pSET-dCas9

LOCUS       pSET-dCas9               10158 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of the dCas9 gene in Streptomyces.
ACCESSION   .
VERSION     .
KEYWORDS    pSET-dCas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10158)
  AUTHORS   Zhao Y, Li L, Zheng G, Jiang W, Deng Z, Wang Z, Lu Y
  TITLE     CRISPR/dCas9-Mediated Multiplex Gene Repression in Streptomyces.
  JOURNAL   Biotechnol J. 2018 Jun 3. doi: 10.1002/biot.201800121.
  PUBMED    29862648
REFERENCE   2  (bases 1 to 10158)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10158
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     408..430
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     422..439
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     423..439
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             666..4772
                     /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"
     primer_bind     complement(4951..4967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4951..4967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4964..4986)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4975..4991
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4999..5029)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5044..5065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5182..5199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5353..5941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5433..5452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6145..6945
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             complement(7379..7750)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7783..7892
                     /note="incP origin of transfer"
     CDS             complement(8130..8141)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    8212..8311
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     CDS             8321..10138
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
ORIGIN
        1 atctacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
       61 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
      121 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
      181 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattta tgcggtgtga
      241 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
      301 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      361 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      421 ttgtaaaacg acggccagtg ccaagcttgg gctgcaggtc gactctagac tctagtatgc
      481 atgcgagtgt ccgttcgagt ggcggcttgc gcccgatgct agtcgcggtt gatcggcgat
      541 cgcaggtgca cgcggtcgat cttgacggct ggcgagaggt gcggggagga tctgaccgac
      601 gcggtccaca cgtggcaccg cgatgctgtt gtgggcacaa tcgtgccggt tggtaggatc
      661 cacatatgga caagaagtac tccatcggcc tggcgatcgg caccaactcc gtcggctggg
      721 ccgtcatcac ggacgagtac aaggtcccgt cgaagaagtt caaggtgctg ggcaacacgg
      781 accgccacag catcaagaag aacctcatcg gcgccctcct gttcgacagc ggcgagacgg
      841 ccgaggcgac ccgcctgaag cgcacggcgc ggcggcggta cacgcgccgc aagaaccgca
      901 tctgctacct ccaggagatc ttctccaacg agatggcgaa ggtcgacgac tcgttcttcc
      961 accggctgga ggagtccttc ctggtcgagg aggacaagaa gcacgagcgg caccccatct
     1021 tcggcaacat cgtggacgag gtggcctacc acgagaagta cccgaccatc taccacctcc
     1081 ggaagaagct ggtggactcg accgacaagg ccgacctgcg cctgatctac ctggccctcg
     1141 cccacatgat caagttccgc ggccacttcc tcatcgaggg cgacctgaac ccggacaact
     1201 cggacgtcga caagctgttc atccagctcg tgcagaccta caaccagctc ttcgaggaga
