LOCUS       sequence-367535-       12235 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. Kanamycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12235)
  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 12235)
  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: pMBEC2
FEATURES             Location/Qualifiers
     source          1..12235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2510..2761
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     primer_bind     complement(2940..2959)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2951..3766
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      4091..4442
                     /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      5460..5554
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5642..6230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6131..6150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6321..6340
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(6379..7713,7714..7800))
                     /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(8180..8185,8186..8202,8203..8209)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_RNA        8960..9035
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        join(9070..9075,9076..9092,9093..9099)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    9107..9123
                     /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             9176..9739
                     /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             9897..10583
                     /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(10632..10655,10656..10658,10659..12235,1..2497)
                     /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 accagcattt ctttcaggtg aacagaagaa agccattgtt gatttactct tcaaaacaaa
       61 tcgaaaagta accgttaagc aattaaaaga agattatttc aaaaaaatag aatgttttga
      121 tagtgttgaa atttcaggag ttgaagatag atttaatgct tcattaggta cctaccatga
      181 tttgctaaaa attattaaag ataaagattt tttggataat gaagaaaatg aagatatctt
      241 agaggatatt gttttaacat tgaccttatt tgaagatagg gagatgattg aggaaagact
      301 taaaacatat gctcacctct ttgatgataa ggtgatgaaa cagcttaaac gtcgccgtta
      361 tactggttgg ggacgtttgt ctcgaaaatt gattaatggt attagggata agcaatctgg
      421 caaaacaata ttagattttt tgaaatcaga tggttttgcc aatcgcaatt ttatgcagct
      481 gatccatgat gatagtttga catttaaaga agacattcaa aaagcacaag tgtctggaca
      541 aggcgatagt ttacatgaac atattgcaaa tttagctggt agccctgcta ttaaaaaagg
      601 tattttacag actgtaaaag ttgttgatga attggtcaaa gtaatggggc ggcataagcc
      661 agaaaatatc gttattgaaa tggcacgtga aaatcagaca actcaaaagg gccagaaaaa
      721 ttcgcgagag cgtatgaaac gaatcgaaga aggtatcaaa gaattaggaa gtcagattct
      781 taaagagcat cctgttgaaa atactcaatt gcaaaatgaa aagctctatc tctattatct
      841 ccaaaatgga agagacatgt atgtggacca agaattagat attaatcgtt taagtgatta
      901 tgatgtcgat cacattgttc cacaaagttt ccttaaagac gattcaatag acaataaggt
      961 cttaacgcgt tctgataaaa atcgtggtaa atcggataac gttccaagtg aagaagtagt
     1021 caaaaagatg aaaaactatt ggagacaact tctaaacgcc aagttaatca ctcaacgtaa
     1081 gtttgataat ttaacgaaag ctgaacgtgg aggtttgagt gaacttgata aagctggttt
     1141 tatcaaacgc caattggttg aaactcgcca aatcactaag catgtggcac aaattttgga
     1201 tagtcgcatg aatactaaat acgatgaaaa tgataaactt attcgagagg ttaaagtgat
     1261 taccttaaaa tctaaattag tttctgactt ccgaaaagat ttccaattct ataaagtacg
