LOCUS       sequence-468848-       11706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Streptomyces containing gene of modified Cas9-BD protein
            and sequence of sgRNA cloning template.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11706)
  AUTHORS   Kim DG, Gu B, Cha Y, Ha J, Lee Y, Kim G, Cho BK, Oh MK
  TITLE     Engineered CRISPR-Cas9 for Streptomyces sp. genome editing to 
            improve specialized metabolite production.
  JOURNAL   Nat Commun. 2025 Jan 21;16(1):874. doi: 10.1038/s41467-025-56278-y.
  PUBMED    39833194
REFERENCE   2  (bases 1 to 11706)
  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: "2025-01-21"; volume: "16(1):874. doi"; pages: " 
            10.1038/s41467-025-56278-y"
COMMENT     Sequence Label: pCRISPomyces-2BD
FEATURES             Location/Qualifiers
     source          1..11706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(116..487)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            520..629
                     /note="incP origin of transfer"
     promoter        917..1218
                     /label=rpsL promoter
                     /note="promoter for the Xylanimonas cellulosilytica 
                     ribosomal protein S12 gene"
     CDS             join(1267..1290,1291..1293,1294..5367)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(5429..5445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5453..5469
                     /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(join(5477..5483,5484..5501,5502..5507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5522..5543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5660..5677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5930..6214
                     /label=gapdh promoter
                     /note="glyceraldehyde-3-phosphate dehydrogenase promoter 
                     from Eggerthella lenta"
     CDS             complement(6223..6519)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     6369..6391
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6383..6400
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6384..6400
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6410..6429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6410..6428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6476..6496)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6476..6494)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6515..6531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6539..6555
                     /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(join(6563..6569,6570..6587,6588..6593))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6608..6629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        6672..6747
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      6769..6803
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      complement(6845..7433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6925..6944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7637..8437
                     /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(9376..10818)
                     /codon_start=1
                     /gene="rep"
                     /product="replication initiator protein from the 
                     Streptomyces ghanaensis plasmid pSG5"
                     /label=pSG5 Rep
                     /translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
                     AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
                     IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
                     SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
