LOCUS       sequence-416625-       12234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  improved pCRISPR-cBEST plasmid with optimized sgRNA cloning site and
            expression of the sgRNA from the SF14P promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12234)
  AUTHORS   Maleckis M, Wibowo M, Gren T, Jarmusch SA, Sterndorff EB, Booth T, 
            Henriksen NNSE, Whitford CM, Jiang X, Jorgensen TS, Ding L, Weber T
  TITLE     Biosynthesis of the Azoxy Compound Azodyrecin from Streptomyces 
            mirabilis P8-A2.
  JOURNAL   ACS Chem Biol. 2024 Feb 10. doi: 10.1021/acschembio.3c00632.
  PUBMED    38340355
REFERENCE   2  (bases 1 to 12234)
  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 Chem 
            Biol. 2024 Feb 10. doi: 10.1021/acschembio.3c00632."
COMMENT     Sequence Label: pCRISPR-cBEST-v2-SF14P
FEATURES             Location/Qualifiers
     source          1..12234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            466..575
                     /note="incP origin of transfer"
     CDS             891..1691
                     /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
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDSFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELLAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      1993..2580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2481..2500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2734..2751
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2868..2889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2904..2909,2910..2927,2928..2934)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2942..2958
                     /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)."
     primer_bind     2947..2969
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2966..2982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2966..2982
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        3106..3181
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(3351..3385)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             3553..4239
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(4288..4311,4312..4314,4315..8388)
                     /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"
     CDS             8401..8652
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     terminator      complement(8833..8881)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             10596..12038
                     /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 ggccgtcgac gacgacgcgt ccgcctgcct gcttttggcc ggcgggccgg gccgcggttg
       61 tccgccgggt cggaccggtc ccttggtcga ccgatgcctc ggccacgacg atgctgggcc
      121 cgaagcggta cgcgacccgt gccgggccgc agcggatcgc ggcagcagga gctccccgcc
      181 cgccacggct ggctggatgg tcgcctcacg cgcccgcagg ggtgacggcg tgggtgcggc
      241 ggtcagcccc gtgatcgggc gtcagacggc cgctgaggga ccgcgaccac ccctcccttc
      301 ccgcctgccg tctttcaccg cccggccggt ccgcagacgc cgtacggcca ggccacggag
      361 gattacgggc agatggcgcc cacccggcac ccgggctggg gcaccctgga cagcacgaag
      421 gtccgcccca ctcaccgggg cgggccttcg cgttgagcta gagttcccgc cagcctcgca
      481 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
      541 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
      601 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
      661 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaat gcagctcacg
      721 gtaactgatg ccgtatttgc agtaccagcg tacggcccac agaatgatgt cacgctgaaa
