LOCUS       sequence-418472-       13123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one cumate-based inducible CRISPRi plasmid for use in 
            Streptomyces.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13123)
  AUTHORS   Bai C, van Wezel GP
  TITLE     CUBIC: A Versatile Cumate-Based Inducible CRISPRi System in 
            Streptomyces.
  JOURNAL   ACS Synth Biol. 2023 Oct 6. doi: 10.1021/acssynbio.3c00464.
  PUBMED    37801665
REFERENCE   2  (bases 1 to 13123)
  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: "ACS Synth 
            Biol. 2023 Oct 6. doi: 10.1021/acssynbio.3c00464."
COMMENT     Sequence Label: pCB-1
FEATURES             Location/Qualifiers
     source          1..13123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             168..779
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    908..935
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     misc_feature    951..972
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    1073..1094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1109..1114,1115..1132,1133..1139)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1147..1163
                     /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)."
     RBS             1181..1192
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1199..1876
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1921..1992
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2008..2035
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2042..2062
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     misc_RNA        2070..2145
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    2290..2317
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     CDS             join(2342..2368,2369..2371,2372..4858,4859..4861,
                     4862..6448)
                     /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"
     terminator      6600..6648
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     terminator      6791..6885
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      7377..7599
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             7647..8597
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(10324..10695)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            10728..10837
                     /note="incP origin of transfer"
     CDS             complement(11835..11846)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             13026..13037
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 agtcagtatc cagtcgtgta gcagaatcaa caataaggcc tccctaacgg ggggcctttt
       61 ttattgataa caaaagtatg ttcacattcg aaccgtctct gctttgacaa tggcttgaat
      121 tggggtgtaa agtcgtggcc acctaacgag gagatcggtt catgatcatg tcccccaagc
      181 ggcggaccca ggcggagcgg gcgatggaaa cccagggcaa gctgatcgcg gcggccctcg
      241 gcgtcctgcg cgagaagggc tacgccggct tccggatcgc ggacgtcccg ggcgccgcgg
      301 gcgtgtcgcg cggcgcccag agccaccact tccccaccaa gctcgagctg ctcctggcga
      361 ccttcgagtg gctgtacgag cagatcaccg agcgctcccg ggcccgcctc gcgaagctga
      421 agccggagga cgacgtcatc cagcagatgc tcgacgacgc cgcggagttc ttcctggacg
