LOCUS       sequence-416632-       12265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  improved pCRISPR-cBEST plasmid with optimized sgRNA cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12265)
  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 12265)
  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-PermE
FEATURES             Location/Qualifiers
     source          1..12265
                     /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        3137..3212
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(3382..3416)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             3584..4270
                     /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(4319..4342,4343..4345,4346..8419)
                     /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             8432..8683
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     terminator      complement(8864..8912)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             10627..12069
                     /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 agcttgcggt cgatcttgac ggctggcgag aggtgcgggg aggatctgac cgacgcggtc
     3061 cacacgtggc accgcgatgc tgttgtgggc acaatcgtgc cggttggtag gatcgacggc
     3121 tagcgtcaga tccatggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     3181 atcaacttga aaaagtggca ccgagtcggt gcttttttta cgtaggcgtc agctacgatg
     3241 ttccggggac tgctgatccg gtcagcaggt ggaagaggga ctggattcca aagttctcaa
     3301 tgctgcttgc tgttcttgaa tggggggtcg ttgacgacga catggctcga ttggcgcgac
     3361 aagttgctgc gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     3421 tccagatgga gttctgaggt cattactgga ccggatcggg gatctgggct gagggagccg
     3481 acggcacgcg gcggctcacg gcgtggcacg cggaacgtcc gggcttgcac ctcacgtcac
     3541 gtgaggaggc agcgtggacg gcgtcagaga agggagcgga catatgtcct ccgagaccgg
     3601 ccccgtggcc gtggacccga ccctgcgtcg ccgcatcgag ccgcacgagt tcgaggtctt
     3661 cttcgacccg cgcgagctgc gcaaggagac ctgcctgctg tacgagatca actggggcgg
     3721 ccggcactcc atctggcgcc acacctcgca gaacaccaac aagcacgtcg aggtcaactt
     3781 catcgagaag ttcaccaccg agcggtactt ctgcccgaac acccgctgct ccatcacctg
     3841 gttcctgtcc tggtccccct gcggcgagtg ctcccgggcc atcaccgagt tcctgtcccg
     3901 ctacccgcac gtcaccctgt tcatctacat cgcccggctg taccaccacg ccgacccgcg
     3961 caaccggcag ggcctgcgcg acctgatctc ctccggcgtc accatccaga tcatgaccga
     4021 gcaggagtcc ggctactgct ggcggaactt cgtcaactac tccccgtcca acgaggccca
     4081 ctggccccgc tacccgcacc tgtgggtccg cctgtacgtc ctggagctgt actgcatcat
     4141 cctgggcctg ccgccctgcc tgaacatcct gcgtcgcaag cagccgcagc tgaccttctt
     4201 caccatcgcc ctccagtcct gccactacca gcgcctgccg ccccacatcc tgtgggccac
     4261 cggcctgaag tccggctccg agaccccggg cacctcggag tccgccaccc cggagtccga
     4321 caagaagtac tccatcggcc tcgcgatcgg caccaactcc gtgggctggg cggtcatcac
     4381 cgacgagtac aaggtcccct ccaagaagtt caaggtcctg ggcaacaccg accggcactc
     4441 gatcaagaag aacctgatcg gcgccctgct cttcgacagc ggcgagaccg ccgaggcgac
     4501 ccgcctgaag cggaccgcgc gtcgccgcta cacccggcgc aagaaccgca tctgctacct
     4561 gcaggaaatc ttctccaacg agatggccaa ggtggacgac tcgttcttcc accgcctgga
     4621 ggagagcttc ctggtggagg aggacaagaa gcacgagcgc cacccgatct tcggcaacat