     1321 tgagattaac aattaccatc atgcccatga tgcgtatcta aatgccgtcg ttggaactgc
     1381 tttgattaag aaatatccaa aacttgaatc ggagtttgtc tatggtgatt ataaagttta
     1441 tgatgttcgt aaaatgattg ctaagtctga gcaagaaata ggcaaagcaa ccgcaaaata
     1501 tttcttttac tctaatatca tgaacttctt caaaacagaa attacacttg caaatggaga
     1561 gattcgcaaa cgccctctaa tcgaaactaa tggggaaact ggagaaattg tctgggataa
     1621 agggcgagat tttgccacag tgcgcaaagt attgtccatg ccccaagtca atattgtcaa
     1681 gaaaacagaa gtacagacag gcggattctc caaggagtca attttaccaa aaagaaattc
     1741 ggacaagctt attgctcgta aaaaagactg ggatccaaaa aaatatggtg gttttgatag
     1801 tccaacggta gcttattcag tcctagtggt tgctaaggtg gaaaaaggga aatcgaagaa
     1861 gttaaaatcc gttaaagagt tactagggat cacaattatg gaaagaagtt cctttgaaaa
     1921 aaatccgatt gactttttag aagctaaagg atataaggaa gttaaaaaag acttaatcat
     1981 taaactacct aaatatagtc tttttgagtt agaaaacggt cgtaaacgga tgctggctag
     2041 tgccggagaa ttacaaaaag gaaatgagct ggctctgcca agcaaatatg tgaatttttt
     2101 atatttagct agtcattatg aaaagttgaa gggtagtcca gaagataacg aacaaaaaca
     2161 attgtttgtg gagcagcata agcattattt agatgagatt attgagcaaa tcagtgaatt
     2221 ttctaagcgt gttattttag cagatgccaa tttagataaa gttcttagtg catataacaa
     2281 acatagagac aaaccaatac gtgaacaagc agaaaatatt attcatttat ttacgttgac
     2341 gaatcttgga gctcccgctg cttttaaata ttttgataca acaattgatc gtaaacgata
     2401 tacgtctaca aaagaagttt tagatgccac tcttatccat caatccatca ctggtcttta
     2461 tgaaacacgc attgatttga gtcagctagg aggtgactcc ggcggctcca ccaacctgtc
     2521 cgacatcatc gagaaggaga ctggcaagca gctggtcatc caggagtcca tcctgatgct
     2581 gccggaggaa gtcgaggaag tcatcggcaa caagcccgag tccgacatcc tggtccacac
     2641 cgcctacgac gagtccaccg acgagaacgt catgctgctg acctcggacg cccccgagta
     2701 caagccctgg gccctggtca tccaggactc caacggcgag aacaagatca agatgctgtg
     2761 aacttcggaa taggaacttc aagatcccct gattcccttt gtcaacagca atggatcgaa
     2821 ctggggtccc caataattac gatttaaatt tgtgtctcaa aatctctgat gttacattgc
     2881 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2941 aaggggtgtt atgagccata ttcagcgtga aacgagctgt agccgtccgc gtctgaacag
     3001 caacatggat gcggatctgt atggctataa atgggcgcgt gataacgtgg gtcagagcgg
     3061 cgcgaccatt tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc tgaaacatgg
     3121 caaaggcagc gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact ggctgaccga
     3181 atttatgccg ctgccgacca ttaaacattt tattcgcacc ccggatgatg cgtggctgct
     3241 gaccaccgcg attccgggca aaaccgcgtt tcaggtgctg gaagaatatc cggatagcgg
     3301 cgaaaacatt gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca ttccggtgtg
     3361 caactgcccg tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga gccgtatgaa
     3421 caacggcctg gtggatgcga gcgattttga tgatgaacgt aacggctggc cggtggaaca
     3481 ggtgtggaaa gaaatgcata aactgctgcc gtttagcccg gatagcgtgg tgacccacgg
     3541 cgattttagc ctggataacc tgattttcga tgaaggcaaa ctgattggct gcattgatgt
     3601 gggccgtgtg ggcattgcgg atcgttatca ggatctggcc attctgtgga actgcctggg
     3661 cgaatttagc ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg ataatccgga
     3721 tatgaacaaa ctgcaatttc atctgatgct ggatgaattt ttctaataat taattggacc
     3781 gcggtccgcg cgtcaattcg ttcaagccga gatcagcttc ccggccgcgg agttgttcgg
     3841 taaattgtca caacgccgcg gccaattcga gagttttcgt tccactgagc gtcagacccc
     3901 aattacacgc cactggctgt gcttgctggg gtgacggtgg caacggtggc ggccttgctg
