LOCUS       sequence-416613-       13212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for multiplexed cytosine base editing in streptomycetes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13212)
  AUTHORS   Whitford CM, Gren T, Palazzotto E, Lee SY, Tong Y, Weber T
  TITLE     Systems Analysis of Highly Multiplexed CRISPR-Base Editing in 
            Streptomycetes.
  JOURNAL   ACS Synth Biol. 2023 Aug 18;12(8):2353-2366. doi: 
            10.1021/acssynbio.3c00188. Epub 2023 Jul 4.
  PUBMED    37402223
REFERENCE   2  (bases 1 to 13212)
  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"; doi: 
            "10.1021/acssynbio.3c00188"; journalName: "ACS Synth Biol"; date: 
            "2023-08-18- 18"; volume: "12"; issue: "8"; pages: "2353-2366"
COMMENT     Sequence Label: pCRISPR-mcBEST
FEATURES             Location/Qualifiers
     source          1..13212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(197..1639)
                     /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"
     CDS             3318..4031
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHRFSVYGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(3618..3637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3965..3986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      4038..4071
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(4073..4324)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             complement(join(4337..8410,8411..8413,8414..8437))
                     /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             complement(8486..9172)
                     /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"
     terminator      9340..9374
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(9535..10098)
                     /codon_start=1
                     /product="CRISPR-associated endoribonuclease responsible 
                     for pre-crRNA processing in Pseudomonas aeruginosa 
                     (Haurwitz et al., 2010)"
                     /label=Csy4
                     /translation="MDHYLDIRLRPDPEFPPAQLMSVLFGKLHQALVAQGGDRIGVSFP
                     DLDESRSRLGERLRIHASADDLRALLARPWLEGLRDHLQFGEPAVVPHPTPYRQVSRVQ
                     AKSNPERLRRRLMRRHDLSEEEARKRIPDTVARALDLPFVTLRSQSTGQHFRLFIRHGP
                     LQVTAEEGGFTCYGLSKGGSVPWF"
     primer_bind     complement(10231..10247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10231..10247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10244..10266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10255..10271
                     /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(10279..10285,10286..10303,10304..10309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10324..10345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10462..10479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10633..11220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10713..10732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11522..12322)
                     /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"
     oriT            12638..12747
                     /note="incP origin of transfer"
ORIGIN
        1 tggtgctcat cgacgagcac ccggagccag gtgaaaagcg ccggcggacg ctcggactgg