     4681 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctcc gcaagaagct
     4741 ggtggactcg accgacaagg cggacctgcg gctcatctac ctggccctcg cgcacatgat
     4801 caagttccgc ggccacttcc tcatcgaggg cgacctgaac ccggacaact ccgacgtgga
     4861 caagctcttc atccagctgg tgcagaccta caaccagctg ttcgaggaga accccatcaa
     4921 cgccagcggc gtggacgcca aggcgatcct ctccgcgcgc ctgagcaagt cccggcgcct
     4981 ggagaacctc atcgcccagc tgccgggcga gaagaagaac ggcctcttcg gcaacctgat
     5041 cgcgctgtcg ctcggcctga cccccaactt caagagcaac ttcgacctgg ccgaggacgc
     5101 gaagctccag ctgtccaagg acacctacga cgacgacctg gacaacctgc tcgcccagat
     5161 cggcgaccag tacgcggacc tcttcctggc cgcgaagaac ctctcggacg ccatcctgct
     5221 cagcgacatc ctgcgggtca acaccgagat caccaaggcc ccgctgtcgg cgagcatgat
     5281 caagcggtac gacgagcacc accaggacct gaccctgctc aaggccctcg tgcgccagca
     5341 gctgcccgag aagtacaagg aaatcttctt cgaccagtcc aagaacggct acgccggcta
     5401 catcgacggc ggcgcgtcgc aggaggagtt ctacaagttc atcaagccga tcctggagaa
     5461 gatggacggc accgaggagc tgctcgtcaa gctgaaccgc gaggacctgc tccgcaagca
     5521 gcggaccttc gacaacggct ccatcccgca ccagatccac ctgggcgagc tccacgccat
     5581 cctccggcgc caggaggact tctacccctt cctgaaggac aaccgcgaga agatcgagaa
     5641 gatcctgacc ttccgcatcc cgtactacgt cggccccctg gcccgcggca actcccggtt
     5701 cgcgtggatg acccggaagt cggaggagac catcaccccg tggaacttcg aggaggtcgt
     5761 ggacaagggc gcgtccgcgc agtcgttcat cgagcgcatg accaacttcg acaagaacct
     5821 cccgaacgag aaggtcctgc ccaagcactc cctgctctac gagtacttca ccgtgtacaa
     5881 cgagctgacc aaggtcaagt acgtgaccga gggcatgcgg aagccggcct tcctgtcggg
     5941 cgagcagaag aaggcgatcg tggacctgct cttcaagacc aaccgcaagg tcaccgtgaa
     6001 gcagctgaag gaggactact tcaagaagat cgagtgcttc gactccgtcg agatcagcgg
     6061 cgtggaggac cgcttcaacg cctccctggg cacctaccac gacctgctca agatcatcaa
     6121 ggacaaggac ttcctcgaca acgaggagaa cgaggacatc ctggaggaca tcgtcctcac
     6181 cctgaccctc ttcgaggacc gcgagatgat cgaggagcgg ctcaagacct acgcccacct
     6241 gttcgacgac aaggtgatga agcagctgaa gcgtcgccgc tacaccggct ggggccgcct
     6301 ctcccggaag ctgatcaacg gcatccggga caagcagagc ggcaagacca tcctggactt
     6361 cctcaagtcc gacggcttcg ccaaccgcaa cttcatgcag ctcatccacg acgacagcct
     6421 gaccttcaag gaggacatcc agaaggccca ggtctcgggc cagggcgaca gcctccacga
     6481 gcacatcgcc aacctggcgg gctccccggc gatcaagaag ggcatcctcc agaccgtcaa
     6541 ggtcgtggac gagctggtca aggtgatggg ccgccacaag cccgagaaca tcgtgatcga
     6601 gatggcccgg gagaaccaga ccacccagaa gggccagaag aactcgcgcg agcggatgaa
     6661 gcggatcgag gagggcatca aggagctcgg cagccagatc ctgaaggagc acccggtcga
     6721 gaacacccag ctgcagaacg agaagctgta cctctactac ctgcagaacg gccgcgacat
     6781 gtacgtggac caggagctcg acatcaaccg gctgtccgac tacgacgtgg accacatcgt
     6841 gccgcagtcc ttcctgaagg acgactcgat cgacaacaag gtcctgaccc gctcggacaa
     6901 gaaccggggc aagtccgaca acgtgccctc ggaggaggtc gtgaagaaga tgaagaacta
     6961 ctggcgccag ctgctcaacg ccaagctcat cacccagcgc aagttcgaca acctgaccaa
     7021 ggccgagcgg ggcggcctga gcgagctcga caaggcgggc ttcatcaagc gccagctggt
     7081 cgagacccgg cagatcacca agcacgtggc ccagatcctg gactcccgga tgaacaccaa
     7141 gtacgacgag aacgacaagc tgatccgcga ggtcaaggtg atcaccctca agagcaagct