     3961 ggctatcgcg ttggaaagaa acgagggaaa ggggactgat aaaccggtct tagcccctcc
     4021 ccttggtgtc caaccgctct gtaggcctct caggcgccgc tggtgccgct ggttggacgc
     4081 caagggtgaa tccgcctcga taccctgatt actcgcttcc tgcgccctct caggcggcga
     4141 taggggactg gtaaaacggg gattgcccag acgcctcccc cgccccttca ggggcacaaa
     4201 tgcggcccca acggggccac gtagtggtgc gttttttgcg tttccaccct tttcttcctt
     4261 ttccctttta aaccttttag gacgtctaca ggccacgtaa tccgtggcct gtagagttta
     4321 aaaagggacg gatttgttgc cattaaggga cggatttgtt gttaagaagg gacggatttg
     4381 ttgttgtaaa gggacggatt tgttgtattg tgggacgcag atacagtgtc cccttataca
     4441 caaggaatgt cgaacgtggc ctcaccccca atggtttaca aaagcaatgc cctggtcgag
     4501 gccgcgtatc gcctcagtgt tcaggaacag cggatcgttc tggcctgtat tagccaggtg
     4561 aagaggagcg agcctgtcac cgatgaagtg atgtattcag tgacggcgga ggacatagcg
     4621 acgatggcgg gtgtccctat cgaatcttcc tacaaccagc tcaaagaagc ggccctgcgc
     4681 ctgaaacggc gggaagtccg gttaacccaa gagcccaatg gcaaggggaa aagaccgagt
     4741 gtgatgatta ccggctgggt gcaaacaatc atctaccggg agggtgaggg ccgtgtagaa
     4801 ctcaggttca ccaaagacat gctgccgtac ctgacggaac tcaccaaaca gttcaccaaa
     4861 tacgccttgg ctgacgtggc caagatggac agcacccacg cgatcaggct ttacgagctg
     4921 ctcatgcaat gggacagcat cggccagcgc gaaatagaaa ttgaccagct gcgaaagtgg
     4981 tttcaactgg aaggccggta tccctcgatc aaggacttca agttgcgagt gcttgatcca
     5041 gccgtgacgc agatcaacga gcacagcccg ctacaggtgg agtgggcgca gcgaaagacc
     5101 gggcgcaagg tcacacatct gttgttcagt tttggaccga agaagcccgc caaggcggtg
     5161 ggtaaggccc cagcgaagcg caaggccggg aagatttcag atgctgagat cgcgaaacag
     5221 gctcgccctg gtgagacatg ggaagcggcc cgcgctcgac taacccagat gccgcttgcg
     5281 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5341 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5401 tgctgataaa tctggagccg gtgagcgtgg cgggactctg gggttcgaga gctcgcttgg
     5461 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     5521 cggttgccgc cgggcgtttt ttattggtga gaatccaagc actagtcatg accaaaatcc
     5581 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5641 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5701 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5761 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5821 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5881 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5941 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6001 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6061 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6121 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6181 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6241 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6301 cgttgtcgac agagcccggg cggcatcaga gcagattgta ctgagagtgc accataatcg
     6361 gcattttctt ttgcgttttt atttgttaac tgttaattgt ccttgttcaa ggatgctgtc
     6421 tttgacaaca gatgttttct tgcctttgat gttcagcagg aagctaggcg caaacgttga
     6481 ttgtttgtct gcgtagaatc ctctgtttgt catatagctt gtaatcacga cattgtttcc
     6541 tttcgcttga ggtacagcga agtgtgagta agtaaaggtt acatcgttag gatcaagatc
     6601 catttttaac acaaggccag ttttgttcag cggcttgtat gggccagtta aagaattaga
     6661 aacataacca agcatgtaaa tatcgttaga cgtaatgccg tcaatcgtca tttttgatcc
     6721 gcgggagtca gtgaacagat accatttgcc gttcatttta aagacgttcg cgcgttcaat
     6781 ttcatctgtt actgtgttag atgcaatcag cggtttcatc acttttttca gtgtgtaatc
     6841 atcgtttagc tcaatcatac cgagagcgcc gtttgctaac tcagccgtgc gttttttatc
