pCRISPomyces-SaCas9

LOCUS      pCRISPomyces-SaCas9     10216 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Contains codon-optimized SaCas9 and gRNA for streptomyces.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10216)
  AUTHORS   Yeo WL, Heng E, Tan LL, Lim YW, Lim YH, Hoon S, Zhao H, Zhang MM, 
            Wong FT
  TITLE     Characterization of Cas proteins for CRISPR-Cas editing in 
            streptomycetes.
  JOURNAL   Biotechnol Bioeng. 2019 May 15. doi: 10.1002/bit.27021.
  PUBMED    31090220
REFERENCE   2  (bases 1 to 10216)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            Bioeng. 2019 May 15. doi: 10.1002/bit.27021."
COMMENT     Sequence Label: pCRISPomyces-SaCas9
FEATURES             Location/Qualifiers
     source          1..10216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..302
                     /label=rpsL promoter
                     /note="promoter for the Xylanimonas cellulosilytica 
                     ribosomal protein S12 gene"
     CDS             303..3464
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNE
                     GRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEE
                     FSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE
                     VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFG
                     WKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKF
                     QIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKE
                     IIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKA
                     INLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI
                     KVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENA
                     KYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK
                     QEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRF
                     SVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKER
                     NKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEI
                     FITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKD
                     NDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKK
                     DNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN
                     LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLL
                     NRIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQ
                     IIKKG"
     promoter        3664..3948
                     /label=gapdh promoter
                     /note="glyceraldehyde-3-phosphate dehydrogenase promoter 
                     from Eggerthella lenta"
     primer_bind     4103..4125
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4117..4134
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4118..4134
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4144..4163
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4144..4162
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4210..4230)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4210..4228)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4249..4265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4249..4265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4262..4284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4273..4289
