccgttt acgaccgttt tctaactcaa
     4261 aaagactata tttaggtagt ttaatgatta agtctttttt aacttcctta tatcctttag
     4321 cttctaaaaa gtcaatcgga tttttttcaa aggaacttct ttccataatt gtgatcccta
     4381 gtaactcttt aacggatttt aacttcttcg atttcccttt ttccacctta gcaaccacta
     4441 ggactgaata agctaccgtt ggactatcaa aaccaccata tttttttgga tcccagtctt
     4501 ttttacgagc aataagcttg tccgaatttc tttttggtaa aattgactcc ttggagaatc
     4561 cgcctgtctg tacttctgtt ttcttgacaa tattgacttg gggcatggac aatactttgc
     4621 gcactgtggc aaaatctcgc cctttatccc agacaatttc tccagtttcc ccattagttt
     4681 cgattagagg gcgtttgcga atctctccat ttgcaagtgt aatttctgtt ttgaagaagt
     4741 tcatgatatt agagtaaaag aaatattttg cggttgcttt gcctatttct tgctcagact
     4801 tagcaatcat tttacgaaca tcataaactt tataatcacc atagacaaac tccgattcaa
     4861 gttttggata tttcttaatc aaagcagttc caacgacggc atttagatac gcatcatggg
     4921 catgatggta attgttaatc tcacgtactt tatagaattg gaaatctttt cggaagtcag
     4981 aaactaattt agattttaag gtaatcactt taacctctcg aataagttta tcattttcat
     5041 cgtatttagt attcatgcga ctatccaaaa tttgtgccac atgcttagtg atttggcgag
     5101 tttcaaccaa ttggcgtttg ataaaaccag ctttatcaag ttcactcaaa cctccacgtt
     5161 cagctttcgt taaattatca aacttacgtt gagtgattaa cttggcgttt agaagttgtc
     5221 tccaatagtt tttcatcttt ttgactactt cttcacttgg aacgttatcc gatttaccac
     5281 gatttttatc agaacgcgtt aagaccttat tgtctattga atcgtcttta aggaaacttt
     5341 gtggaacaat ggcatcgaca tcataatcac ttaaacgatt aatatctaat tcttggtcca
     5401 catacatgtc tcttccattt tggagataat agagatagag cttttcattt tgcaattgag
     5461 tattttcaac aggatgctct ttaagaatct gacttcctaa ttctttgata ccttcttcga
     5521 ttcgtttcat acgctctcgc gaatttttct ggcccttttg agttgtctga ttttcacgtg
     5581 ccatttcaat aacgatattt tctggcttat gccgccccat tactttgacc aattcatcaa
     5641 caacttttac agtctgtaaa ataccttttt taatagcagg gctaccagct aaatttgcaa
     5701 tatgttcatg taaactatcg ccttgtccag acacttgtgc tttttgaatg tcttctttaa
     5761 atgtcaaact atcatcatgg atcagctgca taaaattgcg attggcaaaa ccatctgatt
     5821 tcaaaaaatc taatattgtt ttgccagatt gcttatccct aataccatta atcaattttc
     5881 gagacaaacg tccccaacca gtataacggc gacgtttaag ctgtttcatc accttatcat
     5941 caaagaggtg agcatatgtt ttaagtcttt cctcaatcat ctccctatct tcaaataagg
     6001 tcaatgttaa aacaatatcc tctaagatat cttcattttc ttcattatcc aaaaaatctt
     6061 tatctttaat aatttttagc aaatcatggt aggtacctaa tgaagcatta aatctatctt
     6121 caactcctga aatttcaaca ctatcaaaac attctatttt tttgaaataa tcttctttta
     6181 attgcttaac ggttactttt cgatttgttt tgaagagtaa atcaacaatg gctttcttct
     6241 gttcacctga aagaaatgct ggttttcgca ttccttcagt aacatatttg acctttgtca
     6301 attcgttata aaccgtaaaa tactcataaa gcaaactatg ttttggtagt actttttcat
     6361 ttggaagatt tttatcaaag tttgtcatgc gttcaataaa tgattgagct gaagcacctt
     6421 tatcgacaac ttcttcaaaa ttccatgggg taattgtttc ttcagacttc cgagtcatcc
     6481 atgcaaaacg actattgcca cgcgccaatg gaccaacata ataaggaata cgaaaagtca
     6541 agattttttc aatcttctca cgattgtctt ttaaaaatgg ataaaagtct tcttgtcttc
     6601 tcaaaatagc atgcagctca cccaagtgaa tttgatgggg aatagagccg ttgtcaaagg
     6661 tccgttgctt gcgcagcaaa tcttcacgat ttagtttcac caataattcc tcagtaccat
     6721 ccattttttc taaaattggt ttgataaatt tataaaattc ttcttggcta gctcccccat