     6901 gctttgcaga agtttttgac tttcttgacg gaagaatgat gtgcttttgc catagtatgc
     6961 tttgttaaat aaagattctt cgccttggta gccatcttca gttccagtgt ttgcttcaaa
     7021 tactaagtat ttgtggcctt tatcttctac gtagtgagga tctctcagcg tatggttgtc
     7081 gcctgagctg tagttgcctt catcgatgaa ctgctgtaca ttttgatacg tttttccgtc
     7141 accgtcaaag attgatttat aatcctctac accgttgatg ttcaaagagc tgtctgatgc
     7201 tgatacgtta acttgtgcag ttgtcagtgt ttgtttgccg taatgtttac cggagaaatc
     7261 agtgtagaat aaacggattt ttccgtcaga tgtaaatgtg gctgaacctg accattcttg
     7321 tgtttggtct tttaggatag aatcatttgc atcgaatttg tcgctgtctt taaagacgcg
     7381 gccagcgttt ttccagctgt caatagaagt ttcgccgact ttttgataga acatgtaaat
     7441 cgatgtgtca tccgcatttt taggatctcc ggctaatgca aagacgatgt ggtagccgtg
     7501 atagtttgcg acagtgccgt cagcgttttg taatggccag ctgtcccaaa cgtccaggcc
     7561 ttttgcagaa gagatatttt taattgtgga cgaatcgaac tcaggaactt gatatttttc
     7621 atttttttgc tgttcaggga tttgcagcat atcatggcgt gtaatatggg aaatgccgta
     7681 tgtttcctta tatggctttt ggttcgtttc tttcgcaaac gcttgagttg cgcctcctgc
     7741 cagcagtgcg gtagtaaagg ttaatactgt tgcttgtttt gcaaactttt tgatgttcat
     7801 cgttcatgtc tcctttttta tgtactgtgt tagcggtctg cttcttccag ccctcctgtt
     7861 tgaagatggc aagttagtta cgcacaataa aaaaagacct aaaatatgta aggggtgacg
     7921 ccaaagtata cactttgccc tttacacatt ttaggtcttg cctgctttat cagtaacaaa
     7981 cccgcgcgat ttacttttcg acctcattct attagactct cgtttggatt gcaactggtc
     8041 tattttcctc ttttgtttga tagaaaatca taaaaggatt tgcagactac gggcctaaag
     8101 aactaaaaaa tctatctgtt tcttttcatt ctctgtattt tttatagttt ctgttgcatg
     8161 ggcataaagt tgccttaagt tgacaattaa tcatccggct cgtataatgt gtggcgagac
     8221 caggaggaaa aacatatgcg taaatgtgaa gaactgttca ccggtgttgt tccgatcctg
     8281 gttgaactgg atggtgatgt taacggccac aaattctctg ttcgtggtga aggtgaaggt
     8341 gatgcaacca acggtaaact gaccctgaaa ttcatctgca ctaccggtaa actgccggtt
     8401 ccatggccga ctctggtgac taccctgacc tatggtgttc agtgtttttc tcgttacccg
     8461 gatcacatga agcagcatga tttcttcaaa tctgcaatgc cggaaggtta tgtacaggag
     8521 cgcaccattt ctttcaaaga cgatggcacc tacaaaaccc gtgcagaggt taaatttgaa
     8581 ggtgatactc tggtgaaccg tattgaactg aaaggcattg atttcaaaga ggacggcaac
     8641 atcctgggcc acaaactgga atataacttc aactcccata acgtttacat caccgcagac
     8701 aaacagaaga acggtatcaa agctaacttc aaaattcgcc ataacgttga agacggtagc
     8761 gtacagctgg cggaccacta ccagcagaac actccgatcg gtgatggtcc ggttctgctg
     8821 ccggataacc actacctgtc cacccagtct aaactgtcca aagacccgaa cgaaaagcgc
     8881 gaccacatgg tgctgctgga gttcgttact gccgcaggta tcacgcacgg catggatgaa
     8941 ctctacaaat aaggtctcag ttttagagct agaaatagca agttaaaata aggctagtcc
     9001 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgagatctg tccataccca
     9061 tggtctagat tgacaattaa tcatccggct cgtataatgt gtggaattgt gagcggataa
     9121 caatttcaca ccctaggccg cggccgcgcg aattcaagcc gaagagaagt aagccatgga
     9181 ccactacctc gacattcgct tgcgaccgga cccggaattt cccccggcgc aactcatgag
     9241 cgtgctcttc ggcaagctcc accaggccct ggtggcacag ggcggggaca ggatcggcgt
     9301 gagcttcccc gacctcgacg aaagccgctc ccggctgggc gagcgcctgc gcattcatgc
     9361 ctcggcggac gaccttcgtg ccctgctcgc ccggccctgg ctggaagggt tgcgggacca
     9421 tctgcaattc ggagaaccgg cagtcgtgcc tcaccccaca ccgtaccgtc aggtcagtcg
     9481 ggttcaggcg aaaagcaatc cggaacgcct gcggcggcgg ctcatgcgcc ggcacgatct