                     /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(4297..4303,4304..4321,4322..4327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4342..4363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        4406..4481
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     terminator      4498..4532
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      complement(4746..5334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4826..4845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5538..6338
                     /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(7277..8719)
                     /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"
     oriT            9886..9995
                     /note="incP origin of transfer"
ORIGIN
        1 gccctgcagg cggaagtcag gtagacacga cttccgctag tccttgcaag gtctgctgac
       61 gtgaggcggg gcggtcgttt ttgaccgccc tgccttcgtc atgtaggctc gctcgctgtg
      121 cctggcgtgt catcagacgc ccaggtcccg gtgccgtgag gcccgggcca tcgagccggt
      181 ggtacgtggc tgcggtcccc ttgtgagggc tgcgcgccgt gtgctgtccg gcgcgcacag
      241 ccttgaatcc acccgcgggg gccggccggt ctccgtgagc tcgagtagac gacggagacg
      301 taatgaagcg caactacatc ctggggctgg acatcggcat cacctccgtc ggctacggca
      361 tcatcgacta cgagacgcgc gacgtcatcg acgccggcgt ccgcctgttc aaggaagcca
      421 acgtcgagaa caacgagggc cgccgctcca agcgcggcgc ccgccgcctg aagcgccgcc
      481 gccgccaccg catccagcgc gtcaagaagc tgctgttcga ctacaacctg ctgaccgacc
      541 actccgagct gtccggcatc aacccgtacg aggcccgcgt caagggcctg tcccagaagc
      601 tgtccgagga agagttctcc gccgccctgc tgcacctggc caagcgccgc ggcgtccaca
      661 acgtcaacga ggtggaagag gacaccggca acgagctgtc caccaaggag cagatcagcc
      721 gcaactccaa ggccctggag gagaagtacg tcgccgagct gcaactggag cggctgaaga
      781 aggacggcga ggtccgcggc tccatcaacc gcttcaagac ctccgactac gtcaaggaag
      841 ccaagcagct gctgaaggtc cagaaggcct accaccagct ggaccagtcc ttcatcgaca
      901 cctacatcga cctgctggag acgcgtcgca cctactacga gggtccgggc gagggctccc
      961 cgttcggctg gaaggacatc aaggagtggt acgagatgct gatgggccac tgcacctact
     1021 tcccggagga gctgcgctcc gtcaagtacg cctacaacgc cgacctgtac aacgccctga
     1081 acgacctgaa caacctggtc atcacccgcg acgagaacga gaagctggag tactacgaga
     1141 agttccagat catcgagaac gtgttcaagc agaagaagaa gccgaccctg aagcagatcg
     1201 ccaaggagat cctggtcaac gaagaggaca tcaagggcta ccgcgtcacc tccaccggca
     1261 agccggagtt caccaacctg aaggtctacc acgacatcaa ggacatcacc gcccgcaagg
     1321 agatcatcga gaacgccgag ctgctggacc agatcgccaa gatcctgacg atctaccagt
     1381 cctccgagga catccaggaa gagctgacca acctgaactc cgagctgacc caggaagaga
     1441 tcgagcagat cagcaacctg aagggctaca ccggcaccca caacctgtcc ctgaaggcca
     1501 tcaacctgat cctggacgag ctgtggcaca ccaacgacaa ccagatcgcc atcttcaacc
     1561 gcctgaagct ggtcccgaag aaggtggacc tgtcccagca gaaggagatc ccgaccaccc
     1621 tggtggacga cttcatcctg tccccggtcg tcaagcgctc cttcatccag tccatcaagg
     1681 tcatcaacgc catcatcaag aagtacggcc tgccgaacga catcatcatc gagctggccc
     1741 gcgagaagaa ctccaaggac gcccagaaga tgatcaacga gatgcagaag cgcaaccgcc
     1801 agaccaacga gcgcatcgag gagatcatcc gcaccaccgg caaggagaac gccaagtacc
     1861 tgatcgagaa gatcaagctg cacgacatgc aggaaggcaa gtgcctgtac tccctggagg
     1921 ccatcccgct ggaggacctg ctgaacaacc cgttcaacta cgaggtggac cacatcatcc
     1981 cgcgctccgt ctccttcgac aactccttca acaacaaggt cctggtcaag caggaagaga
