LOCUS       sequence-367572-       12242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Base editing plasmid, constitutively expressing cytosine base editor
            and cas6. Contains GFP, flanked by BsaI restriction sites to 
            introduce spacer and gRNAs. Apramycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12242)
  AUTHORS   Volke DC, Martino RA, Kozaeva E, Smania AM, Nikel PI
  TITLE     Modular (de)construction of complex bacterial phenotypes by 
            CRISPR/nCas9-assisted, multiplex cytidine base-editing.
  JOURNAL   Nat Commun. 2022 May 31;13(1):3026. doi: 10.1038/s41467-022-30780-z.
  PUBMED    35641501
REFERENCE   2  (bases 1 to 12242)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-05-31"; volume: "13(1):3026. doi"; pages: " 
            10.1038/s41467-022-30780-z"
COMMENT     Sequence Label: pMBEC8
FEATURES             Location/Qualifiers
     source          1..12242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2506..2757
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             2993..3793
                     /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      4094..4445
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     terminator      5463..5557
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5645..6233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6134..6153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6324..6343
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(6382..7716,7717..7803))
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        join(8183..8188,8189..8205,8206..8212)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_RNA        8963..9038
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        join(9073..9078,9079..9095,9096..9102)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    9110..9126
                     /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)."
     CDS             9179..9742
                     /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
                     LQVTAEEGGFTCYGLSKGGFVPWF"
     CDS             9900..10586
                     /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(10635..10658,10659..10661,10662..12242,1..2493)
                     /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"
ORIGIN
        1 gcatttcttt caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga
       61 aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt
      121 gttgaaattt caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg
      181 ctaaaaatta ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag
      241 gatattgttt taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa
      301 acatatgctc acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact
      361 ggttggggac gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa
      421 acaatattag attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc
      481 catgatgata gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc
      541 gatagtttac atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt
      601 ttacagactg taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa
      661 aatatcgtta ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg
      721 cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa
      781 gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa
      841 aatggaagag acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat
      901 gtcgatcaca ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta
      961 acgcgttctg ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa
     1021 aagatgaaaa actattggag acaacttcta aacgccaagt taatcactca acgtaagttt
     1081 gataatttaa cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc
     1141 aaacgccaat tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt
     1201 cgcatgaata ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc
     1261 ttaaaatcta aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag
     1321 attaacaatt accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg
     1381 attaagaaat atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat
     1441 gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc
     1501 ttttactcta atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt
     1561 cgcaaacgcc ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg
     1621 cgagattttg ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa
     1681 acagaagtac agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac
     1741 aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca
     1801 acggtagctt attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta
     1861 aaatccgtta aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat
     1921 ccgattgact ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa
     1981 ctacctaaat atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc
     2041 ggagaattac aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat
     2101 ttagctagtc attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg
     2161 tttgtggagc agcataagca ttatttagat gagattattg agcaaatcag tgaattttct
     2221 aagcgtgtta ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat
     2281 agagacaaac caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat
     2341 cttggagctc ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg
     2401 tctacaaaag aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa
     2461 acacgcattg atttgagtca gctaggaggt gactccggcg gctccaccaa cctgtccgac
     2521 atcatcgaga aggagactgg caagcagctg gtcatccagg agtccatcct gatgctgccg
     2581 gaggaagtcg aggaagtcat cggcaacaag cccgagtccg acatcctggt ccacaccgcc
     2641 tacgacgagt ccaccgacga gaacgtcatg ctgctgacct cggacgcccc cgagtacaag
     2701 ccctgggccc tggtcatcca ggactccaac ggcgagaaca agatcaagat gctgtgaact
     2761 tcggaatagg aacttcaaga tcccctgatt ccctttgtca acagcaatgg atcgaactca
     2821 cggtaactga tgccgtattt gcagtaccag cgtacggccc acagaatgat gtcacgctga
     2881 aaatgccggc ctttgaatgg gttcatgtgc agctccataa gcaaaagggg atgataagtt
     2941 tatcaccacc gactatttgc aacagtgccg ttgatcgtgc tatgatcgac tgatgtcatc
     3001 agcggtggag tgcaatgtcg tgcaatacga atggcgaaaa gccgagctca tcggtcagct
     3061 tctcaacctt ggggttaccc ccggcggtgt gctgctggtc cacagctcct tccgtagcgt
     3121 ccggcccctc gaagatgggc cacttggact gatcgaggcc ctgcgtgctg cgctgggtcc
     3181 gggagggacg ctcgtcatgc cctcgtggtc aggtctggac gacgagccgt tcgatcctgc
     3241 cacgtcgccc gttacaccgg accttggagt tgtctctgac tcattctggc gcctgccaaa
     3301 tgtaaagcgc agcgcccatc catttgcctt tgcggcagcg gggccacagg cagagcagat
     3361 catctctgat ccattgcccc tgccacctca ctcgcctgca agcccggtcg cccgtgtcca
     3421 tgaactcgat gggcaggtac ttctcctcgg cgtgggacac gatgccaaca cgacgctgca
     3481 tcttgccgag ttgttggcaa aggttcccta tggggtgccg agacactgca ccattcttca
     3541 ggatggcaag ttggtacgcg tcgattatct cgagaatgac cactgctgtg agcgctttgc
     3601 cttggcggac aggtggctca aggagaagag ccttcagaag gaaggtccag tcggtcatgc
