LOCUS       sequence-367542-       12013 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. Gentamycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12013)
  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 12013)
  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: pMBEC6
FEATURES             Location/Qualifiers
     source          1..12013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..23,24..30)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_RNA        781..856
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        join(891..896,897..913,914..920)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    928..944
                     /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             997..1560
                     /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             1718..2404
                     /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(2453..2476,2477..2479,2480..6553)
                     /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             6566..6817
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        join(6877..6882,6883..6899,6900..6905)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             7094..7627
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      7925..8276
                     /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      9294..9388
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      9476..10064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9965..9984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10155..10174
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(10213..11547,11548..11634))
                     /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"
ORIGIN
        1 ttgacaatta atcatccggc tcgtataatg tgtggcgaga ccaggaggaa aaacatatgc
       61 gtaaatgtga agaactgttc accggtgttg ttccgatcct ggttgaactg gatggtgatg
      121 ttaacggcca caaattctct gttcgtggtg aaggtgaagg tgatgcaacc aacggtaaac
      181 tgaccctgaa attcatctgc actaccggta aactgccggt tccatggccg actctggtga
      241 ctaccctgac ctatggtgtt cagtgttttt ctcgttaccc ggatcacatg aagcagcatg
      301 atttcttcaa atctgcaatg ccggaaggtt atgtacagga gcgcaccatt tctttcaaag
      361 acgatggcac ctacaaaacc cgtgcagagg ttaaatttga aggtgatact ctggtgaacc
      421 gtattgaact gaaaggcatt gatttcaaag aggacggcaa catcctgggc cacaaactgg
      481 aatataactt caactcccat aacgtttaca tcaccgcaga caaacagaag aacggtatca
      541 aagctaactt caaaattcgc cataacgttg aagacggtag cgtacagctg gcggaccact
      601 accagcagaa cactccgatc ggtgatggtc cggttctgct gccggataac cactacctgt
      661 ccacccagtc taaactgtcc aaagacccga acgaaaagcg cgaccacatg gtgctgctgg
      721 agttcgttac tgccgcaggt atcacgcacg gcatggatga actctacaaa taaggtctca
      781 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      841 ggcaccgagt cggtgctttt tttgagatct gtccataccc atggtctaga ttgacaatta
      901 atcatccggc tcgtataatg tgtggaattg tgagcggata acaatttcac accctaggcc
      961 gcggccgcgc gaattcaagc cgaagagaag taagccatgg accactacct cgacattcgc
     1021 ttgcgaccgg acccggaatt tcccccggcg caactcatga gcgtgctctt cggcaagctc
     1081 caccaggccc tggtggcaca gggcggggac aggatcggcg tgagcttccc cgacctcgac
     1141 gaaagccgct cccggctggg cgagcgcctg cgcattcatg cctcggcgga cgaccttcgt
     1201 gccctgctcg cccggccctg gctggaaggg ttgcgggacc atctgcaatt cggagaaccg