      481 acgacttctc catctcgctc gacctgatcg tcgccgcgga ccgggacccc gccctgcgcg
      541 agggcatcca gcggaccgtg gagcggaacc gcttcgtcgt ggaggacatg tggctcggcg
      601 tcctggtgag ccggggcctg tcccgcgacg acgccgagga catcctctgg ctgatcttca
      661 actccgtccg cggcctcgcc gtgcggtcgc tgtggcagaa ggacaaggag cggttcgagc
      721 gcgtgcggaa cagcaccctg gagatcgccc gggagcggta cgccaagttc aagcggtgat
      781 agactaatct tttcgaaaaa aggcctccca aatcgggggg ccttttttta tagcaacaaa
      841 agtatgttca cattcgaacc gtctctgctt tgacatcgtg tggcgcttgg gtgtaaagtc
      901 gtggccaaac aaacagacaa tctggtctgt ttgtattatg ccaagagacc gaattcgcgg
      961 ccgcttctag agcaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1021 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     1081 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     1141 gtggaattgt gagcggataa caatttcaca catactagag aaagaggaga aatactagat
     1201 ggcttcctcc gaagacgtta tcaaagagtt catgcgtttc aaagttcgta tggaaggttc
     1261 cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac
     1321 ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt gggacatcct
     1381 gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg acatcccgga
     1441 ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga acttcgaaga
     1501 cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt tcatctacaa
     1561 agttaaactg cgtggtacca acttcccgtc cgacggtccg gttatgcaga aaaaaaccat
     1621 gggttgggaa gcttccaccg aacgtatgta cccggaagac ggtgctctga aaggtgaaat
     1681 caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta aaaccaccta
     1741 catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca aactggacat
     1801 cacctcccac aacgaagact acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca
     1861 ctccaccggt gcttaataac gctgatagtg ctagtgtaga tcgctactag agccaggcat
     1921 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     1981 gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
     2041 atactagtag cggccgctgc agggtctctg ttttagagct agaaatagca agttaaaata
     2101 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tttagtgcca
     2161 tcctcggtac caaattccag aaaagaggcc tcccgaaagg ggggcctttt ttcgttttgg
     2221 tccgtatgtt cacattcgaa ccgtctctgc tttgacatcg tgtggcgctt gggtgtaaag
     2281 tcgtggccaa acaaacagac aatctggtct gtttgtatta ttctaagtaa ggagtgtcca
     2341 tatggacaag aagtactcca tcggcctcgc gatcggcacc aactccgtgg gctgggcggt
     2401 catcaccgac gagtacaagg tcccctccaa gaagttcaag gtcctgggca acaccgaccg
     2461 gcactcgatc aagaagaacc tgatcggcgc cctgctgttc gacagcggcg agacggccga
     2521 ggcgacccgc ctgaagcgga ccgcgcgtcg ccgctacacc cggcgcaaga accgcatctg
     2581 ctacctgcag gaaatcttct ccaacgagat ggccaaggtg gacgactcgt tcttccaccg
     2641 cctggaggag agcttcctgg tggaggagga caagaagcac gagcgccacc cgatcttcgg
     2701 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctccgcaa
     2761 gaagctggtg gactcgaccg acaaggcgga cctgcggctc atctacctgg ccctcgcgca
     2821 catgatcaag ttccgcggcc acttcctcat cgagggcgac ctgaacccgg acaactccga
     2881 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggagaaccc
     2941 catcaacgcc agcggcgtgg acgccaaggc gatcctctcc gcgcgcctga gcaagtcccg
     3001 gcgcctggag aacctcatcg cccagctgcc gggcgagaag aagaacggcc tcttcggcaa