      781 atgccggcct ttgaatgggt tcatgtgcag ctccataagc aaaaggggat gataagttta
      841 tcaccaccga ctatttgcaa cagtgccgtt gatcgtgcta tgatcgactg atgtcatcag
      901 cggtggagtg caatgtcgtg caatacgaat ggcgaaaagc cgagctcatc ggtcagcttc
      961 tcaaccttgg ggttaccccc ggcggtgtgc tgctggtcca cagctccttc cgtagcgtcc
     1021 ggcccctcga agatgggcca cttggactga tcgaggccct gcgtgctgcg ctgggtccgg
     1081 gagggacgct cgtcatgccc tcgtggtcag gtctggacga cgagccgttc gatcctgcca
     1141 cgtcgcccgt tacaccggac cttggagttg tctctgactc attctggcgc ctgccaaatg
     1201 taaagcgcag cgcccatcca tttgcctttg cggcagcggg gccacaggca gagcagatca
     1261 tctctgatcc attgcccctg ccacctcact cgcctgcaag cccggtcgcc cgtgtccatg
     1321 aactcgatgg gcaggtactt ctcctcggcg tgggacacga tgccaacacg acgctgcatc
     1381 ttgccgagtt gttggcaaag gttccctatg gggtgccgag acactgcacc attcttcagg
     1441 atggcaagtt ggtacgcgtc gattatctcg agaatgacca ctgctgtgag cgctttgcct
     1501 tggcggacag gtggctcaag gagaagagcc ttcagaagga aggtccagtc ggtcatgcct
     1561 ttgctcggtt gatccgctcc cgcgacattg tggcgacagc cctgggtcaa ctgggccgag
     1621 atccgttgat cttcctgcat ccgccagagg cgggatgcga agaatgcgat gccgctcgcc
     1681 agtcgattgg ctgagctcat aagttcctat tccgaagttc ctattcgaaa tgaccgacca
     1741 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     1801 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     1861 gctggagttc ttcgcccacc ccaaaaggat ctaggtgaag atcctttttg ataatctcat
     1921 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1981 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2041 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2101 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2161 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2221 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2281 gttaccggat aaggcgcagc ggtcgggctg aacggggggt cgtgcacaca gcccagcttg
     2341 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2401 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2461 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2521 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2581 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2641 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2701 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2761 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2821 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     2881 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     2941 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattgtaca
     3001 agcttcctat ccaggagata ttatgagtta cgtagaccta cgccttgacc ttgatgaggc
     3061 ggcgtgagct acaatcaata ctcgattgct agcgtcagat ccatggtttt agagctagaa
     3121 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     3181 ctttttttac gtaggcgtca gctacgatgt tccggggact gctgatccgg tcagcaggtg
     3241 gaagagggac tggattccaa agttctcaat gctgcttgct gttcttgaat ggggggtcgt
     3301 tgacgacgac atggctcgat tggcgcgaca agttgctgcg attctcacca ataaaaaacg
     3361 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggac
     3421 cggatcgggg atctgggctg agggagccga cggcacgcgg cggctcacgg cgtggcacgc
     3481 ggaacgtccg ggcttgcacc tcacgtcacg tgaggaggca gcgtggacgg cgtcagagaa
     3541 gggagcggac atatgtcctc cgagaccggc cccgtggccg tggacccgac cctgcgtcgc
     3601 cgcatcgagc cgcacgagtt cgaggtcttc ttcgacccgc gcgagctgcg caaggagacc
     3661 tgcctgctgt acgagatcaa ctggggcggc cggcactcca tctggcgcca cacctcgcag
     3721 aacaccaaca agcacgtcga ggtcaacttc atcgagaagt tcaccaccga gcggtacttc
     3781 tgcccgaaca cccgctgctc catcacctgg ttcctgtcct ggtccccctg cggcgagtgc