     1261 gcagtcgtgc ctcaccccac accgtaccgt caggtcagtc gggttcaggc gaaaagcaat
     1321 ccggaacgcc tgcggcggcg gctcatgcgc cggcacgatc tgagtgagga ggaggctcgg
     1381 aaacgcattc ccgatacggt cgcgagagcc ttggacctgc ccttcgtcac gctacgcagc
     1441 cagagcaccg gacagcactt ccgtctcttc atccgccacg ggccgttgca ggtgacggca
     1501 gaggaaggag gattcacctg ttacgggttg agcaaaggag gtttcgttcc ctggttctga
     1561 ctcgaggcat caacgacaag ccccggctat cccgatagcc ggggctttgt cttgacggtg
     1621 tacaagaacc tctttactat tttgtacccc gaaaattggc agggcgcacg ctctgtcatt
     1681 tcgtttgtgt gtcgtaagac aatcagtgga gctatagatg agtagcgaaa ccggtccggt
     1741 tgcagtggat ccgaccctgc gccgtcgcat tgaaccgcac gagtttgaag tgttctttga
     1801 tccgcgcgag ctgcgcaaag aaacttgcct gctgtacgag attaactggg gtggccgcca
     1861 tagcatctgg cgccatacca gccagaacac caataagcac gtggaagtga attttattga
     1921 aaaatttacc accgagcgct acttctgccc taatacccgc tgcagcatca cctggtttct
     1981 gagctggagc ccgtgcggcg aatgtagtcg cgccattacc gagttcctga gccgctatcc
     2041 gcatgtgacc ctgttcatct acatcgcccg tctgtaccat cacgccgatc cgcgcaatcg
     2101 ccaaggtctg cgtgatctga ttagcagcgg tgtgaccatc cagatcatga ccgaacaaga
     2161 gagcggctat tgctggcgca acttcgtgaa ctattctccg agcaacgaag cccactggcc
     2221 gcgttatccg catctgtggg tgcgcctgta tgtgctggag ctgtactgca tcatcctggg
     2281 cctgccgcct tgcctgaata ttctgcgccg taaacagccg caactgacat tcttcaccat
     2341 cgcactgcag agctgccatt atcagcgcct gccgccgcac attttatggg ccaccggtct
     2401 gaaaagcggt agtgaaactc cgggcacaag cgaaagcgca accccggaaa gtgataagaa
     2461 atactcaata ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga tcactgatga
     2521 atataaggtt ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa
     2581 aaaaaatctt ataggggctc ttttatttga cagtggagag acagcggaag cgactcgtct
     2641 caaacggaca gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga
     2701 gattttttca aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc
     2761 ttttttggtg gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga
     2821 tgaagttgct tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga
     2881 ttctactgat aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt
     2941 tcgtggtcat tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact
     3001 atttatccag ttggtacaaa cctacaatca attatttgaa gaaaacccta ttaacgcaag
     3061 tggagtagat gctaaagcga ttctttctgc acgattgagt aaatcaagac gattagaaaa
     3121 tctcattgct cagctccccg gtgagaagaa aaatggctta tttgggaatc tcattgcttt
     3181 gtcattgggt ttgaccccta attttaaatc aaattttgat ttggcagaag atgctaaatt
     3241 acagctttca aaagatactt acgatgatga tttagataat ttattggcgc aaattggaga
     3301 tcaatatgct gatttgtttt tggcagctaa gaatttatca gatgctattt tactttcaga
     3361 tatcctaaga gtaaatactg aaataactaa ggctccccta tcagcttcaa tgattaaacg
     3421 ctacgatgaa catcatcaag acttgactct tttaaaagct ttagttcgac aacaacttcc
     3481 agaaaagtat aaagaaatct tttttgatca atcaaaaaac ggatatgcag gttatattga
     3541 tgggggagct agccaagaag aattttataa atttatcaaa ccaattttag aaaaaatgga
     3601 tggtactgag gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac
     3661 ctttgacaac ggctctattc cccatcaaat tcacttgggt gagctgcatg ctattttgag
     3721 aagacaagaa gacttttatc catttttaaa agacaatcgt gagaagattg aaaaaatctt
     3781 gacttttcga attccttatt atgttggtcc attggcgcgt ggcaatagtc gttttgcatg
     3841 gatgactcgg aagtctgaag aaacaattac cccatggaat tttgaagaag ttgtcgataa