       61 gcgcgggatt ccagcagtaa cccaggtccg ccggccacct cacggcaggc agaccctcgg
      121 ctttcgcgcc gggaccgcca tgagaccgcc acccggatgt ccggggtggc ggtctcatgg
      181 cggtccctca gcggccctac gcgaccgctg tgtcgacgcg gaggcagtct ccggggtgct
      241 ggccccaggg ctggagtacc ggggcgagct tgcccttgca gcggcggcac acgggcgacg
      301 cggcgccggg cgggggagtc ttgacgtcga cggtgaccgg cttcggcttc aggcgcttcc
      361 gaaggctgat cgtcgggcgg atcgcctcct tcttcggctg acgcactcgg tcgagcgaag
      421 cggccagctc ctccgtgcgg gcctcgttgg ccttccggcg cgcctcgcgg aaagcagctt
      481 cctcctcgga catgacctcg aacctcatct ggtcagcgtc aaggtcgccc ggcgcggccg
      541 gcgcttccgg ggcgggcggg tccaggacgt ccttgcccca caccaggccc caggactcga
      601 cgagccgccg gacgcccggt aggccgtacg tctcggcgac cttgatgagg tcgaggcgac
      661 gtccggcgac gcgggcgatg tatcggtacc agatgtaggc cgggatgacc gcgatggcga
      721 ccaggccctc ggtgtcgtcg gtgatctcct cctcggtgcg gacgtcctgc tggatgccga
      781 gttccttgat cagccggttc aggttctgcg accggtagtg cttgcggacc tggaagacgc
      841 cgaactcgcg ctcgcggtac ttctcgacga acgggccggg ccgacgaagc cgctgcagct
      901 cggcggccgc cgcgtcgccc aggtcgagcg gtcccatgcg gtcgtcgccg cgaccggcct
      961 tgaagttctg tccggccagc tccaggccga tcttggcgac gccgcccttg gtcttgtcgc
     1021 cgtccttgta gaggtagcgg gcctgcttgc ccgcatcgcc gtcagcggcg tccgcgccgt
     1081 tgagtgggcg cacgtcggtg ccgtggccct tgccctcaca ggagcaaccg ggccggtcgc
     1141 acgtctcgct gacggtgtag ccgcccgcgg attcgacccc ggcggcccag gctccggcga
     1201 gtgcgtcgcg gaacgcggcc tgggcgtccg ggccgagcac ctcgcgggtg acccagagcg
     1261 tgtgccagtg caggtgccag ccggagcccc agccgaaggt gtcctcgaag gcccgctcgt
     1321 agccgatgat cccgaagtcg tcgcgcatcg tgcgccagcg gcggccggac gagccgtacg
     1381 cgcccttcca gccgtcgtgc aagaccgcga ccaggccgtg ccgcattccc ttgcggacgg
     1441 tgccgaacgc catgcgctcg aagtggcgca acgtgttcgt gccaaggtgc agcccgtacc
     1501 cggcgtccgc gagaccgtcg gcggcgagct gcacgttcga gccccgtacg gccaggatgc
     1561 ggctcatgca ccacgggcag gtgtggacgt tgttgcagcg gcacgtgttg ccccacgtcg
     1621 cctcgcccgg cttccacatc agctcggccg tcccggcagt gagccgggtc ccgcagccct
     1681 tgaacgcctc gttcagcgac accgtctggt gccggtcccg ccgggcgaac cgctcgtcgc
     1741 gcgggtcctc ccgcccggct gtcgcggcac cctcgtttgg ggtagaaccc gttccagtta
     1801 cagcgctctg acctgcagtg gacggagatt ttcccttact actaaagccc gcgtccggat
     1861 tacccgctgt agtcgtgctt gctacgctgc gtgactggtc cgcaatgaga cgctttgcgc
     1921 gctttcggca ggcgtccgag cagtagattt tggggcgctt cccggggatg tggacgatcg
     1981 gggtgccgca gtggcacttc ggtccggcgg ggcgcggtgg tgtcgacgcg ctgttctctc
     2041 gtacgctcgt cacagagcaa acgtcctcac tcggcatgct gcgccggttc gggggcggcg
     2101 agcccgggag gccaatcccg ggctcgtgcc atttctgggt cctgttgatc atcactgacg
     2161 aatcgaggtc gagcaaccga gcgtccgagg aacagaggcg cttatcggtt ggccgcgaga
     2221 ttcctgtcga tcctctcgtg cagcgcgatt ccgagggaaa cggaaacgtt gagagactcg
     2281 gtctggctca tcatggggat ggaaaccgag gcggaagacg cctcctcgaa caggtcggaa
     2341 ggcccaccct tttcgctgcc gaacagcaag gccagccgat ccggattgtc cccgagttcc
     2401 ttcacggaaa tgtcgccatc cgccttgagc gtcatcagct gcataccgct gtcccgaatg
     2461 aaggcgatgg cctcctcgcg accggagaga acgacgggaa gggagaagac gtaacctcgg
     2521 ctggcccttt ggagacgccg gtccgcgatg ctggtgatgt cactgtcgac caggatgatc
     2581 cccgacgctc cgagcgcgag cgacgtgcgt actatcgcgc cgatgttccc gacgatcttc