     3061 cctgatcgcg ctgtcgctcg gcctgacccc caacttcaag agcaacttcg acctggccga
     3121 ggacgcgaag ctccagctgt ccaaggacac ctacgacgac gacctggaca acctgctcgc
     3181 ccagatcggc gaccagtacg cggacctctt cctggccgcg aagaacctct cggacgccat
     3241 cctgctcagc gacatcctgc gggtcaacac cgagatcacc aaggccccgc tgtcggcgag
     3301 catgatcaag cggtacgacg agcaccacca ggacctgacc ctgctcaagg ccctcgtgcg
     3361 ccagcagctg cccgagaagt acaaggaaat cttcttcgac cagtccaaga acggctacgc
     3421 cggctacatc gacggcggcg cgtcgcagga ggagttctac aagttcatca agccgatcct
     3481 ggagaagatg gacggcaccg aggagctgct cgtcaagctg aaccgcgagg acctgctccg
     3541 caagcagcgg accttcgaca acggctccat cccgcaccag atccacctgg gcgagctcca
     3601 cgccatcctc cggcgccagg aggacttcta ccccttcctg aaggacaacc gcgagaagat
     3661 cgagaagatc ctgaccttcc gcatcccgta ctacgtcggc cccctggccc gcggcaactc
     3721 ccggttcgcg tggatgaccc ggaagtcgga ggagacgatc accccgtgga acttcgagga
     3781 ggtcgtggac aagggcgcgt ccgcgcagtc gttcatcgag cgcatgacca acttcgacaa
     3841 gaacctcccg aacgagaagg tcctgcccaa gcactccctg ctctacgagt acttcaccgt
     3901 gtacaacgag ctgaccaagg tcaagtacgt gaccgagggc atgcggaagc cggccttcct
     3961 gtcgggcgag cagaagaagg cgatcgtgga cctgctgttc aagaccaacc gcaaggtcac
     4021 cgtgaagcag ctgaaggagg actacttcaa gaagatcgag tgcttcgact ccgtcgagat
     4081 cagcggcgtg gaggaccgct tcaacgcctc cctgggcacc taccacgacc tgctcaagat
     4141 catcaaggac aaggacttcc tcgacaacga ggagaacgag gacatcctgg aggacatcgt
     4201 cctcaccctg accctcttcg aggaccgcga gatgatcgag gagcggctca agacctacgc
     4261 ccacctgttc gacgacaagg tgatgaagca gctgaagcgt cgccgctaca ccggctgggg
     4321 ccgcctctcc cggaagctga tcaacggcat ccgggacaag cagagcggca agaccatcct
     4381 ggacttcctc aagtccgacg gcttcgccaa ccgcaacttc atgcagctca tccacgacga
     4441 cagcctgacc ttcaaggagg acatccagaa ggcccaggtg tcgggccagg gcgacagcct
     4501 ccacgagcac atcgccaacc tggcgggctc cccggcgatc aagaagggca tcctccagac
     4561 cgtcaaggtc gtggacgagc tggtcaaggt gatgggccgc cacaagcccg agaacatcgt
     4621 gatcgagatg gcccgggaga accagaccac ccagaagggc cagaagaact cgcgcgagcg
     4681 gatgaagcgg atcgaggagg gcatcaagga gctcggcagc cagatcctga aggagcaccc
     4741 ggtcgagaac acccagctgc agaacgagaa gctgtacctc tactacctgc agaacggccg
     4801 cgacatgtac gtggaccagg agctcgacat caaccggctg tccgactacg acgtggacgc
     4861 gatcgtgccg cagtccttcc tgaaggacga ctcgatcgac aacaaggtcc tgacccgctc
     4921 ggacaagaac cggggcaagt ccgacaacgt gccctcggag gaggtcgtga agaagatgaa
     4981 gaactactgg cgccagctgc tcaacgccaa gctcatcacc cagcgcaagt tcgacaacct
     5041 gaccaaggcc gagcggggcg gcctgagcga gctcgacaag gcgggcttca tcaagcgcca
     5101 gctggtcgag acgcggcaga tcaccaagca cgtggcccag atcctggact cccggatgaa
     5161 caccaagtac gacgagaacg acaagctgat ccgcgaggtc aaggtgatca ccctcaagag
     5221 caagctggtg tccgacttcc gcaaggactt ccagttctac aaggtccggg agatcaacaa
     5281 ctaccaccac gcccacgacg cgtacctgaa cgccgtcgtg ggcaccgcgc tgatcaagaa
     5341 gtacccgaag ctggagtccg agttcgtcta cggcgactac aaggtctacg acgtgcgcaa
     5401 gatgatcgcc aagagcgagc aggagatcgg caaggccacc gcgaagtact tcttctactc
     5461 caacatcatg aacttcttca agaccgagat caccctggcc aacggcgaga tccgcaagcg
     5521 gcccctgatc gagacgaacg gcgagacggg cgagatcgtc tgggacaagg gccgcgactt