     1261 acccgatcaa cgcctccggc gtggacgcga aggccatcct gtcggcccgc ctctcgaagt
     1321 cccggcggct cgagaacctg atcgcccagc tgccgggcga gaagaagaac ggcctcttcg
     1381 gcaacctgat cgcgctctcc ctgggcctga cgccgaactt caagtccaac ttcgacctcg
     1441 ccgaggacgc caagctccag ctgtccaagg acacgtacga cgacgacctg gacaacctcc
     1501 tggcgcagat cggcgaccag tacgcggacc tgttcctcgc ggcgaagaac ctctccgacg
     1561 cgatcctgct cagcgacatc ctccgcgtca acaccgagat caccaaggcg cccctgtccg
     1621 cgagcatgat caagcgctac gacgagcacc accaggacct caccctgctg aaggccctgg
     1681 tccgccagca gctcccggag aagtacaagg agatcttctt cgaccagtcc aagaacggct
     1741 acgcgggcta catcgacggc ggcgcgtccc aggaggagtt ctacaagttc atcaagccca
     1801 tcctggagaa gatggacggc accgaggagc tgctggtcaa gctgaaccgg gaggacctcc
     1861 tgcgcaagca gcgcacgttc gacaacggct ccatccccca ccagatccac ctgggcgagc
     1921 tgcacgccat cctccggcgc caggaggact tctacccgtt cctcaaggac aaccgcgaga
     1981 agatcgagaa gatcctcacg ttccggatcc cgtactacgt cggccccctc gcccgcggca
     2041 actcccgctt cgcgtggatg acgcgcaagt cggaggagac gatcaccccg tggaacttcg
     2101 aggaggtggt ggacaagggc gcctccgcgc agtcgttcat cgagcggatg accaacttcg
     2161 acaagaacct gccgaacgag aaggtgctgc ccaagcacag cctcctgtac gagtacttca
     2221 cggtctacaa cgagctcacc aaggtgaagt acgtgacgga gggcatgcgg aagccggcgt
     2281 tcctgtccgg cgagcagaag aaggccatcg tggacctgct gttcaagacg aaccggaagg
     2341 tcaccgtcaa gcagctgaag gaggactact tcaagaagat cgagtgcttc gactcggtcg
     2401 agatcagcgg cgtggaggac cgcttcaacg cgtcgctggg cacgtaccac gacctcctga
     2461 agatcatcaa ggacaaggac ttcctcgaca acgaggagaa cgaggacatc ctggaggaca
     2521 tcgtgctgac cctgacgctc ttcgaggacc gggagatgat cgaggagcgc ctcaagacct
     2581 acgcgcacct gttcgacgac aaggtcatga agcagctcaa gcggcggcgg tacaccggct
     2641 ggggccggct ctcccgcaag ctcatcaacg gcatccggga caagcagtcg ggcaagacca
     2701 tcctcgactt cctgaagtcc gacggcttcg ccaaccgcaa cttcatgcag ctgatccacg
     2761 acgacagcct gacgttcaag gaggacatcc agaaggcgca ggtgtcgggc cagggcgact
     2821 ccctgcacga gcacatcgcg aacctggccg gcagcccggc catcaagaag ggcatcctgc
     2881 agacggtcaa ggtggtcgac gagctggtga aggtgatggg ccgccacaag cccgagaaca
     2941 tcgtcatcga gatggcccgg gagaaccaga ccacgcagaa gggccagaag aacagccgcg
     3001 agcgcatgaa gcggatcgag gagggcatca aggagctcgg ctcgcagatc ctcaaggagc
     3061 accccgtcga gaacacccag ctccagaacg agaagctcta cctctactac ctgcagaacg
     3121 gccgggacat gtacgtggac caggagctcg acatcaaccg gctgtcggac tacgacgtgg
     3181 acgccatcgt cccgcagtcg ttcctgaagg acgacagcat cgacaacaag gtcctcaccc
     3241 gctcggacaa gaaccggggc aagtccgaca acgtcccgtc ggaggaggtc gtgaagaaga
     3301 tgaagaacta ctggcggcag ctcctcaacg ccaagctcat cacgcagcgc aagttcgaca
     3361 acctgacgaa ggcggagcgg ggcggcctca gcgagctcga caaggcgggc ttcatcaagc
     3421 ggcagctggt cgagacgcgg cagatcacga agcacgtggc ccagatcctg gactcgcgca
     3481 tgaacaccaa gtacgacgag aacgacaagc tgatccgcga ggtcaaggtc atcaccctca
     3541 agtccaagct ggtgagcgac ttccgcaagg acttccagtt ctacaaggtg cgggagatca
     3601 acaactacca ccacgcccac gacgcgtacc tgaacgcggt ggtgggcacg gcgctgatca
     3661 agaagtaccc gaagctcgag agcgagttcg tctacggcga ctacaaggtg tacgacgtgc
     3721 ggaagatgat cgcgaagagc gagcaggaga tcggcaaggc cacggcgaag tacttcttct
     3781 acagcaacat catgaacttc ttcaagaccg agatcaccct ggcgaacggc gagatccgca
     3841 agcgcccgct catcgagacc aacggcgaga cgggcgagat cgtgtgggac aagggccgcg
     3901 acttcgcgac cgtccggaag gtgctgtcca tgccccaggt caacatcgtc aagaagacgg
     3961 aggtccagac cggcggcttc agcaaggagt ccatcctgcc caagcggaac agcgacaagc
     4021 tgatcgcccg gaagaaggac tgggacccca agaagtacgg cggcttcgac tcccccaccg