     2641 accccgtcga gaacgacgac gtccccacgc cggctcgcga tatcgccgaa cctggccggg
     2701 cgagggacgc gggcgatgcc gaatgtcttg gccttccgct cccccttgaa caactggttg
     2761 acgatcgagg agtcgatgag gcggaccggt atgttctgcc gcccgcacag atccagcaac
     2821 tcagatggaa aaggactgct gtcgctgccg tagacctcga tgaactccac cccggccgcg
     2881 atgctgtgca tgaggggctc gacgtcctcg atcaacgttg tctttatgtt ggatcgcgac
     2941 ggcttggtga catcgatgat ccgctgcacc gcgggatcgg acggatttgc gatggtgtcc
     3001 aactcagtca tggtcgtcct accggctgct gtgttcagtg acgcgattcc tggggtgtga
     3061 caccctacgc gacgatggcg gatggctgcc ctgaccggca atcaccaacg caaggggaag
     3121 tcgtcgctct ctggcaaagc tccccgctct tccccgtccg ggacccgcgc ggtcgatccc
     3181 cgcatatagg cctctagtgc cttcgacgag atctgttcac attcgaaccg tctctgcttt
     3241 gacaacatgc tgtgcggtgt tgtaaagtcg tggccatcta gtccatggcg agcaacggag
     3301 gtacggacat gcgatccagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     3361 tcgagctgga cggcgacgta aacggccaca ggttcagcgt gtacggcgag ggcgagggcg
     3421 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3481 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3541 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     3601 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     3661 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     3721 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     3781 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     3841 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     3901 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     3961 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     4021 tgtacaagtg agctagcttc agaacgctcg gttgccgccg ggcgtttttt agtcacagca
     4081 tcttgatctt gttctcgccg ttggagtcct ggatgaccag ggcccagggc ttgtactcgg
     4141 gggcgtccga ggtcagcagc atgacgttct cgtcggtgga ctcgtcgtag gcggtgtgga
     4201 ccaggatgtc ggactcgggc ttgttgccga tgacttcctc gacttcctcc ggcagcatca
     4261 ggatggactc ctggatgacc agctgcttgc cggtctcctt ctcgatgatg tcggacaggt
     4321 tggtggagcc gccggagtcg ccgccgagct gggacaggtc gatgcgggtc tcgtacaggc
     4381 cggtgatgct ctggtggatc agggtcgcgt cgaggacctc cttggtggag gtgtaccgct
     4441 tgcggtcgat ggtggtgtcg aagtacttga acgcggcggg ggcgcccagg ttggtgaggg
     4501 tgaacaggtg gatgatgttc tccgcctgct cccggatcgg cttgtcgcgg tgcttgttgt
     4561 aggcggacag caccttgtcg aggttcgcgt cggccaggat gacgcgcttg ctgaactcgc
     4621 tgatctgctc gatgatctcg tccaggtagt gcttgtgctg ctccacgaag agctgcttct
     4681 gctcgttgtc ctcgggcgag cccttcagct tctcgtagtg ggacgcgagg tacaggaagt
     4741 tgacgtactt cgacgggagg gccagctcgt tgcccttctg cagctcgccc gcggaggcga
     4801 gcatccgctt gcggccgttc tccagctcga acaggctgta cttgggcagc ttgatgatga
     4861 ggtccttctt gacctccttg tagcccttgg cctccaggaa gtcgatcggg ttcttctcga
     4921 acgaggagcg ctccatgatg gtgatgccga gcagctcctt cacggacttc agcttcttgc
     4981 tcttgccctt ctcgaccttc gccacgacca gcacggagta ggcgacggtg gggctgtcga
     5041 agccgccgta cttcttcggg tcccagtcct tcttccgggc gatcagcttg tcgctgttgc
     5101 gcttggggag gatggactcc ttgctgaagc cgccggtctg cacctcggtc ttcttcacga
     5161 tgttgacctg cggcatcgac agcaccttcc ggacggtggc gaagtcgcgg cccttgtccc
     5221 agacgatctc gccggtctcg ccgttggtct cgatcagggg ccgcttgcgg atctcgccgt