     3841 tcccgggcca tcaccgagtt cctgtcccgc tacccgcacg tcaccctgtt catctacatc
     3901 gcccggctgt accaccacgc cgacccgcgc aaccggcagg gcctgcgcga cctgatctcc
     3961 tccggcgtca ccatccagat catgaccgag caggagtccg gctactgctg gcggaacttc
     4021 gtcaactact ccccgtccaa cgaggcccac tggccccgct acccgcacct gtgggtccgc
     4081 ctgtacgtcc tggagctgta ctgcatcatc ctgggcctgc cgccctgcct gaacatcctg
     4141 cgtcgcaagc agccgcagct gaccttcttc accatcgccc tccagtcctg ccactaccag
     4201 cgcctgccgc cccacatcct gtgggccacc ggcctgaagt ccggctccga gaccccgggc
     4261 acctcggagt ccgccacccc ggagtccgac aagaagtact ccatcggcct cgcgatcggc
     4321 accaactccg tgggctgggc ggtcatcacc gacgagtaca aggtcccctc caagaagttc
     4381 aaggtcctgg gcaacaccga ccggcactcg atcaagaaga acctgatcgg cgccctgctc
     4441 ttcgacagcg gcgagaccgc cgaggcgacc cgcctgaagc ggaccgcgcg tcgccgctac
     4501 acccggcgca agaaccgcat ctgctacctg caggaaatct tctccaacga gatggccaag
     4561 gtggacgact cgttcttcca ccgcctggag gagagcttcc tggtggagga ggacaagaag
     4621 cacgagcgcc acccgatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     4681 cccaccatct accacctccg caagaagctg gtggactcga ccgacaaggc ggacctgcgg
     4741 ctcatctacc tggccctcgc gcacatgatc aagttccgcg gccacttcct catcgagggc
     4801 gacctgaacc cggacaactc cgacgtggac aagctcttca tccagctggt gcagacctac
     4861 aaccagctgt tcgaggagaa ccccatcaac gccagcggcg tggacgccaa ggcgatcctc
     4921 tccgcgcgcc tgagcaagtc ccggcgcctg gagaacctca tcgcccagct gccgggcgag
     4981 aagaagaacg gcctcttcgg caacctgatc gcgctgtcgc tcggcctgac ccccaacttc
     5041 aagagcaact tcgacctggc cgaggacgcg aagctccagc tgtccaagga cacctacgac
     5101 gacgacctgg acaacctgct cgcccagatc ggcgaccagt acgcggacct cttcctggcc
     5161 gcgaagaacc tctcggacgc catcctgctc agcgacatcc tgcgggtcaa caccgagatc
     5221 accaaggccc cgctgtcggc gagcatgatc aagcggtacg acgagcacca ccaggacctg
     5281 accctgctca aggccctcgt gcgccagcag ctgcccgaga agtacaagga aatcttcttc
     5341 gaccagtcca agaacggcta cgccggctac atcgacggcg gcgcgtcgca ggaggagttc
     5401 tacaagttca tcaagccgat cctggagaag atggacggca ccgaggagct gctcgtcaag
     5461 ctgaaccgcg aggacctgct ccgcaagcag cggaccttcg acaacggctc catcccgcac
     5521 cagatccacc tgggcgagct ccacgccatc ctccggcgcc aggaggactt ctaccccttc
     5581 ctgaaggaca accgcgagaa gatcgagaag atcctgacct tccgcatccc gtactacgtc
     5641 ggccccctgg cccgcggcaa ctcccggttc gcgtggatga cccggaagtc ggaggagacc
     5701 atcaccccgt ggaacttcga ggaggtcgtg gacaagggcg cgtccgcgca gtcgttcatc
     5761 gagcgcatga ccaacttcga caagaacctc ccgaacgaga aggtcctgcc caagcactcc
     5821 ctgctctacg agtacttcac cgtgtacaac gagctgacca aggtcaagta cgtgaccgag
     5881 ggcatgcgga agccggcctt cctgtcgggc gagcagaaga aggcgatcgt ggacctgctc
     5941 ttcaagacca accgcaaggt caccgtgaag cagctgaagg aggactactt caagaagatc
     6001 gagtgcttcg actccgtcga gatcagcggc gtggaggacc gcttcaacgc ctccctgggc
     6061 acctaccacg acctgctcaa gatcatcaag gacaaggact tcctcgacaa cgaggagaac
     6121 gaggacatcc tggaggacat cgtcctcacc ctgaccctct tcgaggaccg cgagatgatc
     6181 gaggagcggc tcaagaccta cgcccacctg ttcgacgaca aggtgatgaa gcagctgaag
     6241 cgtcgccgct acaccggctg gggccgcctc tcccggaagc tgatcaacgg catccgggac
     6301 aagcagagcg gcaagaccat cctggacttc ctcaagtccg acggcttcgc caaccgcaac
     6361 ttcatgcagc tcatccacga cgacagcctg accttcaagg aggacatcca gaaggcccag
     6421 gtctcgggcc agggcgacag cctccacgag cacatcgcca acctggcggg ctccccggcg
     6481 atcaagaagg gcatcctcca gaccgtcaag gtcgtggacg agctggtcaa ggtgatgggc
     6541 cgccacaagc ccgagaacat cgtgatcgag atggcccggg agaaccagac cacccagaag
     6601 ggccagaaga actcgcgcga gcggatgaag cggatcgagg agggcatcaa ggagctcggc