     5581 cgccaccgtc cggaaggtgc tgtcgatgcc gcaggtcaac atcgtgaaga agaccgaggt
     5641 gcagaccggc ggcttcagca aggagtccat cctccccaag cgcaacagcg acaagctgat
     5701 cgcccggaag aaggactggg acccgaagaa gtacggcggc ttcgacagcc ccaccgtcgc
     5761 ctactccgtg ctggtcgtgg cgaaggtcga gaagggcaag agcaagaagc tgaagtccgt
     5821 gaaggagctg ctcggcatca ccatcatgga gcgctcctcg ttcgagaaga acccgatcga
     5881 cttcctggag gccaagggct acaaggaggt caagaaggac ctcatcatca agctgcccaa
     5941 gtacagcctg ttcgagctgg agaacggccg caagcggatg ctcgcctccg cgggcgagct
     6001 gcagaagggc aacgagctgg ccctcccgtc gaagtacgtc aacttcctgt acctcgcgtc
     6061 ccactacgag aagctgaagg gctcgcccga ggacaacgag cagaagcagc tgttcgtgga
     6121 gcagcacaag cactacctgg acgagatcat cgagcagatc agcgagttca gcaagcgcgt
     6181 catcctggcc gacgcgaacc tcgacaaggt gctgtccgcc tacaacaagc accgcgacaa
     6241 gccgatccgg gagcaggcgg agaacatcat ccacctgttc accctcacca acctgggcgc
     6301 ccccgccgcg ttcaagtact tcgacaccac catcgaccgc aagcggtaca cctccaccaa
     6361 ggaggtcctc gacgcgaccc tgatccacca gagcatcacc ggcctgtacg agacgcgcat
     6421 cgacctgtcc cagctcggcg gcgactgata ggagcgatcc cgcaaaagcg gcctttgact
     6481 ccctgcaagc ctcagcgacc gaatatatcg gttatgcgtg ggcgatggtt gttgtcattg
     6541 tcggcgcaac tatcggtatc aagctgttta agaaattcac ctcgaaagca agctgataaa
     6601 ccgatacaat taaaggctcc ttttggagcc tttttttttg gagattttca acgtgaaaaa
     6661 attattattc gcaattcctt tagttgttcc tttctattct cactccgctg aaactgttga
     6721 aagttgttta gcaaaacctc atacagaaaa ttcagctaga gctgtcagac caagtttacg
     6781 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     6841 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag cactagggac
     6901 agtaagacgg gtaagcctgt tgatgatacc gctgccttac tgggtgcatt agccagtctg
     6961 aatgacctgt cacgggataa tccgaagtgg tcagactgga aaatcagagg gcaggaactg
     7021 ctgaacagca aaaagtcaga tagcaccaca tagcagaccc gccataaaac gccctgagaa
     7081 gcccgtgacg ggcttttctt gtattatggg tagtttcctt gcatgaatcc ataaaaggcg
     7141 cctgtagtgc catttacccc cattcactgc cagagccgtg agcgcagcga actgaatgtc
     7201 acgaaaaaga cagcgactca ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt
     7261 tgcccgagct tgcgagggtg ctacttaagc ctttagggtt ttaaggtctg ttttgtagag
     7321 gagcaaacag cgtttgcgac atccttttgt aatactgcgg aactgactaa agtagtgagt
     7381 tatacacagg gctgggatct attcttttta tcttttttta ttctttcttt attctataaa
     7441 ttataaccac ttgaatataa acaaaaaaaa cacacaaagg tctagcggaa tttacagagg
     7501 gtctagcaga atttacaagt tttccagcaa aggtctagca gaatttacag atacccacaa
     7561 ctcaaaggaa aaggacatgt aattatcatt gactagccca tctcaattgg tatagtgatt
     7621 aaaatcacct agaccaattg agatgtatgt ctgaattagt tgttttcaaa gcaaatgaac
     7681 tagcgattag tcgctatgac ttaacggagc atgaaaccaa gctaatttta tgctgtgtgg
     7741 cactactcaa ccccacgatt gaaaacccta caaggaaaga acggacggta tcgttcactt
     7801 ataaccaata cgctcagatg atgaacatca gtagggaaaa tgcttatggt gtattagcta
     7861 aagcaaccag agagctgatg acgagaactg tggaaatcag gaatcctttg gttaaaggct
     7921 ttgagatttt ccagtggaca aactatgcca agttctcaag cgaaaaatta gaattagttt
     7981 ttagtgaaga gatattgcct tatcttttcc agttaaaaaa attcataaaa tataatctgg
     8041 aacatgttaa gtcttttgaa aacaaatact ctatgaggat ttatgagtgg ttattaaaag