     5281 tggccagggt gatctcggtc ttgaagaagt tcatgatgtt ggagtagaag aagtacttcg
     5341 cggtggcctt gccgatctcc tgctcgctct tggcgatcat cttgcgcacg tcgtagacct
     5401 tgtagtcgcc gtagacgaac tcggactcca gcttcgggta cttcttgatc agcgcggtgc
     5461 ccacgacggc gttcaggtac gcgtcgtggg cgtggtggta gttgttgatc tcccggacct
     5521 tgtagaactg gaagtccttg cggaagtcgg agaccagctt gctcttgagg gtgatcacct
     5581 tgacctcgcg gatcagcttg tcgttctcgt cgtacttggt gttcatccgg gagtccagga
     5641 tctgggccac gtgcttggtg atctgccggg tctcgaccag ctggcgcttg atgaagcccg
     5701 ccttgtcgag ctcgctcagg ccgccccgct cggccttggt caggttgtcg aacttgcgct
     5761 gggtgatgag cttggcgttg agcagctggc gccagtagtt cttcatcttc ttcacgacct
     5821 cctccgaggg cacgttgtcg gacttgcccc ggttcttgtc cgagcgggtc aggaccttgt
     5881 tgtcgatcga gtcgtccttc aggaaggact gcggcacgat gtggtccacg tcgtagtcgg
     5941 acagccggtt gatgtcgagc tcctggtcca cgtacatgtc gcggccgttc tgcaggtagt
     6001 agaggtacag cttctcgttc tgcagctggg tgttctcgac cgggtgctcc ttcaggatct
     6061 ggctgccgag ctccttgatg ccctcctcga tccgcttcat ccgctcgcgc gagttcttct
     6121 ggcccttctg ggtggtctgg ttctcccggg ccatctcgat cacgatgttc tcgggcttgt
     6181 ggcggcccat caccttgacc agctcgtcca cgaccttgac ggtctggagg atgcccttct
     6241 tgatcgccgg ggagcccgcc aggttggcga tgtgctcgtg gaggctgtcg ccctggcccg
     6301 agacctgggc cttctggatg tcctccttga aggtcaggct gtcgtcgtgg atgagctgca
     6361 tgaagttgcg gttggcgaag ccgtcggact tgaggaagtc caggatggtc ttgccgctct
     6421 gcttgtcccg gatgccgttg atcagcttcc gggagaggcg gccccagccg gtgtagcggc
     6481 gacgcttcag ctgcttcatc accttgtcgt cgaacaggtg ggcgtaggtc ttgagccgct
     6541 cctcgatcat ctcgcggtcc tcgaagaggg tcagggtgag gacgatgtcc tccaggatgt
     6601 cctcgttctc ctcgttgtcg aggaagtcct tgtccttgat gatcttgagc aggtcgtggt
     6661 aggtgcccag ggaggcgttg aagcggtcct ccacgccgct gatctcgacg gagtcgaagc
     6721 actcgatctt cttgaagtag tcctccttca gctgcttcac ggtgaccttg cggttggtct
     6781 tgaagagcag gtccacgatc gccttcttct gctcgcccga caggaaggcc ggcttccgca
     6841 tgccctcggt cacgtacttg accttggtca gctcgttgta cacggtgaag tactcgtaga
     6901 gcagggagtg cttgggcagg accttctcgt tcgggaggtt cttgtcgaag ttggtcatgc
     6961 gctcgatgaa cgactgcgcg gacgcgccct tgtccacgac ctcctcgaag ttccacgggg
     7021 tgatggtctc ctccgacttc cgggtcatcc acgcgaaccg ggagttgccg cgggccaggg
     7081 ggccgacgta gtacgggatg cggaaggtca ggatcttctc gatcttctcg cggttgtcct
     7141 tcaggaaggg gtagaagtcc tcctggcgcc ggaggatggc gtggagctcg cccaggtgga
     7201 tctggtgcgg gatggagccg ttgtcgaagg tccgctgctt gcggagcagg tcctcgcggt
     7261 tcagcttgac gagcagctcc tcggtgccgt ccatcttctc caggatcggc ttgatgaact
     7321 tgtagaactc ctcctgcgac gcgccgccgt cgatgtagcc ggcgtagccg ttcttggact
     7381 ggtcgaagaa gatttccttg tacttctcgg gcagctgctg gcgcacgagg gccttgagca
     7441 gggtcaggtc ctggtggtgc tcgtcgtacc gcttgatcat gctcgccgac agcggggcct
     7501 tggtgatctc ggtgttgacc cgcaggatgt cgctgagcag gatggcgtcc gagaggttct
     7561 tcgcggccag gaagaggtcc gcgtactggt cgccgatctg ggcgagcagg ttgtccaggt
     7621 cgtcgtcgta ggtgtccttg gacagctgga gcttcgcgtc ctcggccagg tcgaagttgc
     7681 tcttgaagtt gggggtcagg ccgagcgaca gcgcgatcag gttgccgaag aggccgttct
     7741 tcttctcgcc cggcagctgg gcgatgaggt tctccaggcg ccgggacttg ctcaggcgcg
     7801 cggagaggat cgccttggcg tccacgccgc tggcgttgat ggggttctcc tcgaacagct