                     GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
                     RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
                     PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
                     ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
                     APVLQPWGQHPGDCLRVDTAVA"
ORIGIN
        1 atgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg ctcttcttga tggagcgcat
       61 ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa aaaagtcatg
      121 gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc tgcttctctt
      181 cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg cgcgggtcgt
      241 cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg atgcgggcca
      301 gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg ccgacggcca
      361 gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc gctcttcgtt
      421 cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc ttggtttcat
      481 cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag
      541 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
      601 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
      661 ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa
      721 aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagatccgt cgacctgcag
      781 gcatgcgaat tcgttaccgc tgctggtatc acccacggta tggacgaact gtacaaagat
      841 gcatgccagt tctagcataa ccctaatgag tgagctaacc tacattaatt gcgttgcgcc
      901 ttaattaacg aagcttgccc tgcaggcgga agtcaggtag acacgacttc cgctagtcct
      961 tgcaaggtct gctgacgtga ggcggggcgg tcgtttttga ccgccctgcc ttcgtcatgt
     1021 aggctcgctc gctgtgcctg gcgtgtcatc agacgcccag gtcccggtgc cgtgaggccc
     1081 gggccatcga gccggtggta cgtggctgcg gtccccttgt gagggctgcg cgccgtgtgc
     1141 tgtccggcgc gcacagcctt gaatccaccc gcgggggccg gccggtgagc gtgagctcga
     1201 gtagacgacg gagacgtaat ggacgacgac gacgacggcg gcggcggctc cgggggcggg
     1261 ggcagcgaca agaagtacag catcggcctg gccatcggca ccaacagcgt gggctgggcg
     1321 gtcatcaccg acgagtacaa ggtcccctcc aagaagttca aggtcctggg caacaccgac
     1381 cggcactcga tcaagaagaa cctgatcggc gccctgctct tcgacagcgg cgaaaccgcc
     1441 gaggcgaccc gcctgaagcg gaccgcccgt cgccgctaca cccggcgcaa gaaccgcatc
     1501 tgctacctgc aggagatctt ctccaacgag atggccaagg tcgacgactc gttcttccac
     1561 cggctcgagg agagcttcct ggtggaggag gacaagaagc acgagcgcca cccgatcttc
     1621 ggcaacatcg tcgacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctccgc
     1681 aagaagctgg tcgactcgac cgacaaggcg gacctgcggc tcatctacct ggccctcgcg
     1741 cacatgatca agttccgcgg ccacttcctc atcgagggcg acctgaaccc ggacaactcc
     1801 gacgtcgaca agctcttcat ccagctggtg cagacctaca accagctgtt cgaggagaac
     1861 cccatcaacg ccagcggcgt cgacgccaag gcgatcctct ccgcgcgcct gagcaagtcc
     1921 cggcgcctgg agaacctcat cgcccagctg ccgggcgaga agaagaacgg cctcttcggc
     1981 aacctgatcg cgctgtcgct cggcctgacc cccaacttca agagcaactt cgacctggcc
     2041 gaggacgcga agctccagct gtccaaggac acctacgacg acgacctgga caacctgctc
     2101 gcccagatcg gcgaccagta cgcggacctc ttcctggccg cgaagaacct ctcggacgcc
     2161 atcctgctca gcgacatcct gcgggtcaac accgagatca ccaaggcccc gctgtcggcg
     2221 agcatgatca agcggtacga cgagcaccac caggacctga ccctgctcaa ggccctcgtg
     2281 cgccagcagc tgcccgagaa gtacaaggag atcttcttcg accagtccaa gaacggctac
     2341 gccggctaca tcgacggcgg cgcgtcgcag gaggagttct acaagttcat caagccgatc
     2401 ctggagaaga tggacggcac cgaggagctg ctcgtcaagc tgaaccgcga ggacctgctc
     2461 cgcaagcagc ggaccttcga caacggctcc atcccgcacc agatccacct gggcgagctc
     2521 cacgccatcc tccggcgcca ggaggacttc taccccttcc tgaaggacaa ccgcgagaag
     2581 atcgagaaga tcctgacctt ccggatcccg tactacgtcg gccccctggc ccgcggcaac
     2641 tcccggttcg cgtggatgac ccggaagtcg gaggaaacca tcaccccgtg gaacttcgag