     4081 tcgcctactc ggtcctcgtg gtcgccaagg tggagaaggg caagagcaag aagctgaagt
     4141 ccgtgaagga gctgctgggc atcaccatca tggagcggtc gtcgttcgag aagaacccca
     4201 tcgacttcct ggaggccaag ggctacaagg aggtgaagaa ggacctcatc atcaagctcc
     4261 ccaagtactc cctgttcgag ctggagaacg gccggaagcg catgctcgcc agcgcgggcg
     4321 agctgcagaa gggcaacgag ctggcgctgc cgagcaagta cgtcaacttc ctgtacctcg
     4381 cctcgcacta cgagaagctg aagggctcgc cggaggacaa cgagcagaag cagctgttcg
     4441 tggagcagca caagcactac ctcgacgaga tcatcgagca gatctccgag ttctccaagc
     4501 gcgtcatcct ggccgacgcg aacctggaca aggtcctgtc cgcgtacaac aagcaccgcg
     4561 acaagccgat ccgggagcag gccgagaaca tcatccacct gttcacgctg accaacctcg
     4621 gcgcccccgc ggccttcaag tacttcgaca ccacgatcga ccggaagcgg tacacgagca
     4681 cgaaggaggt gctcgacgcc accctgatcc accagagcat cacgggcctg tacgagaccc
     4741 gcatcgacct ctcccagctg ggcggcgact gaatttaaat atctaaagtt ttgtcgtctt
     4801 tccagacgtt agtaaatgaa ttttctgtat gaggttttgc taaacaactt tcaacagttt
     4861 cagcggagtg agaatagaaa ggaacaacta aaggaattgc gaataataat tttttcacgt
     4921 tgaaaatctc gatatcgaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
     4981 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5041 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5101 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5161 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5221 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5281 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5341 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5401 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5521 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5701 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5761 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5821 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5881 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5941 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6001 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tggttcatgt
     6061 gcagctccat cagcaaaagg ggatgataag tttatcacca ccgactattt gcaacagtgc
     6121 cgttgatcgt gctatgatcg actgatgtca tcagcggtgg agtgcaatgt cgtgcaatac
     6181 gaatggcgaa aagccgagct catcggtcag cttctcaacc ttggggttac ccccggcggt
     6241 gtgctgctgg tccacagctc cttccgtagc gtccggcccc tcgaagatgg gccacttgga
     6301 ctgatcgagg ccctgcgtgc tgcgctgggt ccgggaggga cgctcgtcat gccctcgtgg
     6361 tcaggtctgg acgacgagcc gttcgatcct gccacgtcgc ccgttacacc ggaccttgga
     6421 gttgtctctg acacattctg gcgcctgcca aatgtaaagc gcagcgccca tccatttgcc
     6481 tttgcggcag cggggccaca ggcagagcag atcatctctg atccattgcc cctgccacct
     6541 cactcgcctg caagcccggt cgcccgtgtc catgaactcg atgggcaggt acttctcctc
     6601 ggcgtgggac acgatgccaa cacgacgctg catcttgccg agttgatggc aaaggttccc
     6661 tatggggtgc cgagacactg caccattctt caggatggca agttggtacg cgtcgattat
     6721 ctcgagaatg accactgctg tgagcgcttt gccttggcgg acaggtggct caaggagaag
     6781 agccttcaga aggaaggtcc agtcggtcat gcctttgctc ggttgatccg ctcccgcgac
     6841 attgtggcga cagccctggg tcaactgggc cgagatccgt tgatcttcct gcatccgcca
     6901 gaggcgggat gcgaagaatg cgatgccgct cgccagtcga ttggctgagc tcatgagcgg
     6961 agaacgagat gacgttggag gggcaaggtc gcgctgattg ctggggcaac acgtggagcg
     7021 gatcggggat tgtctttctt cagctcgctg atgatatgct gacgctcaat gccgtttggc
     7081 ctccgactaa cgaaaatccc gcatttggac ggctgatccg attggcacgg cggacggcga
     7141 atggcggagc agacgctcgt ccgggggcaa tgagatatga aaaagcctga actcaccgcg
     7201 acgtatcggg ccctggccag ctagctagag tcgacctgca ggtccccggg gatcggtctt