     7861 ggttgtaggt ctgcaccagc tggatgaaga gcttgtccac gtcggagttg tccgggttca
     7921 ggtcgccctc gatgaggaag tggccgcgga acttgatcat gtgcgcgagg gccaggtaga
     7981 tgagccgcag gtccgccttg tcggtcgagt ccaccagctt cttgcggagg tggtagatgg
     8041 tggggtactt ctcgtggtag gccacctcgt ccacgatgtt gccgaagatc gggtggcgct
     8101 cgtgcttctt gtcctcctcc accaggaagc tctcctccag gcggtggaag aacgagtcgt
     8161 ccaccttggc catctcgttg gagaagattt cctgcaggta gcagatgcgg ttcttgcgcc
     8221 gggtgtagcg gcgacgcgcg gtccgcttca ggcgggtcgc ctcggcggtc tcgccgctgt
     8281 cgaagagcag ggcgccgatc aggttcttct tgatcgagtg ccggtcggtg ttgcccagga
     8341 ccttgaactt cttggagggg accttgtact cgtcggtgat gaccgcccag cccacggagt
     8401 tggtgccgat cgcgaggccg atggagtact tcttgtcgga ctccggggtg gcggactccg
     8461 aggtgcccgg ggtctcggag ccggacttca ggccggtggc ccacaggatg tggggcggca
     8521 ggcgctggta gtggcaggac tggagggcga tggtgaagaa ggtcagctgc ggctgcttgc
     8581 gacgcaggat gttcaggcag ggcggcaggc ccaggatgat gcagtacagc tccaggacgt
     8641 acaggcggac ccacaggtgc gggtagcggg gccagtgggc ctcgttggac ggggagtagt
     8701 tgacgaagtt ccgccagcag tagccggact cctgctcggt catgatctgg atggtgacgc
     8761 cggaggagat caggtcgcgc aggccctgcc ggttgcgcgg gtcggcgtgg tggtacagcc
     8821 gggcgatgta gatgaacagg gtgacgtgcg ggtagcggga caggaactcg gtgatggccc
     8881 gggagcactc gccgcagggg gaccaggaca ggaaccaggt gatggagcag cgggtgttcg
     8941 ggcagaagta ccgctcggtg gtgaacttct cgatgaagtt gacctcgacg tgcttgttgg
     9001 tgttctgcga ggtgtggcgc cagatggagt gccggccgcc ccagttgatc tcgtacagca
     9061 ggcaggtctc cttgcgcagc tcgcgcgggt cgaagaagac ctcgaactcg tgcggctcga
     9121 tgcggcgacg cagggtcggg tccacggcca cggggccggt ctcggaggac atatgtccgc
     9181 tcccttctct gacgccgtcc acgctgcctc ctcacgtgac gtgaggtgca agcccggacg
     9241 ttccgcgtgc cacgccgtga gccgccgcgt gccgtcggct ccctcagccc agatccccga
     9301 tccggtccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     9361 cgggcgtttt ttattggtga gaatcgcagc aacttgtcgc gccaatcgag ccatgtcgtc
     9421 gtcaacgacc ccccattcaa gaacagcaag cagcattgag aactttggaa tccagtccct
     9481 cttccacctg ctgaccggat cagcagtccc cggaacatcg tagctgacgc ctactcagaa
     9541 ccagggaacg gaacctcctt tgctcaaccc gtaacaggtg aatcctcctt cctctgccgt
     9601 cacctgcaac ggcccgtggc ggatgaagag acggaagtgc tgtccggtgc tctggctgcg
     9661 tagcgtgacg aagggcaggt ccaaggctct cgcgaccgta tcgggaatgc gtttccgagc
     9721 ctcctcctca ctcagatcgt gccggcgcat gagccgccgc cgcaggcgtt ccggattgct
     9781 tttcgcctga acccgactga cctgacggta cggtgtgggg tgaggcacga ctgccggttc
     9841 tccgaattgc agatggtccc gcaacccttc cagccagggc cgggcgagca gggcacgaag
     9901 gtcgtccgcc gaggcatgaa tgcgcaggcg ctcgcccagc cgggagcggc tttcgtcgag
     9961 gtcggggaag ctcacgccga tcctgtcccc gccctgtgcc accagggcct ggtggagctt
    10021 gccgaagagc acgctcatga gttgcgccgg gggaaattcc gggtccggtc gcaagcgaat
    10081 gtcgaggtag tggtccatgc cgtcgatcct accaaccggc acgattgtgc ccacaacagc
    10141 atcgcggtgc cacgtgtgga ccgcgtcggt cagatcctcc ccgcacctct cgccagccgt
    10201 caagatcgac cgcgtacaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
    10261 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
    10321 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
    10381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
    10441 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