     7201 ggtctccgac ttccgcaagg acttccagtt ctacaaggtc cgggagatca acaactacca
     7261 ccacgcccac gacgcgtacc tgaacgccgt cgtgggcacc gcgctgatca agaagtaccc
     7321 gaagctggag tccgagttcg tctacggcga ctacaaggtc tacgacgtgc gcaagatgat
     7381 cgccaagagc gagcaggaga tcggcaaggc caccgcgaag tacttcttct actccaacat
     7441 catgaacttc ttcaagaccg agatcaccct ggccaacggc gagatccgca agcggcccct
     7501 gatcgagacc aacggcgaga ccggcgagat cgtctgggac aagggccgcg acttcgccac
     7561 cgtccggaag gtgctgtcga tgccgcaggt caacatcgtg aagaagaccg aggtgcagac
     7621 cggcggcttc agcaaggagt ccatcctccc caagcgcaac agcgacaagc tgatcgcccg
     7681 gaagaaggac tgggacccga agaagtacgg cggcttcgac agccccaccg tcgcctactc
     7741 cgtgctggtc gtggcgaagg tcgagaaggg caagagcaag aagctgaagt ccgtgaagga
     7801 gctgctcggc atcaccatca tggagcgctc ctcgttcgag aagaacccga tcgacttcct
     7861 ggaggccaag ggctacaagg aggtcaagaa ggacctcatc atcaagctgc ccaagtacag
     7921 cctgttcgag ctggagaacg gccgcaagcg gatgctcgcc tccgcgggcg agctgcagaa
     7981 gggcaacgag ctggccctcc cgtcgaagta cgtcaacttc ctgtacctcg cgtcccacta
     8041 cgagaagctg aagggctcgc ccgaggacaa cgagcagaag cagctcttcg tggagcagca
     8101 caagcactac ctggacgaga tcatcgagca gatcagcgag ttcagcaagc gcgtcatcct
     8161 ggccgacgcg aacctcgaca aggtgctgtc cgcctacaac aagcaccgcg acaagccgat
     8221 ccgggagcag gcggagaaca tcatccacct gttcaccctc accaacctgg gcgcccccgc
     8281 cgcgttcaag tacttcgaca ccaccatcga ccgcaagcgg tacacctcca ccaaggaggt
     8341 cctcgacgcg accctgatcc accagagcat caccggcctg tacgagaccc gcatcgacct
     8401 gtcccagctc ggcggcgact ccggcggctc caccaacctg tccgacatca tcgagaagga
     8461 gaccggcaag cagctggtca tccaggagtc catcctgatg ctgccggagg aagtcgagga
     8521 agtcatcggc aacaagcccg agtccgacat cctggtccac accgcctacg acgagtccac
     8581 cgacgagaac gtcatgctgc tgacctcgga cgcccccgag tacaagccct gggccctggt
     8641 catccaggac tccaacggcg agaacaagat caagatgctg tgatctagag gatccccggg
     8701 taccgagctc gaattcccca gatctaaagt tttgtcgtct ttccagacgt tagtaaatga
     8761 attttctgta tgaggttttg ctaaacaact ttcaacagtt tcagcggagt gagaatagaa
     8821 aggaacaact aaaggaattg cgaataataa ttttttcacg ttgaaaatct ccaaaaaaaa
     8881 aggctccaaa aggagccttt aattgtatcg gtttatcagc ttgctttcga ggtgaatttc
     8941 ttaaacagct tgataccgat agttgcgccg acaatgacaa caaccatcgc ccacgcataa
     9001 ccgatatatt cggtcgctga ggcttgcagg gagtcaaagg ccgcttttgc gggatctcgt
     9061 cgaaggcact agaggcctat atgcggggat cgaccgcgcg ggtcccggac ggggaagagc
     9121 ggggagcttt gccagagagc gacgacttcc ccttgcgttg gtgattgccg gtcagggcag
     9181 ccatccgcca tcgtcgcgta gggtgtcaca ccccaggaat cgcgtcactg aacacagcag
     9241 ccggtaggac gaccatgact gagttggaca ccatcgcaaa tccgtccgat cccgcggtgc
     9301 agcggatcat cgatgtcacc aagccgtcgc gatccaacat aaagacaacg ttgatcgagg
     9361 acgtcgagcc cctcatgcac agcatcgcgg ccggggtgga gttcatcgag gtctacggca
     9421 gcgacagcag tccttttcca tctgagttgc tggatctgtg cgggcggcag aacataccgg
     9481 tccgcctcat cgactcctcg atcgtcaacc agttgttcaa gggggagcgg aaggccaaga
     9541 cattcggcat cgcccgcgtc cctcgcccgg ccaggttcgg cgatatcgcg agccggcgtg
     9601 gggacgtcgt cgttctcgac ggggtgaaga tcgtcgggaa catcggcgcg atagtacgca
     9661 cgtcgctcgc gctcggagcg tcggggatca tcctggtcga cagtgacatc accagcatcg