     3901 aggtgcttca gctcaatcat ttattgaacg catgacaaac tttgataaaa atcttccaaa
     3961 tgaaaaagta ctaccaaaac atagtttgct ttatgagtat tttacggttt ataacgaatt
     4021 gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca
     4081 gaagaaagcc attgttgatt tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt
     4141 aaaagaagat tatttcaaaa aaatagaatg ttttgatagt gttgaaattt caggagttga
     4201 agatagattt aatgcttcat taggtaccta ccatgatttg ctaaaaatta ttaaagataa
     4261 agattttttg gataatgaag aaaatgaaga tatcttagag gatattgttt taacattgac
     4321 cttatttgaa gatagggaga tgattgagga aagacttaaa acatatgctc acctctttga
     4381 tgataaggtg atgaaacagc ttaaacgtcg ccgttatact ggttggggac gtttgtctcg
     4441 aaaattgatt aatggtatta gggataagca atctggcaaa acaatattag attttttgaa
     4501 atcagatggt tttgccaatc gcaattttat gcagctgatc catgatgata gtttgacatt
     4561 taaagaagac attcaaaaag cacaagtgtc tggacaaggc gatagtttac atgaacatat
     4621 tgcaaattta gctggtagcc ctgctattaa aaaaggtatt ttacagactg taaaagttgt
     4681 tgatgaattg gtcaaagtaa tggggcggca taagccagaa aatatcgtta ttgaaatggc
     4741 acgtgaaaat cagacaactc aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat
     4801 cgaagaaggt atcaaagaat taggaagtca gattcttaaa gagcatcctg ttgaaaatac
     4861 tcaattgcaa aatgaaaagc tctatctcta ttatctccaa aatggaagag acatgtatgt
     4921 ggaccaagaa ttagatatta atcgtttaag tgattatgat gtcgatcaca ttgttccaca
     4981 aagtttcctt aaagacgatt caatagacaa taaggtctta acgcgttctg ataaaaatcg
     5041 tggtaaatcg gataacgttc caagtgaaga agtagtcaaa aagatgaaaa actattggag
     5101 acaacttcta aacgccaagt taatcactca acgtaagttt gataatttaa cgaaagctga
     5161 acgtggaggt ttgagtgaac ttgataaagc tggttttatc aaacgccaat tggttgaaac
     5221 tcgccaaatc actaagcatg tggcacaaat tttggatagt cgcatgaata ctaaatacga
     5281 tgaaaatgat aaacttattc gagaggttaa agtgattacc ttaaaatcta aattagtttc
     5341 tgacttccga aaagatttcc aattctataa agtacgtgag attaacaatt accatcatgc
     5401 ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg attaagaaat atccaaaact
     5461 tgaatcggag tttgtctatg gtgattataa agtttatgat gttcgtaaaa tgattgctaa
     5521 gtctgagcaa gaaataggca aagcaaccgc aaaatatttc ttttactcta atatcatgaa
     5581 cttcttcaaa acagaaatta cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga
     5641 aactaatggg gaaactggag aaattgtctg ggataaaggg cgagattttg ccacagtgcg
     5701 caaagtattg tccatgcccc aagtcaatat tgtcaagaaa acagaagtac agacaggcgg
     5761 attctccaag gagtcaattt taccaaaaag aaattcggac aagcttattg ctcgtaaaaa
     5821 agactgggat ccaaaaaaat atggtggttt tgatagtcca acggtagctt attcagtcct
     5881 agtggttgct aaggtggaaa aagggaaatc gaagaagtta aaatccgtta aagagttact
     5941 agggatcaca attatggaaa gaagttcctt tgaaaaaaat ccgattgact ttttagaagc
     6001 taaaggatat aaggaagtta aaaaagactt aatcattaaa ctacctaaat atagtctttt
     6061 tgagttagaa aacggtcgta aacggatgct ggctagtgcc ggagaattac aaaaaggaaa
     6121 tgagctggct ctgccaagca aatatgtgaa ttttttatat ttagctagtc attatgaaaa
     6181 gttgaagggt agtccagaag ataacgaaca aaaacaattg tttgtggagc agcataagca
     6241 ttatttagat gagattattg agcaaatcag tgaattttct aagcgtgtta ttttagcaga
     6301 tgccaattta gataaagttc ttagtgcata taacaaacat agagacaaac caatacgtga
     6361 acaagcagaa aatattattc atttatttac gttgacgaat cttggagctc ccgctgcttt
     6421 taaatatttt gatacaacaa ttgatcgtaa acgatatacg tctacaaaag aagttttaga
     6481 tgccactctt atccatcaat ccatcactgg tctttatgaa acacgcattg atttgagtca