     8101 aactaacaca aaagaaaact cacaaggcaa atatagagat tagccttgat gaatttaagt
     8161 tcatgttaat gcttgaaaat aactaccatg agtttaaaag gcttaaccaa tgggttttga
     8221 aaccaataag taaagattta aacacttaca gcaatatgaa attggtggtt gataagcgag
     8281 gccgcccgac tgatacgttg attttccaag ttgaactaga tagacaaatg gatctcgtaa
     8341 ccgaacttga gaacaaccag ataaaaatga atggtgacaa aataccaaca accattacat
     8401 cagattccta cctacataac ggactaagaa aaacactaca cgatgcttta actgcaaaaa
     8461 ttcagctcac cagttttgag gcaaaatttt tgagtgacat gcaaagtaag tatgatctca
     8521 atggttcgtt ctcatggctc acgcaaaaac aacgaaccac actagagaac atactggcta
     8581 aatacggaag gatctgaggt tcttatggct cttgtatcta tcagtgaagc atcaagacta
     8641 acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa agggaaaact gtccatatgc
     8701 acagatgaaa acggtgtaaa aaagatagat acatcagagc ttttacgagt ttttggtgca
     8761 ttcaaagctg ttcaccatga acagatcgac aatgtaacta ctagagggtt catgtgcagc
     8821 tccatcagca aaaggggatg ataagtttat caccaccgac tatttgcaac agtgccgttg
     8881 atcgtgctat gatcgactgg atcggcgggg cctgggacgg cgcctcgccg tcgcagaacc
     8941 aggcggtggc gtacaccgtc gcctcggtcg gcccgtagag attggcgatc ccgaccgcag
     9001 caccaccgag aacgtccccg acgtggccga ccagcccgtc atcgtcaacg cctgatccgc
     9061 ggtgcggaca ggccgtgtcg cgaccggccg tgcggaatta agccggcccg taccctgtga
     9121 atagaggtcc gctgtgacac aagaatccct gttacttctc gaccgtattg attcggatga
     9181 ttcctacgcg agcctgcgga acgaccagga gttctgggag ccgctggccc gccgagccct
     9241 ggaggagctc gggctgccgg tgccgccggt gctgcgggtg cccggcgaga gcaccaaccc
     9301 cgtactggtc ggcgagcccg gcccggtgat caagctgttc ggcgagcact ggtgcggtcc
     9361 ggagagcctc gcgtcggagt cggaggcgta cgcggtcctg gcggacgccc cggtgccggt
     9421 gccccgcctc ctcggccgcg gcgagctgcg gcccggcacc ggagcctggc cgtggcccta
     9481 cctggtgatg agccggatga ccggcaccac ctggcggtcc gcgatggacg gcacgaccga
     9541 ccggaacgcg ctgctcgccc tggcccgcga actcggccgg gtgctcggcc ggctgcacag
     9601 ggtgccgctg accgggaaca ccgtgctcac cccccattcc gaggtcttcc cggaactgct
     9661 gcgggaacgc cgcgcggcga ccgtcgagga ccaccgcggg tggggctacc tctcgccccg
     9721 gctgctggac cgcctggagg actggctgcc ggacgtggac acgctgctgg ccggccgcga
     9781 accccggttc gtccacggcg acctgcacgg gaccaacatc ttcgtggacc tggccgcgac
     9841 cgaggtcacc gggatcgtcg acttcaccga cgtctatgcg ggagactccc gctacagcct
     9901 ggtgcaactg catctcaacg ccttccgggg cgaccgcgag atcctggccg cgctgctcga
     9961 cggggcgcag tggaagcgga ccgaggactt cgcccacgaa ctgctcgcct tcaccttcct
    10021 gcacgacttc gaggtgttcg aggagacgcc gctggatctc tccggcttca ccgatccgga
    10081 ggaactggcg cagttcctct gggggccgcc ggacaccgcc cccggcgcct gacgccccgg
    10141 gcccgcgccg aagcccgctg ctgcgagcta gctagagtcg acctgggtcc ccggggatcg
    10201 gtcttgcctt gctcgtcggt gatgtacttc accagctccg cgaagtcgct cttcttgatg
    10261 gagcgcatgg ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa
    10321 aagtcatggc tctgccctcg ggcggaccac gcccatcatg accttgccaa gctcgtcctg
    10381 cttctcttcg atcttcgcca gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg
    10441 cgggtcgtcg gtgagccaga gtttcagcag gccgcccagg cggcccaggt cgccattgat
    10501 gcgggccagc tcgcggacgt gctcatagtc cacgacgccc gtgattttgt agccctggcc