     2701 gaggtcgtgg acaagggcgc ctccgcgcag tcgttcatcg agcgcatgac caacttcgac
     2761 aagaacctcc cgaacgagaa ggtcctgccc aagcacagcc tgctctacga gtacttcacc
     2821 gtgtacaacg agctgaccaa ggtcaagtac gtgaccgagg gcatgcggaa gccggccttc
     2881 ctgtccggcg agcagaagaa ggcgatcgtc gacctgctct tcaagaccaa ccgcaaggtc
     2941 accgtgaagc agctgaagga ggactacttc aagaagatcg agtgcttcga ctccgtcgag
     3001 atctcgggcg tggaggaccg cttcaacgcc tccctgggca cctaccacga cctgctcaag
     3061 atcatcaagg acaaggactt cctcgacaac gaggagaacg aggacatcct ggaggacatc
     3121 gtcctcaccc tgaccctctt cgaggaccgc gagatgatcg aggagcggct caagacctac
     3181 gcccacctgt tcgacgacaa ggtgatgaag cagctgaagc ggcgccggta caccggctgg
     3241 ggccgcctct cccggaagct gatcaacggc atccgggaca agcagagcgg caagaccatc
     3301 ctggacttcc tcaagtccga cggcttcgcc aaccgcaact tcatgcagct catccacgac
     3361 gactcgctga ccttcaagga ggacatccag aaggcccagg tgtccggcca gggcgacagc
     3421 ctccacgagc acatcgccaa cctggcgggc tccccggcga tcaagaaggg catcctccag
     3481 accgtcaagg tcgtggacga gctggtcaag gtgatgggcc gccacaagcc cgagaacatc
     3541 gtgatcgaga tggcccggga gaaccagacc acccagaagg gccagaagaa ctcccgcgag
     3601 cggatgaagc gcatcgagga gggcatcaag gagctcggct cgcagatcct gaaggagcac
     3661 ccggtcgaga acacccagct ccagaacgag aagctgtacc tctactacct gcagaacggc
     3721 cgcgacatgt acgtggacca ggagctcgac atcaaccggc tgagcgacta cgacgtcgac
     3781 cacatcgtgc cgcagtcctt cctgaaggac gactcgatcg acaacaaggt cctgacccgc
     3841 tccgacaaga accggggcaa gtccgacaac gtgccctcgg aggaggtcgt gaagaagatg
     3901 aagaactact ggcgccagct gctcaacgcc aagctcatca cccagcgcaa gttcgacaac
     3961 ctgaccaagg ccgagcgggg cggcctgtcg gagctcgaca aggcgggctt catcaagcgc
     4021 cagctcgtcg aaacccggca gatcaccaag cacgtggccc agatcctgga cagccggatg
     4081 aacaccaagt acgacgagaa cgacaagctg atccgcgagg tcaaggtgat caccctcaag
     4141 agcaagctgg tgtccgactt ccgcaaggac ttccagttct acaaggtccg ggagatcaac
     4201 aactaccacc acgcccacga cgcgtacctg aacgccgtcg tgggcaccgc gctgatcaag
     4261 aagtacccga agctggagtc cgagttcgtc tacggcgact acaaggtcta cgacgtgcgc
     4321 aagatgatcg ccaagtcgga gcaggagatc ggcaaggcca ccgcgaagta cttcttctac
     4381 agcaacatca tgaacttctt caagaccgag atcaccctgg ccaacggcga gatccgcaag
     4441 cggcccctga tcgaaaccaa cggcgaaacc ggcgagatcg tctgggacaa gggccgcgac
     4501 ttcgccaccg tccggaaggt gctgtccatg ccgcaggtca acatcgtcaa gaaaaccgag
     4561 gtgcagaccg gcggcttcag caaggagtcc atcctcccca agcgcaactc ggacaagctg
     4621 atcgcccgga agaaggactg ggacccgaag aagtacggcg gcttcgacag ccccaccgtc
     4681 gcctactccg tgctggtcgt ggcgaaggtc gagaagggca agagcaagaa gctgaagtcc
     4741 gtgaaggagc tgctcggcat caccatcatg gagcgctcct cgttcgagaa gaacccgatc
     4801 gacttcctgg aggccaaggg ctacaaggag gtcaagaagg acctcatcat caagctgccc
     4861 aagtactcgc tgttcgagct cgagaacggc cgcaagcgga tgctcgccag cgcgggcgag
     4921 ctgcagaagg gcaacgagct ggccctcccg tccaagtacg tcaacttcct gtacctcgcg
     4981 tcccactacg agaagctgaa gggctcgccc gaggacaacg agcagaagca gctcttcgtg
     5041 gagcagcaca agcactacct ggacgagatc atcgagcaga tctcggagtt cagcaagcgg
     5101 gtcatcctgg ccgacgcgaa cctcgacaag gtgctgtccg cctacaacaa gcaccgcgac
     5161 aagccgatcc gggagcaggc ggagaacatc atccacctgt tcaccctcac caacctgggc
     5221 gcccccgccg cgttcaagta cttcgacacc accatcgacc gcaagcggta caccagcacc
     5281 aaggaggtcc tcgacgcgac cctgatccac cagtccatca ccggcctgta cgaaacccgc
     5341 atcgacctct cccagctcgg cggcgactcg gggggggggg ggagcggcgg ggggggcgac
     5401 gacgacgacg actgaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     5461 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5521 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5581 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5641 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5701 gagcggtatc agctcactca aaggcggtaa caattcagat ctacgcgttc cccgcaaaag