     2041 actccaagaa gggcaaccgc accccgttcc agtacctgtc ctcctccgac tccaagatca
     2101 gctacgagac gttcaagaag cacatcctga acctggccaa gggcaagggc cgcatctcca
     2161 agaccaagaa ggagtacctg ctggaggagc gcgacatcaa ccgcttctcc gtccagaagg
     2221 acttcatcaa ccgcaacctg gtggacaccc ggtacgccac ccgcggcctg atgaacctgc
     2281 tgcgctccta cttccgcgtc aacaacctgg acgtcaaggt caagtccatc aacggcggct
     2341 tcacctcctt cctgcgtcgc aagtggaagt tcaagaagga gcgcaacaag ggctacaagc
     2401 accacgccga ggacgccctg atcatcgcca acgccgactt catcttcaag gagtggaaga
     2461 agctggacaa ggccaagaag gtcatggaga accagatgtt cgaggagaag caggccgagt
     2521 ccatgccgga gatcgagacg gagcaggagt acaaggaaat cttcatcacc ccgcaccaga
     2581 tcaagcacat caaggacttc aaggactaca agtactccca ccgcgtggac aagaagccga
     2641 accgcgagct gatcaacgac accctgtact ccacccgcaa ggacgacaag ggcaacaccc
     2701 tgatcgtcaa caacctgaac ggcctgtacg acaaggacaa cgacaagctg aagaagctga
     2761 tcaacaagtc cccggagaag ctgctgatgt accaccacga cccgcagacc taccagaagc
     2821 tgaagctgat catggagcag tacggcgacg agaagaaccc gctgtacaag tactacgagg
     2881 agacgggcaa ctacctgacg aagtactcca agaaggacaa cggcccggtc atcaagaaga
     2941 tcaagtacta cggcaacaag ctgaacgcgc acctggacat caccgacgac tacccgaact
     3001 cccgcaacaa ggtcgtcaag ctgtccctga agccgtaccg cttcgacgtc tacctggaca
     3061 acggcgtcta caagttcgtc accgtcaaga acctggacgt catcaagaag gagaactact
     3121 acgaggtcaa ctccaagtgc tacgaggaag ccaagaagct gaagaagatc agcaaccagg
     3181 ccgagttcat cgcctccttc tacaacaacg acctgatcaa gatcaacggc gagctgtacc
     3241 gcgtcatcgg cgtcaacaac gacctgctga accgcatcga ggtcaacatg atcgacatca
     3301 cctaccgcga gtacctggag aacatgaacg acaagcgccc gccgcgcatc atcaagacga
     3361 tcgcctccaa gacccagtcc atcaagaagt actccaccga catcctgggc aacctgtacg
     3421 aggtcaagtc caagaagcac ccgcagatca tcaagaaggg ctgagaattc agatctacgc
     3481 gttccccgca aaagcggcct ttgactccct gcaagcctca gcgaccgaat atatcggtta
     3541 tgcgtgggcg atggttgttg tcattgtcgg cgcaactatc ggtatcaagc tgtttaagaa
     3601 attcacctcg aaagcaagct gataaaccga tacaattaaa ggctcctttt ggagcctttt
     3661 tttgctgctc cttcggtcgg acgtgcgtct acgggcacct taccgcagcc gtcggctgtg
     3721 cgacacggac ggatcgggcg aactggccga tgctgggaga agcgcgctgc tgtacggcgc
     3781 gcaccgggtg cggagcccct cggcgagcgg tgtgaaactt ctgtgaatgg cctgttcggt
     3841 tgcttttttt atacggctgc cagataaggc ttgcagcatc tgggcggcta ccgctatgat
     3901 cggggcgttc ctgcaattct tagtgcgagt atctgaaagg ggatacgcaa gtcttctcag
     3961 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggggatcg
     4021 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     4081 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     4141 gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga ggtcctccag
     4201 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     4261 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4321 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4381 gcccgctttc caccggtgaa gacatgtttt agtactctgg aaacagaatc tactaaaaca
     4441 aggcaaaatg ccgtgtttat ctcgtcaact tgttggcgag aactccatct ggatttgttc
     4501 agaacgctcg gttgccgccg ggcgtttttt atctagagct agcatgcata tgaagctttg
     4561 ttcacattcg aacggtctct gctttgacaa catgctgtgc ggtgttgtaa agtcgtggcc
     4621 aggagaatac gacagcgtgc aggactgggg gagttatgac tagtgtgaaa agtcagtatc
     4681 cagtcgtgta gttcttatta ccttctagag gccaggaacc gtaaaaaggc cgcgttgctg