    10561 gacggccagc aggtaggccg acaggctcat gccggccgcc gccgcctttt cctcaatcgc
    10621 tcttcgttcg tctggaaggc agtacacctt gataggtggg ctgcccttcc tggttggctt
    10681 ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg
    10741 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
    10801 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
    10861 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
    10921 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa agatccgtcg
    10981 acctgtcacg ccgccgctgt gaacccgttc agcgtctccg gctccgtctc ttcctcgacc
    11041 ttcggcttcg cccagtggat acgcacacgg tcggcgatcg ggggaacccg tccgggccgc
    11101 tgaacgccct tcggaagctt gatcacctcg atccggtcca cgaagagccg gacgaacgcc
    11161 cgacggtctt caaggggcgc tagcgcccac cacgaacccg gtcccgtcgg gtcgctgccc
    11221 cggtcgccca gccattcgcc gatcggcagc acgggggagt cggcggcgtc gagctgacga
    11281 agccgttctt cggccccttc catgcggagc gtcagcgcgg cttcctcttc gagaaacgcg
    11341 cgccgtccca tggcggaccg gtagccgccg ttccgcttgt cttcgtaaag ctcttcgagc
    11401 gccttcacgg cgtccgcgcg ctgtgccatc agttcggagc gctgaccctt caactccggg
    11461 cgttcgtgcg tcgctcccca gcgccgtgcg gcttcgtaca tgagcgctgc cgtgtcgcct
    11521 tcgaggtcgt cagggtcggc agggtcgaag gccgtgatgc gcgcgaagat cgcgccgacg
    11581 atgtacgggt caaggttgtg catcgtgatc gagcatgacg acccgtcatg aacccggcgg
    11641 gggcacttgc acgcgtacga cgacttgtga gcttgatcac cttcgcggac gttgcccgcc
    11701 atggtcgacc cgttgctgta acccgtctcc gggtcgagct ggccggagcc gtagcagtaa
    11761 aggacgtcca ttgccgacag gagcgattgc ccccggtact gacccttccc gcgtcctcga
    11821 ccctgaagcc actcctgaag ttcccaccat tccgccgggg gaatgtacgg ctcgaagccg
    11881 ggcagggtca gcggctccat ggtgaccggg tcgcgcctga tcttgtaatg gctgaagccg
    11941 ccccgcgaac cgtcggcgcg caccttgtat gcgatgtcag cttgataccc ggcaatgcgc
    12001 gggtcgctga gtacgcgctt caaaacgccg gggtcccaat cggaaccggc gcgcttctta
    12061 ccgaccagcg tgccgcgcgt aggcaccttg tcgcggtaca gccgctcaca aagcccgttc
    12121 agcgacccag ggtgaaacga cccggctccg ccacccttga agggcgtatc gcggtgcgtc
    12181 ttgatctcct gccacgccca gcggattacc gccccttcgc tgccgtgtgc gccttcccag
    12241 gtgtgcgcgc tgggcaccag tcggcgaatg gcaaccagct ttccgccgtc ttccgggttc
    12301 ggaaccattt cctcgaccgt gtcgaatccg tacggcgtcg accccgtgtg tccgcccagc
    12361 cgcttcgcca attccttagc gttcgacacg gcgacgctct tgttcttcga ctcatcatgc
    12421 gaagcctgaa ggcgcatgat caggtgaata aggtccatca tttcgccggg gcggaatgtg
    12481 ccttcgttca cgctgacaat ggtcacgccc agccggagca attccgtgac gaccggaata
    12541 atgtccagcg gctcttcgcg gctgaggcgc gaaatgtaat gaacaatgat catgttcatt
    12601 tcccggttcc ggcacatgtc caaaatccgg ttgaactccg gccggtcgac gcccgtgaat
    12661 gccgacgtgc cgggcgcttc gctgaagtga cccagccact tcacctcgac gccgtcgcgc
    12721 gcgtactcct tcgccagcgc ctcagccttc ccccggttcg cggcgcgctg ggtggccggt
    12781 gaagcggtcg aactgttctc ccgttccgcc gactgtcggt cgtagccgcc tgccagaatg
    12841 accatgcctt caaccttcac gctcatgccg ggcaggctat cccgcccaac tgggtaagag
    12901 caaagactaa cactgttggg ctggaactac tcccgaaaac cgcttctgac ctgggaaaac
    12961 gtgaagcccc ggggcatccg ctgagggttg ccgccggggc ttcggtgtgt ccgtcagtac
    13021 gggccataga ggggcgtcgt gtaacccacc cagggcgatt gctccggcac ggggaagccg
    13081 cgccacgcct tcgggacgtc tggaatcgct agagcttgca tgc
//