     9721 cggaccggcg tctccaaagg gccagccgag gttacgtctt ctcccttccc gtcgttctct
     9781 ccggtcgcga ggaggccatc gccttcattc gggacagcgg tatgcagctg atgacgctca
     9841 aggcggatgg cgacatttcc gtgaaggaac tcggggacaa tccggatcgg ctggccttgc
     9901 tgttcggcag cgaaaagggt gggccttccg acctgttcga ggaggcgtct tccgcctcgg
     9961 tttccatccc catgatgagc cagaccgagt ctctcaacgt ttccgtttcc ctcggaatcg
    10021 cgctgcacga gaggatcgac aggaatctcg cggccaaccg ataagcgcct ctgttcctcg
    10081 gacgctcggt tgctcgacct cgattcgtca gtgatgatca acaggaccca gaaatggcac
    10141 gagcccggga ttggcctccc gggctcgccg cccccgaacc ggcgcagcat gccgagtgag
    10201 gacgtttgct ctgtgacgag cgtacgagag aacagcgcgt cgacaccacc gcgccccgcc
    10261 ggaccgaagt gccactgcgg caccccgatc gtccacatcc ccgggaagcg ccccaaaatc
    10321 tactgctcgg acgcctgccg aaagcgcgca aagcgtctca ttgcggacca gtcacgcagc
    10381 gtagcaagca cgactacagc gggtaatccg gacgcgggct ttagtagtaa gggaaaatct
    10441 ccgtccactg caggtcagag cgctgtaact ggaacgggtt ctaccccaaa cgagggtgcc
    10501 gcgacagccg ggcgggagga cccgcgcgac gagcggttcg cccggcggga ccggcaccag
    10561 acggtgtcgc tgaacgaggc gttcaagggc tgcgggaccc ggctcactgc cgggacggcc
    10621 gagctgatgt ggaagccggg cgaggcgacg tggggcaaca cgtgccgctg caacaacgtc
    10681 cacacctgcc cgtggtgcat gagccgcatc ctggccgtac ggggctcgaa cgtgcagctc
    10741 gccgccgacg gtctcgcgga cgccgggtac gggctgcacc ttggcacgaa cacgttgcgc
    10801 cacttcgagc gcatggcgtt cggcaccgtc cgcaagggaa tgcggcacgg cctggtcgcg
    10861 gtcttgcacg acggctggaa gggcgcgtac ggctcgtccg gccgccgctg gcgcacgatg
    10921 cgcgacgact tcgggatcat cggctacgag cgggccttcg aggacacctt cggctggggc
    10981 tccggctggc acctgcactg gcacacgctc tgggtcaccc gcgaggtgct cggcccggac
    11041 gcccaggccg cgttccgcga cgcactcgcc ggagcctggg ccgccggggt cgaatccgcg
    11101 ggcggctaca ccgtcagcga gacgtgcgac cggcccggtt gctcctgtga gggcaagggc
    11161 cacggcaccg acgtgcgccc actcaacggc gcggacgccg ctgacggcga tgcgggcaag
    11221 caggcccgct acctctacaa ggacggcgac aagaccaagg gcggcgtcgc caagatcggc
    11281 ctggagctgg ccggacagaa cttcaaggcc ggtcgcggcg acgaccgcat gggaccgctc
    11341 gacctgggcg acgcggcggc cgccgagctg cagcggcttc gtcggcccgg cccgttcgtc
    11401 gagaagtacc gcgagcgcga gttcggcgtc ttccaggtcc gcaagcacta ccggtcgcag
    11461 aacctgaacc ggctgatcaa ggaactcggc atccagcagg acgtccgcac cgaggaggag
    11521 atcaccgacg acaccgaggg cctggtcgcc atcgcggtca tcccggccta catctggtac
    11581 cgatacatcg cccgcgtcgc cggacgtcgc ctcgacctca tcaaggtcgc cgagacgtac
    11641 ggcctaccgg gcgtccggcg gctcgtcgag tcctggggcc tggtgtgggg caaggacgtc
    11701 ctggacccgc ccgccccgga agcgccggcc gcgccgggcg accttgacgc tgaccagatg
    11761 aggttcgagg tcatgtccga ggaggaagct gctttccgcg aggcgcgccg gaaggccaac
    11821 gaggcccgca cggaggagct ggccgcttcg ctcgaccgag tgcgtcagcc gaagaaggag
    11881 gcgatccgcc cgacgatcag ccttcggaag cgcctgaagc cgaagccggt caccgtcgac
    11941 gtcaagactc ccccgcccgg cgccgcgtcg cccgtgtgcc gccgctgcaa gggcaagctc
    12001 gccccggtac tccagccctg gggccagcac cccggagact gcctccgcgt cgacacagcg
    12061 gtcgcgtagg gccgctgagg gaccgccatg agaccgccac cccggacatc cgggtggcgg
    12121 tctcatggcg gtcccggcgc gaaagccgag ggtctgcctg ccgtgaggtg gccggcggac
    12181 ctgggttact gctggaatcc cgcgcccagt ccgagcgtcc gccggcgctt ttcacctggc
    12241 tccgggtgct cgtcgatgag cacca
//