     6661 agccagatcc tgaaggagca cccggtcgag aacacccagc tgcagaacga gaagctgtac
     6721 ctctactacc tgcagaacgg ccgcgacatg tacgtggacc aggagctcga catcaaccgg
     6781 ctgtccgact acgacgtgga ccacatcgtg ccgcagtcct tcctgaagga cgactcgatc
     6841 gacaacaagg tcctgacccg ctcggacaag aaccggggca agtccgacaa cgtgccctcg
     6901 gaggaggtcg tgaagaagat gaagaactac tggcgccagc tgctcaacgc caagctcatc
     6961 acccagcgca agttcgacaa cctgaccaag gccgagcggg gcggcctgag cgagctcgac
     7021 aaggcgggct tcatcaagcg ccagctggtc gagacccggc agatcaccaa gcacgtggcc
     7081 cagatcctgg actcccggat gaacaccaag tacgacgaga acgacaagct gatccgcgag
     7141 gtcaaggtga tcaccctcaa gagcaagctg gtctccgact tccgcaagga cttccagttc
     7201 tacaaggtcc gggagatcaa caactaccac cacgcccacg acgcgtacct gaacgccgtc
     7261 gtgggcaccg cgctgatcaa gaagtacccg aagctggagt ccgagttcgt ctacggcgac
     7321 tacaaggtct acgacgtgcg caagatgatc gccaagagcg agcaggagat cggcaaggcc
     7381 accgcgaagt acttcttcta ctccaacatc atgaacttct tcaagaccga gatcaccctg
     7441 gccaacggcg agatccgcaa gcggcccctg atcgagacca acggcgagac cggcgagatc
     7501 gtctgggaca agggccgcga cttcgccacc gtccggaagg tgctgtcgat gccgcaggtc
     7561 aacatcgtga agaagaccga ggtgcagacc ggcggcttca gcaaggagtc catcctcccc
     7621 aagcgcaaca gcgacaagct gatcgcccgg aagaaggact gggacccgaa gaagtacggc
     7681 ggcttcgaca gccccaccgt cgcctactcc gtgctggtcg tggcgaaggt cgagaagggc
     7741 aagagcaaga agctgaagtc cgtgaaggag ctgctcggca tcaccatcat ggagcgctcc
     7801 tcgttcgaga agaacccgat cgacttcctg gaggccaagg gctacaagga ggtcaagaag
     7861 gacctcatca tcaagctgcc caagtacagc ctgttcgagc tggagaacgg ccgcaagcgg
     7921 atgctcgcct ccgcgggcga gctgcagaag ggcaacgagc tggccctccc gtcgaagtac
     7981 gtcaacttcc tgtacctcgc gtcccactac gagaagctga agggctcgcc cgaggacaac
     8041 gagcagaagc agctcttcgt ggagcagcac aagcactacc tggacgagat catcgagcag
     8101 atcagcgagt tcagcaagcg cgtcatcctg gccgacgcga acctcgacaa ggtgctgtcc
     8161 gcctacaaca agcaccgcga caagccgatc cgggagcagg cggagaacat catccacctg
     8221 ttcaccctca ccaacctggg cgcccccgcc gcgttcaagt acttcgacac caccatcgac
     8281 cgcaagcggt acacctccac caaggaggtc ctcgacgcga ccctgatcca ccagagcatc
     8341 accggcctgt acgagacccg catcgacctg tcccagctcg gcggcgactc cggcggctcc
     8401 accaacctgt ccgacatcat cgagaaggag accggcaagc agctggtcat ccaggagtcc
     8461 atcctgatgc tgccggagga agtcgaggaa gtcatcggca acaagcccga gtccgacatc
     8521 ctggtccaca ccgcctacga cgagtccacc gacgagaacg tcatgctgct gacctcggac
     8581 gcccccgagt acaagccctg ggccctggtc atccaggact ccaacggcga gaacaagatc
     8641 aagatgctgt gatctagagg atccccgggt accgagctcg aattccccag atctaaagtt
     8701 ttgtcgtctt tccagacgtt agtaaatgaa ttttctgtat gaggttttgc taaacaactt
     8761 tcaacagttt cagcggagtg agaatagaaa ggaacaacta aaggaattgc gaataataat
     8821 tttttcacgt tgaaaatctc caaaaaaaaa ggctccaaaa ggagccttta attgtatcgg
     8881 tttatcagct tgctttcgag gtgaatttct taaacagctt gataccgata gttgcgccga
     8941 caatgacaac aaccatcgcc cacgcataac cgatatattc ggtcgctgag gcttgcaggg
     9001 agtcaaaggc cgcttttgcg ggatctcgtc gaaggcacta gaggcctata tgcggggatc
     9061 gaccgcgcgg gtcccggacg gggaagagcg gggagctttg ccagagagcg acgacttccc
     9121 cttgcgttgg tgattgccgg tcagggcagc catccgccat cgtcgcgtag ggtgtcacac
     9181 cccaggaatc gcgtcactga acacagcagc cggtaggacg accatgactg agttggacac
     9241 catcgcaaat ccgtccgatc ccgcggtgca gcggatcatc gatgtcacca agccgtcgcg
     9301 atccaacata aagacaacgt tgatcgagga cgtcgagccc ctcatgcaca gcatcgcggc
     9361 cggggtggag ttcatcgagg tctacggcag cgacagcagt ccttttccat ctgagttgct