     3661 ctttgctcgg ttgatccgct cccgcgacat tgtggcgaca gccctgggtc aactgggccg
     3721 agatccgttg atcttcctgc atccgccaga ggcgggatgc gaagaatgcg atgccgctcg
     3781 ccagtcgatt ggctgacaat tcgttcaagc cgagatcagc ttcccggccg cggagttgtt
     3841 cggtaaattg tcacaacgcc gcggccaatt cgagagtttt cgttccactg agcgtcagac
     3901 cccaattaca cgccactggc tgtgcttgct ggggtgacgg tggcaacggt ggcggccttg
     3961 ctgggctatc gcgttggaaa gaaacgaggg aaaggggact gataaaccgg tcttagcccc
     4021 tccccttggt gtccaaccgc tctgtaggcc tctcaggcgc cgctggtgcc gctggttgga
     4081 cgccaagggt gaatccgcct cgataccctg attactcgct tcctgcgccc tctcaggcgg
     4141 cgatagggga ctggtaaaac ggggattgcc cagacgcctc ccccgcccct tcaggggcac
     4201 aaatgcggcc ccaacggggc cacgtagtgg tgcgtttttt gcgtttccac ccttttcttc
     4261 cttttccctt ttaaaccttt taggacgtct acaggccacg taatccgtgg cctgtagagt
     4321 ttaaaaaggg acggatttgt tgccattaag ggacggattt gttgttaaga agggacggat
     4381 ttgttgttgt aaagggacgg atttgttgta ttgtgggacg cagatacagt gtccccttat
     4441 acacaaggaa tgtcgaacgt ggcctcaccc ccaatggttt acaaaagcaa tgccctggtc
     4501 gaggccgcgt atcgcctcag tgttcaggaa cagcggatcg ttctggcctg tattagccag
     4561 gtgaagagga gcgagcctgt caccgatgaa gtgatgtatt cagtgacggc ggaggacata
     4621 gcgacgatgg cgggtgtccc tatcgaatct tcctacaacc agctcaaaga agcggccctg
     4681 cgcctgaaac ggcgggaagt ccggttaacc caagagccca atggcaaggg gaaaagaccg
     4741 agtgtgatga ttaccggctg ggtgcaaaca atcatctacc gggagggtga gggccgtgta
     4801 gaactcaggt tcaccaaaga catgctgccg tacctgacgg aactcaccaa acagttcacc
     4861 aaatacgcct tggctgacgt ggccaagatg gacagcaccc acgcgatcag gctttacgag
     4921 ctgctcatgc aatgggacag catcggccag cgcgaaatag aaattgacca gctgcgaaag
     4981 tggtttcaac tggaaggccg gtatccctcg atcaaggact tcaagttgcg agtgcttgat
     5041 ccagccgtga cgcagatcaa cgagcacagc ccgctacagg tggagtgggc gcagcgaaag
     5101 accgggcgca aggtcacaca tctgttgttc agttttggac cgaagaagcc cgccaaggcg
     5161 gtgggtaagg ccccagcgaa gcgcaaggcc gggaagattt cagatgctga gatcgcgaaa
     5221 caggctcgcc ctggtgagac atgggaagcg gcccgcgctc gactaaccca gatgccgctt
     5281 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5341 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5401 tattgctgat aaatctggag ccggtgagcg tggcgggact ctggggttcg agagctcgct
     5461 tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     5521 gctcggttgc cgccgggcgt tttttattgg tgagaatcca agcactagtc atgaccaaaa
     5581 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5641 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5701 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5761 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5821 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5881 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5941 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6001 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6061 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6301 ctgcgttgtc gacagagccc gggcggcatc agagcagatt gtactgagag tgcaccataa
     6361 tcggcatttt cttttgcgtt tttatttgtt aactgttaat tgtccttgtt caaggatgct
     6421 gtctttgaca acagatgttt tcttgccttt gatgttcagc aggaagctag gcgcaaacgt
     6481 tgattgtttg tctgcgtaga atcctctgtt tgtcatatag cttgtaatca cgacattgtt
     6541 tcctttcgct tgaggtacag cgaagtgtga gtaagtaaag gttacatcgt taggatcaag
     6601 atccattttt aacacaaggc cagttttgtt cagcggcttg tatgggccag ttaaagaatt
     6661 agaaacataa ccaagcatgt aaatatcgtt agacgtaatg ccgtcaatcg tcatttttga
     6721 tccgcgggag tcagtgaaca gataccattt gccgttcatt ttaaagacgt tcgcgcgttc