    10501 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
    10561 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
    10621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
    10681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
    10741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
    10801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
    10861 ggtcgttcgc tccaagctgg gctgtgtgca cgaccccccg ttcagcccga ccgctgcgcc
    10921 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    10981 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    11041 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    11101 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    11161 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    11221 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    11281 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt ggggtgggcg
    11341 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
    11401 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
    11461 ttgggcgtcg cttggtcggt catttcgaat aggaacttcg gaataggaac ttatgagctc
    11521 agccaatcga ctggcgagcg gcatcgcatt cttcgcatcc cgcctctggc ggatgcagga
    11581 agatcaacgg atctcggccc agttgaccca gggctgtcgc cacaatgtcg cgggagcgga
    11641 tcaaccgagc aaaggcatga ccgactggac cttccttctg aaggctcttc tccttgagcc
    11701 acctgtccgc caaggcaaag cgctcacagc agtggtcatt ctcgagataa tcgacgcgta
    11761 ccaacttgcc atcctgaaga atggtgcagt gtctcggcac cccataggga acctttgcca
    11821 acaactcggc aagatgcagc gtcgtgttgg catcgtgtcc cacgccgagg agaagtacct
    11881 gcccatcgag ttcatggaca cgggcgaccg ggcttgcagg cgagtgaggt ggcaggggca
    11941 atggatcaga gatgatctgc tctgcctgtg gccccgctgc cgcaaaggca aatggatggg
    12001 cgctgcgctt tacatttggc aggcgccaga atgagtcaga gacaactcca aggtccggtg
    12061 taacgggcga cgtggcagga tcgaacggct cgtcgtccag acctgaccac gagggcatga
    12121 cgagcgtccc tcccggaccc agcgcagcac gcagggcctc gatcagtcca agtggcccat
    12181 cttcgagggg ccggacgcta cggaaggagc tgtggaccag cagcacaccg ccgggggtaa
    12241 ccccaaggtt gagaagctga ccgatgagct cggcttttcg ccattcgtat tgcacgacat
    12301 tgcactccac cgctgatgac atcagtcgat catagcacga tcaacggcac tgttgcaaat
    12361 agtcggtggt gataaactta tcatcccctt ttgcttatgg agctgcacat gaacccattc
    12421 aaaggccggc attttcagcg tgacatcatt ctgtgggccg tacgctggta ctgcaaatac
    12481 ggcatcagtt accgtgagct gcattttccg ctgcataacc ctgcttcggg gtcattatag
    12541 cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc
    12601 gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc
    12661 cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg
    12721 ggaatcctgc tctgcgaggc tggcgggaac tctagctcaa cgcgaaggcc cgccccggtg
    12781 agtggggcgg accttcgtgc tgtccagggt gccccagccc gggtgccggg tgggcgccat
    12841 ctgcccgtaa tcctccgtgg cctggccgta cggcgtctgc ggaccggccg ggcggtgaaa
    12901 gacggcaggc gggaagggag gggtggtcgc ggtccctcag cggccgtctg acgcccgatc
    12961 acggggctga ccgccgcacc cacgccgtca cccctgcggg cgcgtgaggc gaccatccag
    13021 ccagccgtgg cgggcgggga gctcctgctg ccgcgatccg ctgcggcccg gcacgggtcg
    13081 cgtaccgctt cgggcccagc atcgtcgtgg ccgaggcatc ggtcgaccaa gggaccggtc
    13141 cgacccggcg gacaaccgcg gcccggcccg ccggccaaaa gcaggcaggc ggacgcgtcg
    13201 tcgtcgacgg cc
//