     9421 ggatctgtgc gggcggcaga acataccggt ccgcctcatc gactcctcga tcgtcaacca
     9481 gttgttcaag ggggagcgga aggccaagac attcggcatc gcccgcgtcc ctcgcccggc
     9541 caggttcggc gatatcgcga gccggcgtgg ggacgtcgtc gttctcgacg gggtgaagat
     9601 cgtcgggaac atcggcgcga tagtacgcac gtcgctcgcg ctcggagcgt cggggatcat
     9661 cctggtcgac agtgacatca ccagcatcgc ggaccggcgt ctccaaaggg ccagccgagg
     9721 ttacgtcttc tcccttcccg tcgttctctc cggtcgcgag gaggccatcg ccttcattcg
     9781 ggacagcggt atgcagctga tgacgctcaa ggcggatggc gacatttccg tgaaggaact
     9841 cggggacaat ccggatcggc tggccttgct gttcggcagc gaaaagggtg ggccttccga
     9901 cctgttcgag gaggcgtctt ccgcctcggt ttccatcccc atgatgagcc agaccgagtc
     9961 tctcaacgtt tccgtttccc tcggaatcgc gctgcacgag aggatcgaca ggaatctcgc
    10021 ggccaaccga taagcgcctc tgttcctcgg acgctcggtt gctcgacctc gattcgtcag
    10081 tgatgatcaa caggacccag aaatggcacg agcccgggat tggcctcccg ggctcgccgc
    10141 ccccgaaccg gcgcagcatg ccgagtgagg acgtttgctc tgtgacgagc gtacgagaga
    10201 acagcgcgtc gacaccaccg cgccccgccg gaccgaagtg ccactgcggc accccgatcg
    10261 tccacatccc cgggaagcgc cccaaaatct actgctcgga cgcctgccga aagcgcgcaa
    10321 agcgtctcat tgcggaccag tcacgcagcg tagcaagcac gactacagcg ggtaatccgg
    10381 acgcgggctt tagtagtaag ggaaaatctc cgtccactgc aggtcagagc gctgtaactg
    10441 gaacgggttc taccccaaac gagggtgccg cgacagccgg gcgggaggac ccgcgcgacg
    10501 agcggttcgc ccggcgggac cggcaccaga cggtgtcgct gaacgaggcg ttcaagggct
    10561 gcgggacccg gctcactgcc gggacggccg agctgatgtg gaagccgggc gaggcgacgt
    10621 ggggcaacac gtgccgctgc aacaacgtcc acacctgccc gtggtgcatg agccgcatcc
    10681 tggccgtacg gggctcgaac gtgcagctcg ccgccgacgg tctcgcggac gccgggtacg
    10741 ggctgcacct tggcacgaac acgttgcgcc acttcgagcg catggcgttc ggcaccgtcc
    10801 gcaagggaat gcggcacggc ctggtcgcgg tcttgcacga cggctggaag ggcgcgtacg
    10861 gctcgtccgg ccgccgctgg cgcacgatgc gcgacgactt cgggatcatc ggctacgagc
    10921 gggccttcga ggacaccttc ggctggggct ccggctggca cctgcactgg cacacgctct
    10981 gggtcacccg cgaggtgctc ggcccggacg cccaggccgc gttccgcgac gcactcgccg
    11041 gagcctgggc cgccggggtc gaatccgcgg gcggctacac cgtcagcgag acgtgcgacc
    11101 ggcccggttg ctcctgtgag ggcaagggcc acggcaccga cgtgcgccca ctcaacggcg
    11161 cggacgccgc tgacggcgat gcgggcaagc aggcccgcta cctctacaag gacggcgaca
    11221 agaccaaggg cggcgtcgcc aagatcggcc tggagctggc cggacagaac ttcaaggccg
    11281 gtcgcggcga cgaccgcatg ggaccgctcg acctgggcga cgcggcggcc gccgagctgc
    11341 agcggcttcg tcggcccggc ccgttcgtcg agaagtaccg cgagcgcgag ttcggcgtct
    11401 tccaggtccg caagcactac cggtcgcaga acctgaaccg gctgatcaag gaactcggca
    11461 tccagcagga cgtccgcacc gaggaggaga tcaccgacga caccgagggc ctggtcgcca
    11521 tcgcggtcat cccggcctac atctggtacc gatacatcgc ccgcgtcgcc ggacgtcgcc
    11581 tcgacctcat caaggtcgcc gagacgtacg gcctaccggg cgtccggcgg ctcgtcgagt
    11641 cctggggcct ggtgtggggc aaggacgtcc tggacccgcc cgccccggaa gcgccggccg
    11701 cgccgggcga ccttgacgct gaccagatga ggttcgaggt catgtccgag gaggaagctg
    11761 ctttccgcga ggcgcgccgg aaggccaacg aggcccgcac ggaggagctg gccgcttcgc
    11821 tcgaccgagt gcgtcagccg aagaaggagg cgatccgccc gacgatcagc cttcggaagc
    11881 gcctgaagcc gaagccggtc accgtcgacg tcaagactcc cccgcccggc gccgcgtcgc
    11941 ccgtgtgccg ccgctgcaag ggcaagctcg ccccggtact ccagccctgg ggccagcacc
    12001 ccggagactg cctccgcgtc gacacagcgg tcgcgtaggg ccgctgaggg accgccatga
    12061 gaccgccacc ccggacatcc gggtggcggt ctcatggcgg tcccggcgcg aaagccgagg
    12121 gtctgcctgc cgtgaggtgg ccggcggacc tgggttactg ctggaatccc gcgcccagtc
    12181 cgagcgtccg ccggcgcttt tcacctggct ccgggtgctc gtcgatgagc acca
//