     6541 gctaggaggt gactccggcg gctccaccaa cctgtccgac atcatcgaga aggagactgg
     6601 caagcagctg gtcatccagg agtccatcct gatgctgccg gaggaagtcg aggaagtcat
     6661 cggcaacaag cccgagtccg acatcctggt ccacaccgcc tacgacgagt ccaccgacga
     6721 gaacgtcatg ctgctgacct cggacgcccc cgagtacaag ccctgggccc tggtcatcca
     6781 ggactccaac ggcgagaaca agatcaagat gctgtgaact tcggaatagg aacttcaaga
     6841 tcccctgatt ccctttgtca acagcaatgg atcgaattga cataagcctg ttcggttcgt
     6901 aaactgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
     6961 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttgtacag
     7021 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     7081 tggagcagca acgatgttac gcagcagcaa cgatgttacg cagcagggca gtcgccctaa
     7141 aacaaagtta ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg gccctgacca
     7201 agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcggag acgtagccac
     7261 ctactcccaa catcagccgg actccgatta cctcgggaac ttgctccgta gtaagacatt
     7321 catcgcgctt gctgccttcg accaagaagc ggttgttggc gctctcgcgg cttacgttct
     7381 gcccaagttt gagcagccgc gtagtgagat ctatatctat gatctcgcag tctccggcga
     7441 gcaccggagg cagggcattg ccaccgcgct catcaatctc ctcaagcatg aggccaacgc
     7501 gcttggtgct tatgtgatct acgtgcaagc agattacggt gacgatcccg cagtggctct
     7561 ctatacaaag ttgggcatac gggaagaagt gatgcacttt gatatcgacc caagtaccgc
     7621 cacctaacaa ttcgttcaag ccgagatcag cttcccggcc gcggagttgt tcggtaaatt
     7681 gtcacaacgc cgcggccaat tcgagagttt tcgttccact gagcgtcaga ccccaattac
     7741 acgccactgg ctgtgcttgc tggggtgacg gtggcaacgg tggcggcctt gctgggctat
     7801 cgcgttggaa agaaacgagg gaaaggggac tgataaaccg gtcttagccc ctccccttgg
     7861 tgtccaaccg ctctgtaggc ctctcaggcg ccgctggtgc cgctggttgg acgccaaggg
     7921 tgaatccgcc tcgataccct gattactcgc ttcctgcgcc ctctcaggcg gcgatagggg
     7981 actggtaaaa cggggattgc ccagacgcct cccccgcccc ttcaggggca caaatgcggc
     8041 cccaacgggg ccacgtagtg gtgcgttttt tgcgtttcca cccttttctt ccttttccct
     8101 tttaaacctt ttaggacgtc tacaggccac gtaatccgtg gcctgtagag tttaaaaagg
     8161 gacggatttg ttgccattaa gggacggatt tgttgttaag aagggacgga tttgttgttg
     8221 taaagggacg gatttgttgt attgtgggac gcagatacag tgtcccctta tacacaagga
     8281 atgtcgaacg tggcctcacc cccaatggtt tacaaaagca atgccctggt cgaggccgcg
     8341 tatcgcctca gtgttcagga acagcggatc gttctggcct gtattagcca ggtgaagagg
     8401 agcgagcctg tcaccgatga agtgatgtat tcagtgacgg cggaggacat agcgacgatg
     8461 gcgggtgtcc ctatcgaatc ttcctacaac cagctcaaag aagcggccct gcgcctgaaa
     8521 cggcgggaag tccggttaac ccaagagccc aatggcaagg ggaaaagacc gagtgtgatg
     8581 attaccggct gggtgcaaac aatcatctac cgggagggtg agggccgtgt agaactcagg
     8641 ttcaccaaag acatgctgcc gtacctgacg gaactcacca aacagttcac caaatacgcc
     8701 ttggctgacg tggccaagat ggacagcacc cacgcgatca ggctttacga gctgctcatg
     8761 caatgggaca gcatcggcca gcgcgaaata gaaattgacc agctgcgaaa gtggtttcaa
     8821 ctggaaggcc ggtatccctc gatcaaggac ttcaagttgc gagtgcttga tccagccgtg
     8881 acgcagatca acgagcacag cccgctacag gtggagtggg cgcagcgaaa gaccgggcgc
     8941 aaggtcacac atctgttgtt cagttttgga ccgaagaagc ccgccaaggc ggtgggtaag
     9001 gccccagcga agcgcaaggc cgggaagatt tcagatgctg agatcgcgaa acaggctcgc
     9061 cctggtgaga catgggaagc ggcccgcgct cgactaaccc agatgccgct tgcgcaaact
     9121 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     9181 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     9241 taaatctgga gccggtgagc gtggcgggac tctggggttc gagagctcgc ttggactcct