     9541 gagtgaggag gaggctcgga aacgcattcc cgatacggtc gcgagagcct tggacctgcc
     9601 cttcgtcacg ctacgcagcc agagcaccgg acagcacttc cgtctcttca tccgccacgg
     9661 gccgttgcag gtgacggcag aggaaggagg attcacctgt tacgggttga gcaaaggagg
     9721 tttcgttccc tggttctgac tcgaggcatc aacgacaagc cccggctatc ccgatagccg
     9781 gggctttgtc ttgacggtgt acaagaacct ctttactatt ttgtaccccg aaaattggca
     9841 gggcgcacgc tctgtcattt cgtttgtgtg tcgtaagaca atcagtggag ctatagatga
     9901 gtagcgaaac cggtccggtt gcagtggatc cgaccctgcg ccgtcgcatt gaaccgcacg
     9961 agtttgaagt gttctttgat ccgcgcgagc tgcgcaaaga aacttgcctg ctgtacgaga
    10021 ttaactgggg tggccgccat agcatctggc gccataccag ccagaacacc aataagcacg
    10081 tggaagtgaa ttttattgaa aaatttacca ccgagcgcta cttctgccct aatacccgct
    10141 gcagcatcac ctggtttctg agctggagcc cgtgcggcga atgtagtcgc gccattaccg
    10201 agttcctgag ccgctatccg catgtgaccc tgttcatcta catcgcccgt ctgtaccatc
    10261 acgccgatcc gcgcaatcgc caaggtctgc gtgatctgat tagcagcggt gtgaccatcc
    10321 agatcatgac cgaacaagag agcggctatt gctggcgcaa cttcgtgaac tattctccga
    10381 gcaacgaagc ccactggccg cgttatccgc atctgtgggt gcgcctgtat gtgctggagc
    10441 tgtactgcat catcctgggc ctgccgcctt gcctgaatat tctgcgccgt aaacagccgc
    10501 aactgacatt cttcaccatc gcactgcaga gctgccatta tcagcgcctg ccgccgcaca
    10561 ttttatgggc caccggtctg aaaagcggta gtgaaactcc gggcacaagc gaaagcgcaa
    10621 ccccggaaag tgataagaaa tactcaatag gcttagctat cggcacaaat agcgtcggat
    10681 gggcggtgat cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata
    10741 cagaccgcca cagtatcaaa aaaaatctta taggggctct tttatttgac agtggagaga
    10801 cagcggaagc gactcgtctc aaacggacag ctcgtagaag gtatacacgt cggaagaatc
    10861 gtatttgtta tctacaggag attttttcaa atgagatggc gaaagtagat gatagtttct
    10921 ttcatcgact tgaagagtct tttttggtgg aagaagacaa gaagcatgaa cgtcatccta
    10981 tttttggaaa tatagtagat gaagttgctt atcatgagaa atatccaact atctatcatc
    11041 tgcgaaaaaa attggtagat tctactgata aagcggattt gcgcttaatc tatttggcct
    11101 tagcgcatat gattaagttt cgtggtcatt ttttgattga gggagattta aatcctgata
    11161 atagtgatgt ggacaaacta tttatccagt tggtacaaac ctacaatcaa ttatttgaag
    11221 aaaaccctat taacgcaagt ggagtagatg ctaaagcgat tctttctgca cgattgagta
    11281 aatcaagacg attagaaaat ctcattgctc agctccccgg tgagaagaaa aatggcttat
    11341 ttgggaatct cattgctttg tcattgggtt tgacccctaa ttttaaatca aattttgatt
    11401 tggcagaaga tgctaaatta cagctttcaa aagatactta cgatgatgat ttagataatt
    11461 tattggcgca aattggagat caatatgctg atttgttttt ggcagctaag aatttatcag
    11521 atgctatttt actttcagat atcctaagag taaatactga aataactaag gctcccctat
    11581 cagcttcaat gattaaacgc tacgatgaac atcatcaaga cttgactctt ttaaaagctt
    11641 tagttcgaca acaacttcca gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg
    11701 gatatgcagg ttatattgat gggggagcta gccaagaaga attttataaa tttatcaaac
    11761 caattttaga aaaaatggat ggtactgagg aattattggt gaaactaaat cgtgaagatt
    11821 tgctgcgcaa gcaacggacc tttgacaacg gctctattcc ccatcaaatt cacttgggtg
    11881 agctgcatgc tattttgaga agacaagaag acttttatcc atttttaaaa gacaatcgtg
    11941 agaagattga aaaaatcttg acttttcgaa ttccttatta tgttggtcca ttggcgcgtg
    12001 gcaatagtcg ttttgcatgg atgactcgga agtctgaaga aacaattacc ccatggaatt
    12061 ttgaagaagt tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc atgacaaact
    12121 ttgataaaaa tcttccaaat gaaaaagtac taccaaaaca tagtttgctt tatgagtatt
    12181 ttacggttta taacgaattg acaaaggtca aatatgttac tgaaggaatg cgaaa
//