     6781 caatataacc tgcatatccg ttttttgatt gatcaaaaaa gatttcttta tacttttctg
     6841 gaagttgttg tcgaactaaa gcttttaaaa gagtcaagtc ttgatgatgt tcatcgtagc
     6901 gtttaatcat tgaagctgat aggggagcct tagttatttc agtatttact cttaggatat
     6961 ctgaaagtaa aatagcatct gataaattct tagctgccaa aaacaaatca gcatattgat
     7021 ctccaatttg cgccaataaa ttatctaaat catcatcgta agtatctttt gaaagctgta
     7081 atttagcatc ttctgccaaa tcaaaatttg atttaaaatt aggggtcaaa cccaatgaca
     7141 aagcaatgag attcccaaat aagccatttt tcttctcacc ggggagctga gcaatgagat
     7201 tttctaatcg tcttgattta ctcaatcgtg cagaaagaat cgctttagca tctactccac
     7261 ttgcgttaat agggttttct tcaaataatt gattgtaggt ttgtaccaac tggataaata
     7321 gtttgtccac atcactatta tcaggattta aatctccctc aatcaaaaaa tgaccacgaa
     7381 acttaatcat atgcgctaag gccaaataga ttaagcgcaa atccgcttta tcagtagaat
     7441 ctaccaattt ttttcgcaga tgatagatag ttggatattt ctcatgataa gcaacttcat
     7501 ctactatatt tccaaaaata ggatgacgtt catgcttctt gtcttcttcc accaaaaaag
     7561 actcttcaag tcgatgaaag aaactatcat ctactttcgc catctcattt gaaaaaatct
     7621 cctgtagata acaaatacga ttcttccgac gtgtatacct tctacgagct gtccgtttga
     7681 ggcgagtcgc ttccgctgtc tctccactgt caaataaaag agcccctata agattttttt
     7741 tgatactgtg gcggtctgta tttcccagaa ccttgaactt tttagacgga accttatatt
     7801 catcagtgat caccgcccat ccgacgctat ttgtgccgat agctaagcct attgagtatt
     7861 tcttatccat agatcctttc tcctctttag atcttttcaa ttcttttctc tatcactgat
     7921 agggagtggt aaaataactc tatcaacgat agagtgtcaa caaaaattag gaattaatga
     7981 tgtcaagatt agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg
     8041 gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat
     8101 tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt gagatgttag
     8161 ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat tttttacgta
     8221 ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca aaagtacatt
     8281 taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta gcctttttat
     8341 gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg gggcatttta
     8401 ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa agggaaacac
     8461 ctactactga tagtatgccg ccattattac gacaagctat cgaattattt gatcaccaag
     8521 gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta gaaaaacaac
     8581 ttaaatgtga aagtgggtct taatactaga gccaattatt gaaggccgct aacgcggcct
     8641 ttttttgttt ctggtctccc tactagaaaa gctgttgtgg caaaaaacgg aaagctgttg
     8701 tggcaaaaaa cggcatcctg gcatgttgct gttgattctt caatcagatc tttataaatc
     8761 aaaaagataa aaaattggca cgcaaattgt attaacagtt cagcaggaca atcctgaacg
     8821 caaatactag agagctgtca ccggatgtgc tttccggtct gatgagtccg tgaggacgaa
     8881 acagcctcta caaataattt tgtttaacta gagattaaag aggagaaata ccatatgcgt
     8941 aaaggcgaag agctgttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc
     9001 aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg
     9061 acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg
     9121 acgctgactt atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac
     9181 ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat
     9241 gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc
     9301 attgagctga aaggcattga ctttaaagaa gacggcaata tcctgggcca taagctggaa
     9361 tacaatttta acagccacaa tgtttacatc a
//