     7261 gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct tgatggagcg
     7321 catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc
     7381 atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct
     7441 cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt
     7501 cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg
     7561 ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg
     7621 ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc
     7681 gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt
     7741 catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag
     7801 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     7861 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     7921 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     7981 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagatc cgtcgacctg
     8041 caggcatgca agctctagcg attccagacg tcccgaaggc gtggcgcggc ttccccgtgc
     8101 cggagcaatc gccctgggtg ggttacacga cgcccctcta tggcccgtac tgacggacac
     8161 accgaagccc cggcggcaac cctcagcgga tgccccgggg cttcacgttt tcccaggtca
     8221 gaagcggttt tcgggagtag tgccccaact ggggtaacct ttgagttctc tcagttgggg
     8281 gcgtagggtc gccgacatga cacaaggggt tgtgaccggg gtggacacgt acgcgggtgc
     8341 ttacgaccgt cagtcgcgcg agcgcgaaaa ttcgagcgca gcaagcccag cgacacagcg
     8401 tagcgccaac gaagacaagg cggccgacct tcagcgcgaa gtcgagcgcg acgggggccg
     8461 gttcaggttc gtcgggcatt tcagcgaagc gccgggcacg tcggcgttcg ggacggcgga
     8521 gcgcccggag ttcgaacgca tcctgaacga atgccgcgcc gggcggctca acatgatcat
     8581 tgtctatgac gtgtcgcgct tctcgcgcct gaaggtcatg gacgcgattc cgattgtctc
     8641 ggaattgctc gccctgggcg tgacgattgt ttccactcag gaaggcgtct tccggcaggg
     8701 aaacgtcatg gacctgattc acctgattat gcggctcgac gcgtcgcaca aagaatcttc
     8761 gctgaagtcg gcgaagattc tcgacacgaa gaaccttcag cgcgaattgg gcgggtacgt
     8821 cggcgggaag gcgccttacg gcttcgagct tgtttcggag acgaaggaga tcacgcgcaa
     8881 cggccgaatg gtcaatgtcg tcatcaacaa gcttgcgcac tcgaccactc cccttaccgg
     8941 acccttcgag ttcgagcccg acgtaatccg gtggtggtgg cgtgagatca agacgcacaa
     9001 acaccttccc ttcaagccgg gcagtcaagc cgccattcac ccgggcagca tcacggggct
     9061 ttgtaagcgc atggacgctg acgccgtgcc gacccggggc gagacgattg ggaagaagac
     9121 cgcttcaagc gcctgggacc cggcaaccgt tatgcgaatc cttcgggacc cgcgtattgc
     9181 gggcttcgcc gctgaggtga tctacaagaa gaagccggac ggcacgccga ccacgaagat
     9241 tgagggttac cgcattcagc gcgacccgat cacgctccgg ccggtcgagc ttgattgcgg
     9301 accgatcatc gagcccgctg agtggtatga gcttcaggcg tggttggacg gcagggggcg
     9361 cggcaagggg ctttcccggg ggcaagccat tctgtccgcc atggacaagc tgtactgcga
     9421 gtgtggcgcc gtcatgactt cgaagcgcgg ggaagaatcg atcaaggact cttaccgctg
     9481 ccgtcgccgg aaggtggtcg acccgtccgc acctgggcag cacgaaggca cgtgcaacgt
     9541 cagcatggcg gcactcgaca agttcgttgc ggaacgcatc ttcaacaaga tcaggcacgc
     9601 cgaaggcgac gaagagacgt tggcgcttct gtgggaagcc gcccgacgct tcggcaagct
     9661 cactgaggcg cctgagaaga gcggcgaacg ggcgaacctt gttgcggagc gcgccgacgc
     9721 cctgaacgcc cttgaagagc tgtacgaaga ccgcgcggca ggcgcgtacg acggacccgt
     9781 tggcaggaag cacttccgga agcaacaggc agcgctgacg ctccggcagc aaggggcgga
     9841 agagcggctt gccgaacttg aagccgccga agccccgaag cttccccttg accaatggtt
     9901 ccccgaagac gccgacgctg acccgaccgg ccctaagtcg tggtgggggc gcgcgtcagt
     9961 agacgacaag cgcgtgttcg tcgggctctt cgtagacaag atcgttgtca cgaagtcgac
    10021 tacgggcagg gggcagggaa cgcccatcga gaagcgcgct tcgatcacgt gggcgaagcc
    10081 gccgaccgac gacgacgaag acgacgccca ggacggcacg gaagacgtag cggcgtagcg
    10141 agacacccgg gaagcctg
//