     9301 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     9361 ccgccgggcg ttttttattg gtgagaatcc aagcactagt catgaccaaa atcccttaac
     9421 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9481 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9541 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9601 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     9661 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     9721 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     9781 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     9841 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     9901 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     9961 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
    10021 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
    10081 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttgt
    10141 cgacagagcc cgggcggcat cagagcagat tgtactgaga gtgcaccata atcggcattt
    10201 tcttttgcgt ttttatttgt taactgttaa ttgtccttgt tcaaggatgc tgtctttgac
    10261 aacagatgtt ttcttgcctt tgatgttcag caggaagcta ggcgcaaacg ttgattgttt
    10321 gtctgcgtag aatcctctgt ttgtcatata gcttgtaatc acgacattgt ttcctttcgc
    10381 ttgaggtaca gcgaagtgtg agtaagtaaa ggttacatcg ttaggatcaa gatccatttt
    10441 taacacaagg ccagttttgt tcagcggctt gtatgggcca gttaaagaat tagaaacata
    10501 accaagcatg taaatatcgt tagacgtaat gccgtcaatc gtcatttttg atccgcggga
    10561 gtcagtgaac agataccatt tgccgttcat tttaaagacg ttcgcgcgtt caatttcatc
    10621 tgttactgtg ttagatgcaa tcagcggttt catcactttt ttcagtgtgt aatcatcgtt
    10681 tagctcaatc ataccgagag cgccgtttgc taactcagcc gtgcgttttt tatcgctttg
    10741 cagaagtttt tgactttctt gacggaagaa tgatgtgctt ttgccatagt atgctttgtt
    10801 aaataaagat tcttcgcctt ggtagccatc ttcagttcca gtgtttgctt caaatactaa
    10861 gtatttgtgg cctttatctt ctacgtagtg aggatctctc agcgtatggt tgtcgcctga
    10921 gctgtagttg ccttcatcga tgaactgctg tacattttga tacgtttttc cgtcaccgtc
    10981 aaagattgat ttataatcct ctacaccgtt gatgttcaaa gagctgtctg atgctgatac
    11041 gttaacttgt gcagttgtca gtgtttgttt gccgtaatgt ttaccggaga aatcagtgta
    11101 gaataaacgg atttttccgt cagatgtaaa tgtggctgaa cctgaccatt cttgtgtttg
    11161 gtcttttagg atagaatcat ttgcatcgaa tttgtcgctg tctttaaaga cgcggccagc
    11221 gtttttccag ctgtcaatag aagtttcgcc gactttttga tagaacatgt aaatcgatgt
    11281 gtcatccgca tttttaggat ctccggctaa tgcaaagacg atgtggtagc cgtgatagtt
    11341 tgcgacagtg ccgtcagcgt tttgtaatgg ccagctgtcc caaacgtcca ggccttttgc
    11401 agaagagata tttttaattg tggacgaatc gaactcagga acttgatatt tttcattttt
    11461 ttgctgttca gggatttgca gcatatcatg gcgtgtaata tgggaaatgc cgtatgtttc
    11521 cttatatggc ttttggttcg tttctttcgc aaacgcttga gttgcgcctc ctgccagcag
    11581 tgcggtagta aaggttaata ctgttgcttg ttttgcaaac tttttgatgt tcatcgttca
    11641 tgtctccttt tttatgtact gtgttagcgg tctgcttctt ccagccctcc tgtttgaaga
    11701 tggcaagtta gttacgcaca ataaaaaaag acctaaaata tgtaaggggt gacgccaaag
    11761 tatacacttt gccctttaca cattttaggt cttgcctgct ttatcagtaa caaacccgcg
    11821 cgatttactt ttcgacctca ttctattaga ctctcgtttg gattgcaact ggtctatttt
    11881 cctcttttgt ttgatagaaa atcataaaag gatttgcaga ctacgggcct aaagaactaa
    11941 aaaatctatc tgtttctttt cattctctgt attttttata gtttctgttg catgggcata
    12001 aagttgcctt aag
//