     6781 aatttcatct gttactgtgt tagatgcaat cagcggtttc atcacttttt tcagtgtgta
     6841 atcatcgttt agctcaatca taccgagagc gccgtttgct aactcagccg tgcgtttttt
     6901 atcgctttgc agaagttttt gactttcttg acggaagaat gatgtgcttt tgccatagta
     6961 tgctttgtta aataaagatt cttcgccttg gtagccatct tcagttccag tgtttgcttc
     7021 aaatactaag tatttgtggc ctttatcttc tacgtagtga ggatctctca gcgtatggtt
     7081 gtcgcctgag ctgtagttgc cttcatcgat gaactgctgt acattttgat acgtttttcc
     7141 gtcaccgtca aagattgatt tataatcctc tacaccgttg atgttcaaag agctgtctga
     7201 tgctgatacg ttaacttgtg cagttgtcag tgtttgtttg ccgtaatgtt taccggagaa
     7261 atcagtgtag aataaacgga tttttccgtc agatgtaaat gtggctgaac ctgaccattc
     7321 ttgtgtttgg tcttttagga tagaatcatt tgcatcgaat ttgtcgctgt ctttaaagac
     7381 gcggccagcg tttttccagc tgtcaataga agtttcgccg actttttgat agaacatgta
     7441 aatcgatgtg tcatccgcat ttttaggatc tccggctaat gcaaagacga tgtggtagcc
     7501 gtgatagttt gcgacagtgc cgtcagcgtt ttgtaatggc cagctgtccc aaacgtccag
     7561 gccttttgca gaagagatat ttttaattgt ggacgaatcg aactcaggaa cttgatattt
     7621 ttcatttttt tgctgttcag ggatttgcag catatcatgg cgtgtaatat gggaaatgcc
     7681 gtatgtttcc ttatatggct tttggttcgt ttctttcgca aacgcttgag ttgcgcctcc
     7741 tgccagcagt gcggtagtaa aggttaatac tgttgcttgt tttgcaaact ttttgatgtt
     7801 catcgttcat gtctcctttt ttatgtactg tgttagcggt ctgcttcttc cagccctcct
     7861 gtttgaagat ggcaagttag ttacgcacaa taaaaaaaga cctaaaatat gtaaggggtg
     7921 acgccaaagt atacactttg ccctttacac attttaggtc ttgcctgctt tatcagtaac
     7981 aaacccgcgc gatttacttt tcgacctcat tctattagac tctcgtttgg attgcaactg
     8041 gtctattttc ctcttttgtt tgatagaaaa tcataaaagg atttgcagac tacgggccta
     8101 aagaactaaa aaatctatct gtttcttttc attctctgta ttttttatag tttctgttgc
     8161 atgggcataa agttgcctta agttgacaat taatcatccg gctcgtataa tgtgtggcga
     8221 gaccaggagg aaaaacatat gcgtaaatgt gaagaactgt tcaccggtgt tgttccgatc
     8281 ctggttgaac tggatggtga tgttaacggc cacaaattct ctgttcgtgg tgaaggtgaa
     8341 ggtgatgcaa ccaacggtaa actgaccctg aaattcatct gcactaccgg taaactgccg
     8401 gttccatggc cgactctggt gactaccctg acctatggtg ttcagtgttt ttctcgttac
     8461 ccggatcaca tgaagcagca tgatttcttc aaatctgcaa tgccggaagg ttatgtacag
     8521 gagcgcacca tttctttcaa agacgatggc acctacaaaa cccgtgcaga ggttaaattt
     8581 gaaggtgata ctctggtgaa ccgtattgaa ctgaaaggca ttgatttcaa agaggacggc
     8641 aacatcctgg gccacaaact ggaatataac ttcaactccc ataacgttta catcaccgca
     8701 gacaaacaga agaacggtat caaagctaac ttcaaaattc gccataacgt tgaagacggt
     8761 agcgtacagc tggcggacca ctaccagcag aacactccga tcggtgatgg tccggttctg
     8821 ctgccggata accactacct gtccacccag tctaaactgt ccaaagaccc gaacgaaaag
     8881 cgcgaccaca tggtgctgct ggagttcgtt actgccgcag gtatcacgca cggcatggat
     8941 gaactctaca aataaggtct cagttttaga gctagaaata gcaagttaaa ataaggctag
     9001 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttgagat ctgtccatac
     9061 ccatggtcta gattgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
     9121 taacaatttc acaccctagg ccgcggccgc gcgaattcaa gccgaagaga agtaagccat
     9181 ggaccactac ctcgacattc gcttgcgacc ggacccggaa tttcccccgg cgcaactcat
     9241 gagcgtgctc ttcggcaagc tccaccaggc cctggtggca cagggcgggg acaggatcgg
     9301 cgtgagcttc cccgacctcg acgaaagccg ctcccggctg ggcgagcgcc tgcgcattca
     9361 tgcctcggcg gacgaccttc gtgccctgct cgcccggccc tggctggaag ggttgcggga
     9421 ccatctgcaa ttcggagaac cggcagtcgt gcctcacccc acaccgtacc gtcaggtcag