     4741 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4801 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4861 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4921 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4981 cgctccaagc tgggctgtgt gcacgacccc cccgttcagc ccgaccgctg cgccttatcc
     5041 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5101 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5161 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5221 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5281 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5341 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5401 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttggttca tgtgcagctc
     5461 catcagcaaa aggggatgat aagtttatca ccaccgacta tttgcaacag tgccgttgat
     5521 cgtgctatga tcgactgatg tcatcagcgg tggagtgcaa tgtcgtgcaa tacgaatggc
     5581 gaaaagccga gctcatcggt cagcttctca accttggggt tacccccggc ggtgtgctgc
     5641 tggtccacag ctccttccgt agcgtccggc ccctcgaaga tgggccactt ggactgatcg
     5701 aggccctgcg tgctgcgctg ggtccgggag ggacgctcgt catgccctcg tggtcaggtc
     5761 tggacgacga gccgttcgat cctgccacgt cgcccgttac accggacctt ggagttgtct
     5821 ctgacacatt ctggcgcctg ccaaatgtaa agcgcagcgc ccatccattt gcctttgcgg
     5881 cagcggggcc acaggcagag cagatcatct ctgatccatt gcccctgcca cctcactcgc
     5941 ctgcaagccc ggtcgcccgt gtccatgaac tcgatgggca ggtacttctc ctcggcgtgg
     6001 gacacgatgc caacacgacg ctgcatcttg ccgagttgat ggcaaaggtt ccctatgggg
     6061 tgccgagaca ctgcaccatt cttcaggatg gcaagttggt acgcgtcgat tatctcgaga
     6121 atgaccactg ctgtgagcgc tttgccttgg cggacaggtg gctcaaggag aagagccttc
     6181 agaaggaagg tccagtcggt catgcctttg ctcggttgat ccgctcccgc gacattgtgg
     6241 cgacagccct gggtcaactg ggccgagatc cgttgatctt cctgcatccg ccagaggcgg
     6301 gatgcgaaga atgcgatgcc gctcgccagt cgattggctg agctcatgag cggagaacga
     6361 gatgacgttg gaggggcaag gtcgcgctga ttgctggggc aacacgtgga gcggatcggg
     6421 gattgtcttt cttcagctcg ctgatgatat gctgacgctc aatgccgttt ggcctccgac
     6481 taacgaaaat cccgcatttg gacggctgat ccgattggca cggcggacgg cgaatggcgg
     6541 agcagacgct cgtccggggg caatgagata tgaaaaagcc tgaactcacc gcgacgtatc
     6601 gatgtcgagg ttcctcaggg gagccacccc agagaagccc tcggagctga gcggagctat
     6661 ttccaaagcc atgccagcta gagacagtgc acaccgccaa gcatctgcaa aaccctcgcc
     6721 tggagggaaa gtgcaatgta cgtacgctgg tttccctcca gaaagggatt ctgcggcttc
     6781 tcaactttgg aagaagaggc ggaacgaact gctgtttcag cctactcatg tgaggaggct
     6841 ggtcctttac cctgatgccc gagggcatca gggtaaagga ccagcctcgc caactaggag
     6901 cgctaccaag gcgaacacaa cccaggactt ggtagaaccc ttcgccggag aagttcagac
     6961 tcatcggcat gagcacccct cgccgcgcac tcggcaccgg tccgtcgacc accaccagga
     7021 cgtcgttgtc gacgtcggcc ccgcggctcc tgcccgccga acgcgtcgtc gtcgacggcc
     7081 tggtgctcat cgacgagcac ccggagccag gtgaaaagcg ccggcggacg ctcggactgg
     7141 gcgcgggatt ccagcagtaa cccaggtccg ccggccacct cacggcaggc agaccctcgg
     7201 ctttcgcgcc gggaccgcca tgagaccgcc acccggatgt ccggggtggc ggtctcatgg
     7261 cggtccctca gcggccctac gcgaccgctg tgtcgacgcg gaggcagtct ccggggtgct
     7321 ggccccaggg ctggagtacc ggggcgagct tgcccttgca gcggcggcac acgggcgacg
     7381 cggcgccggg cgggggagtc ttgacgtcga cggtgaccgg cttcggcttc aggcgcttcc
     7441 gaaggctgat cgtcgggcgg atcgcctcct tcttcggctg acgcactcgg tcgagcgaag
     7501 cggccagctc ctccgtgcgg gcctcgttgg ccttccggcg cgcctcgcgg aaagcagctt