     5761 cggcctttga ctccctgcaa gcctcagcga ccgaatatat cggttatgcg tgggcgatgg
     5821 ttgttgtcat tgtcggcgca actatcggta tcaagctgtt taagaaattc acctcgaaag
     5881 caagctgata aaccgataca attaaaggct ccttttggag cctttttttg ctgctccttc
     5941 ggtcggacgt gcgtctacgg gcaccttacc gcagccgtcg gctgtgcgac acggacggat
     6001 cgggcgaact ggccgatgct gggagaagcg cgctgctgta cggcgcgcac cgggtgcgga
     6061 gcccctcggc gagcggtgtg aaacttctgt gaatggcctg ttcggttgct ttttttatac
     6121 ggctgccaga taaggcttgc agcatctggg cggctaccgc tatgatcggg gcgttcctgc
     6181 aattcttagt gcgagtatct gaaaggggat acgcaagtct tctcagccgc tacagggcgc
     6241 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6301 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     6361 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
     6421 actatagggc gaattgggta ccgggccccc cctcgaggtc ctccagcttt tgttcccttt
     6481 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6541 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6601 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccacc
     6661 ggtgaagaca tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     6721 cttgaaaaag tggcaccgag tcggtgcttt ttactccatc tggatttgtt cagaacgctc
     6781 ggttgccgcc gggcgttttt tatctagagg ccaggaaccg taaaaaggcc gcgttgctgg
     6841 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6901 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6961 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7021 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7081 gctccaagct gggctgtgtg cacgaccccc ccgttcagcc cgaccgctgc gccttatccg
     7141 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7201 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7261 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7321 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7381 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7441 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7501 tggtcatgag attatcaaaa aggatcttca cctagatcct tttggttcat gtgcagctcc
     7561 atcagcaaaa ggggatgata agtttatcac caccgactat ttgcaacagt gccgttgatc
     7621 gtgctatgat cgactgatgt catcagcggt ggagtgcaat gtcgtgcaat acgaatggcg
     7681 aaaagccgag ctcatcggtc agcttctcaa ccttggggtt acccccggcg gtgtgctgct
     7741 ggtccacagc tccttccgta gcgtccggcc cctcgaagat gggccacttg gactgatcga
     7801 ggccctgcgt gctgcgctgg gtccgggagg gacgctcgtc atgccctcgt ggtcaggtct
     7861 ggacgacgag ccgttcgatc ctgccacgtc gcccgttaca ccggaccttg gagttgtctc
     7921 tgacacattc tggcgcctgc caaatgtaaa gcgcagcgcc catccatttg cctttgcggc
     7981 agcggggcca caggcagagc agatcatctc tgatccattg cccctgccac ctcactcgcc
     8041 cgcaagcccg gtcgcccgtg tccatgaact cgatgggcag gtacttctcc tcggcgtggg
     8101 acacgatgcc aacacgacgc tgcatcttgc cgagttgatg gcaaaggttc cctatggggt
     8161 gccgagacac tgcaccattc ttcaggatgg caagttggta cgcgtcgatt atctcgagaa
     8221 tgaccactgc tgtgagcgct ttgccttggc ggacaggtgg ctcaaggaga agagccttca
     8281 gaaggaaggt ccagtcggtc atgcctttgc tcggttgatc cgctcccgcg acattgtggc
     8341 gacagccctg ggtcaactgg gccgagatcc gttgatcttc ctgcatccgc cagaggcggg
     8401 atgcgaagaa tgcgatgccg ctcgccagtc gattggctga gctcatgagc ggagaacgag
     8461 atgacgttgg aggggcaagg tcgcgctgat tgctggggca acacgtggag cggatcgggg
     8521 attgtctttc ttcagctcgc tgatgatatg ctgacgctca atgccgtttg gcctccgact
     8581 aacgaaaatc ccgcatttgg acggctgatc cgattggcac ggcggacggc gaatggcgga
     8641 gcagacgctc gtccgggggc aatgagatat gaaaaagcct gaactcaccg cgacgtatcg
     8701 atgtcgaggt tcctcagggg agccacccca gagaagccct cggagctgag cggagctatt