     9481 tcgggttcag gcgaaaagca atccggaacg cctgcggcgg cggctcatgc gccggcacga
     9541 tctgagtgag gaggaggctc ggaaacgcat tcccgatacg gtcgcgagag ccttggacct
     9601 gcccttcgtc acgctacgca gccagagcac cggacagcac ttccgtctct tcatccgcca
     9661 cgggccgttg caggtgacgg cagaggaagg aggattcacc tgttacgggt tgagcaaagg
     9721 aggtttcgtt ccctggttct gactcgaggc atcaacgaca agccccggct atcccgatag
     9781 ccggggcttt gtcttgacgg tgtacaagaa cctctttact attttgtacc ccgaaaattg
     9841 gcagggcgca cgctctgtca tttcgtttgt gtgtcgtaag acaatcagtg gagctataga
     9901 tgagtagcga aaccggtccg gttgcagtgg atccgaccct gcgccgtcgc attgaaccgc
     9961 acgagtttga agtgttcttt gatccgcgcg agctgcgcaa agaaacttgc ctgctgtacg
    10021 agattaactg gggtggccgc catagcatct ggcgccatac cagccagaac accaataagc
    10081 acgtggaagt gaattttatt gaaaaattta ccaccgagcg ctacttctgc cctaataccc
    10141 gctgcagcat cacctggttt ctgagctgga gcccgtgcgg cgaatgtagt cgcgccatta
    10201 ccgagttcct gagccgctat ccgcatgtga ccctgttcat ctacatcgcc cgtctgtacc
    10261 atcacgccga tccgcgcaat cgccaaggtc tgcgtgatct gattagcagc ggtgtgacca
    10321 tccagatcat gaccgaacaa gagagcggct attgctggcg caacttcgtg aactattctc
    10381 cgagcaacga agcccactgg ccgcgttatc cgcatctgtg ggtgcgcctg tatgtgctgg
    10441 agctgtactg catcatcctg ggcctgccgc cttgcctgaa tattctgcgc cgtaaacagc
    10501 cgcaactgac attcttcacc atcgcactgc agagctgcca ttatcagcgc ctgccgccgc
    10561 acattttatg ggccaccggt ctgaaaagcg gtagtgaaac tccgggcaca agcgaaagcg
    10621 caaccacgga aagtgataag aaatactcaa taggcttagc tatcggcaca aatagcgtcg
    10681 gatgggcggt gatcactgat gaatataagg ttccgtctaa aaagttcaag gttctgggaa
    10741 atacagaccg ccacagtatc aaaaaaaatc ttataggggc tcttttattt gacagtggag
    10801 agacagcgga agcgactcgt ctcaaacgga cagctcgtag aaggtataca cgtcggaaga
    10861 atcgtatttg ttatctacag gagatttttt caaatgagat ggcgaaagta gatgatagtt
    10921 tctttcatcg acttgaagag tcttttttgg tggaagaaga caagaagcat gaacgtcatc
    10981 ctatttttgg aaatatagta gatgaagttg cttatcatga gaaatatcca actatctatc
    11041 atctgcgaaa aaaattggta gattctactg ataaagcgga tttgcgctta atctatttgg
    11101 ccttagcgca tatgattaag tttcgtggtc attttttgat tgagggagat ttaaatcctg
    11161 ataatagtga tgtggacaaa ctatttatcc agttggtaca aacctacaat caattatttg
    11221 aagaaaaccc tattaacgca agtggagtag atgctaaagc gattctttct gcacgattga
    11281 gtaaatcaag acgattagaa aatctcattg ctcagctccc cggtgagaag aaaaatggct
    11341 tatttgggaa tctcattgct ttgtcattgg gtttgacccc taattttaaa tcaaattttg
    11401 atttggcaga agatgctaaa ttacagcttt caaaagatac ttacgatgat gatttagata
    11461 atttattggc gcaaattgga gatcaatatg ctgatttgtt tttggcagct aagaatttat
    11521 cagatgctat tttactttca gatatcctaa gagtaaatac tgaaataact aaggctcccc
    11581 tatcagcttc aatgattaaa cgctacgatg aacatcatca agacttgact cttttaaaag
    11641 ctttagttcg acaacaactt ccagaaaagt ataaagaaat cttttttgat caatcaaaaa
    11701 acggatatgc aggttatatt gatgggggag ctagccaaga agaattttat aaatttatca
    11761 aaccaatttt agaaaaaatg gatggtactg aggaattatt ggtgaaacta aatcgtgaag
    11821 atttgctgcg caagcaacgg acctttgaca acggctctat tccccatcaa attcacttgg
    11881 gtgagctgca tgctattttg agaagacaag aagactttta tccattttta aaagacaatc
    11941 gtgagaagat tgaaaaaatc ttgacttttc gaattcctta ttatgttggt ccattggcgc
    12001 gtggcaatag tcgttttgca tggatgactc ggaagtctga agaaacaatt accccatgga
    12061 attttgaaga agttgtcgat aaaggtgctt cagctcaatc atttattgaa cgcatgacaa
    12121 actttgataa aaatcttcca aatgaaaaag tactaccaaa acatagtttg ctttatgagt
    12181 attttacggt ttataacgaa ttgacaaagg tcaaatatgt tactgaagga atgcgaaaac
    12241 ca
//