     7561 cctcctcgga catgacctcg aacctcatct ggtcagcgtc aaggtcgccc ggcgcggccg
     7621 gcgcttccgg ggcgggcggg tccaggacgt ccttgcccca caccaggccc caggactcga
     7681 cgagccgccg gacgcccggt aggccgtacg tctcggcgac cttgatgagg tcgaggcgac
     7741 gtccggcgac gcgggcgatg tatcggtacc agatgtaggc cgggatgacc gcgatggcga
     7801 ccaggccctc ggtgtcgtcg gtgatctcct cctcggtgcg gacgtcctgc tggatgccga
     7861 gttccttgat cagccggttc aggttctgcg accggtagtg cttgcggacc tggaacacgc
     7921 cgaactcgcg ctcgcggtac ttctcgacga acgggccggg ccgacgaagc cgctgcagct
     7981 cggcggccgc cgcgtcgccc aggtcgagcg gtcccatgcg gtcgtcgccg cgaccggcct
     8041 tgaagttctg tccggccagc tccaggccga tcttggcgac gccgcccttg gtcttgtcgc
     8101 cgtccttgta gaggtagcgg gcctgcttgc ccgcatcgcc gtcagcggcg tccgcgccgt
     8161 tgagtgggcg cacgtcggtg ccgtggccct tgccctcaca ggagcaaccg ggccggtcgc
     8221 acgtctcgct gacggtgtag ccgcccgcgg attcgacccc ggcggcccag gctccggcga
     8281 gtgcgtcgcg gaacgcggcc tgggcgtccg ggccgagcac ctcgcgggtg acccagagcg
     8341 tgtgccagtg caggtgccag ccggagcccc agccgaaggt gtcctcgaag gcccgctcgt
     8401 agccgatgat cccgaagtcg tcgcgcatcg tgcgccagcg gcggccggac gagccgtacg
     8461 cgcccttcca gccgtcgtgc aagaccgcga ccaggccgtg ccgcattccc ttgcggacgg
     8521 tgccgaacgc catgcgctcg aagtggcgca acgtgttcgt gccaaggtgc agcccgtacc
     8581 cggcgtccgc gagaccgtcg gcggcgagct gcacgttcga gccccgtacg gccaggatgc
     8641 ggctcatgca ccacgggcag gtgtggacgt tgttgcagcg gcacgtgttg ccccacgtcg
     8701 cctcgcccgg cttccacatc agctcggccg tcccggcagt gagccgggtc ccgcagccct
     8761 tgaacgcctc gttcagcgac accgtctggt gccggtcccg ccgggcgaac cgctcgtcgc
     8821 gcgggtcctc ccgcccggct gtcgcggcac cctcgtttgg ggtagaaccc gttccagtta
     8881 cagcgctctg acctgcagtg gacggagatt ttcccttact actaaagccc gcgtccggat
     8941 tacccgctgt agtcgtgctt gctacgctgc gtgactggtc cgcaatgaga cgctttgcgc
     9001 gctttcggca ggcgtccgag cagtagattt tggggcgctt cccggggatg tggacgatcg
     9061 gggtgccgca gtggcacttc ggtccggcgg ggcgcggtgg tgtcgacgcg ctgttctctc
     9121 gtacgctcgt cacagagcaa acgtcctcac tcggcatgct gcgccggttc gggggcggcg
     9181 agcccgggag gccaatcccg ggctcgtgcc atttctgggt cctgttgatc ctggcattgg
     9241 tgtggccgtt cattgcccct gctcgctcct gacgcgccga tagacgtccg atacgcccgg
     9301 tgctggtggg atttgatagg tcggaagaag ccccgcccgg gcctgggcgg ggcttcctgt
     9361 gcgtcaggac ctcctcgtcg tgagcctctt cggcctatgg acggagtgac ctcgtgatcc
     9421 gttacagccg cgcgcgctcg cgtagagcgg tctcatcagt tccacgaacg gtcctcttcg
     9481 cagatcaggg cgttggggcg gagtctcacc aaggactacg tctgctggcg atttccgtta
     9541 caccccgggc ggtggccggc gcacacgcgc gcccgcgttg ggcagtgcag aaagtgcaga
     9601 aacctaggcg ctgatggtcc aggtccacgg ttcgtcgtcg gcggcggcgc gggcggcggc
     9661 gtcggccagg gcgcgggcga gaccggctac ggcgggcttg atgcgccggt tgcgggcgac
     9721 cttgagcagc tagtatgcag gtcgacggat cttttccgct gcataaccct gcttcggggt
     9781 cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca
     9841 aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga
     9901 agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt
     9961 gctcaacggg aatcctgctc tgcgaggctg gccggctacc gccggcgtaa cagatgaggg
    10021 caagcggatg gctgatgaaa ccaagccaac caggaagggc agcccaccta tcaaggtgta
    10081 ctgccttcca gacgaacgaa gagcgattga ggaaaaggcg gcggcggccg gcatgagcct
    10141 gtcggcctac ctgctggccg tcggccaggg ctacaaaatc acgggcgtcg tggactatga
    10201 gcacgtccgc gagctg
//