     8761 tccaaagcca tgccagctag agacagtgca caccgccaag catctgcaaa accctcgcct
     8821 ggagggaaag tgcaatgtac gtacgctggt ttccctccag aaagggattc tgcggcttct
     8881 caactttgga agaagaggcg gaacgaactg ctgtttcagc ctactcatgt gaggaggctg
     8941 gtcctttacc ctgatgcccg agggcatcag ggtaaaggac cagcctcgcc aactaggagc
     9001 gctaccaagg cgaacacaac ccaggacttg gtagaaccct tcgccggaga agttcagact
     9061 catcggcatg agcacccctc gccgcgcact cggcaccggt ccgtcgacca ccaccaggac
     9121 gtcgttgtcg acgtcggccc cgcggctcct gcccgccgaa cgcgtcgtcg tcgacggcct
     9181 ggtgctcatc gacgagcacc cggagccagg tgaaaagcgc cggcggacgc tcggactggg
     9241 cgcgggattc cagcagtaac ccaggtccgc cggccacctc acggcaggca gaccctcggc
     9301 tttcgcgccg ggaccgccat gagaccgcca cccggatgtc cggggtggcg gtctcatggc
     9361 ggtccctcag cggccctacg cgaccgctgt gtcgacgcgg aggcagtctc cggggtgctg
     9421 gccccagggc tggagtaccg gggcgagctt gcccttgcag cggcggcaca cgggcgacgc
     9481 ggcgccgggc gggggagtct tgacgtcgac ggtgaccggc ttcggcttca ggcgcttccg
     9541 aaggctgatc gtcgggcgga tcgcctcctt cttcggctga cgcactcggt cgagcgaagc
     9601 ggccagctcc tccgtgcggg cctcgttggc cttccggcgc gcctcgcgga aagcagcttc
     9661 ctcctcggac atgacctcga acctcatctg gtcagcgtca aggtcgcccg gcgcggccgg
     9721 cgcttccggg gcgggcgggt ccaggacgtc cttgccccac accaggcccc aggactcgac
     9781 gagccgccgg acgcccggta ggccgtacgt ctcggcgacc ttgatgaggt cgaggcgacg
     9841 tccggcgacg cgggcgatgt atcggtacca gatgtaggcc gggatgaccg cgatggcgac
     9901 caggccctcg gtgtcgtcgg tgatctcctc ctcggtgcgg acgtcctgct ggatgccgag
     9961 ttccttgatc agccggttca ggttctgcga ccggtagtgc ttgcggacct ggaacacgcc
    10021 gaactcgcgc tcgcggtact tctcgacgaa cgggccgggc cgacgaagcc gctgcagctc
    10081 ggcggccgcc gcgtcgccca ggtcgagcgg tcccatgcgg tcgtcgccgc gaccggcctt
    10141 gaagttctgt ccggccagct ccaggccgat cttggcgacg ccgcccttgg tcttgtcgcc
    10201 gtccttgtag aggtagcggg cctgcttgcc cgcatcgccg tcagcggcgt ccgcgccgtt
    10261 gagtgggcgc acgtcggtgc cgtggccctt gccctcacag gagcaaccgg gccggtcgca
    10321 cgtctcgctg acggtgtagc cgcccgcgga ttcgaccccg gcggcccagg ctccggcgag
    10381 tgcgtcgcgg aacgcggcct gggcgtccgg gccgagcacc tcgcgggtga cccagagcgt
    10441 gtgccagtgc aggtgccagc cggagcccca gccgaaggtg tcctcgaagg cccgctcgta
    10501 gccgatgatc ccgaagtcgt cgcgcatcgt gcgccagcgg cggccggacg agccgtacgc
    10561 gcccttccag ccgtcgtgca agaccgcgac caggccgtgc cgcattccct tgcggacggt
    10621 gccgaacgcc atgcgctcga agtggcgcaa cgtgttcgtg ccaaggtgca gcccgtaccc
    10681 ggcgtccgcg agaccgtcgg cggcgagctg cacgttcgag ccccgtacgg ccaggatgcg
    10741 gctcatgcac cacgggcagg tgtggacgtt gttgcagcgg cacgtgttgc cccacgtcgc
    10801 ctcgcccggc ttccacatca gctcggccgt cccggcagtg agccgggtcc cgcagccctt
    10861 gaacgcctcg ttcagcgaca ccgtctggtg ccggtcccgc cgggcgaacc gctcgtcgcg
    10921 cgggtcctcc cgcccggctg tcgcggcacc ctcgtttggg gtagaacccg ttccagttac
    10981 agcgctctga cctgcagtgg acggagattt tcccttacta ctaaagcccg cgtccggatt
    11041 acccgctgta gtcgtgcttg ctacgctgcg tgactggtcc gcaatgagac gctttgcgcg
    11101 ctttcggcag gcgtccgagc agtagatttt ggggcgcttc ccggggatgt ggacgatcgg
    11161 ggtgccgcag tggcacttcg gtccggcggg gcgcggtggt gtcgacgcgc tgttctctcg
    11221 tacgctcgtc acagagcaaa cgtcctcact cggcatgctg cgccggttcg ggggcggcga
    11281 gcccgggagg ccaatcccgg gctcgtgcca tttctgggtc ctgttgatcc tggcattggt
    11341 gtggccgttc attgcccctg ctcgctcctg acgcgccgat agacgtccga tacgcccggt
    11401 gctggtggga tttgataggt cggaagaagc cccgcccggg cctgggcggg gcttcctgtg
    11461 cgtcaggacc tcctcgtcgt gagcctcttc ggcctatgga cggagtgacc tcgtgatccg
    11521 ttacagccgc gcgcgctcgc gtagagcggt ctcatcagtt ccacgaacgg tcctcttcgc
    11581 agatcagggc gttggggcgg agtctcacca aggactacgt ctgctggcga tttccgttac
    11641 accccgggcg gtggccggcg cacacgcgcg cccgcgttgg gcagtgcaga aagtgcagaa
    11701 acctag
//
