LOCUS       sequence-341070-       12338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a modularized single-plasmid adenosine base editing (A-to-G) tool 
            for Sinorhizobium meliloti.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12338)
  AUTHORS   Wang L, Xiao Y, Wei X, Pan J, Duanmu D
  TITLE     Highly Efficient CRISPR-Mediated Base Editing in Sinorhizobium 
            meliloti.
  JOURNAL   Front Microbiol. 2021 Jun 18;12:686008. doi: 
            10.3389/fmicb.2021.686008. eCollection 2021.
  PUBMED    34220774
REFERENCE   2  (bases 1 to 12338)
  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: "Front 
            Microbiol."; date: "2021-06-18"; volume: "12:686008. doi"; pages: " 
            10.3389/fmicb.2021.686008. eCollection 2021"
COMMENT     Sequence Label: pKm-HABE
FEATURES             Location/Qualifiers
     source          1..12338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..87)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(164..1498,1499..1585))
                     /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
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        complement(1586..2031)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     terminator      2032..2103
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_RNA        complement(2161..2236)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(2278..2583)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(2380..2399)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2925..3584)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3499..3518
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(3638..3644,3645..3662,3663..3668))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(join(3761..7837,7838..7840,7841..7864))
                     /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"
     terminator      9511..9605
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(9704..9723)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             9715..10530
                     /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"
     oriT            10681..10789
                     /note="incP origin of transfer"
     CDS             complement(10800..11462)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      11463..12234
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
       61 tttcgactga gcctttcgtt ttatttgatg cctgccgaat tcggatccgg agtaacagga
      121 gtccaagagc gggtacgtac ccatcggcat ttcttttgcg tttttattta ttaactgtta
      181 attgtccttg ttcaaggatg ctgtctttga caacagatgt tttcttgcct ttgatgttca
      241 gcaggaagct cggcgcaaac gttgattgtt tgtctgcgta gaatcctctg tttgtcatat
      301 agcttgtaat cacgacattg tttcctttcg cttgaggtac agcgaagtgt gagtaagtaa
      361 aggttacatc gttaggatca agatccattt ttaacacaag gccagttttg ttcagcggct
      421 tgtatgggcc agttaaagaa ttagaaacat aaccaagcat gtaaatatcg ttagacgtaa
      481 tgccgtcaat cgtcattttt gatccgcggg agtcagtgaa cagataccat ttgccgttca
      541 ttttaaagac gttcgcgcgt tcaatttcat ctgttactgt gttagatgca atcagcggtt
      601 tcatcacttt tttcagtgtg taatcatcgt ttagctcaat cataccgaga gcgccgtttg
      661 ctaactcagc cgtgcgtttt ttatcgcttt gcagaagttt ttgactttct tgacggaaga
      721 atgatgtgct tttgccatag tatgctttgt taaataaaga ttcttcgcct tggtagccat
      781 cttcagttcc agtgtttgct tcaaatacta agtatttgtg gcctttatct tctacgtagt
      841 gaggatctct cagcgtatgg ttgtcgcctg agctgtagtt gccttcatcg atgaactgct
      901 gtacattttg atacgttttt ccgtcaccgt caaagattga tttataatcc tctacaccgt
      961 tgatgttcaa agagctgtct gatgctgata cgttaacttg tgcagttgtc agtgtttgtt
     1021 tgccgtaatg tttaccggag aaatcagtgt agaataaacg gatttttccg tcagatgtaa
     1081 atgtggctga acctgaccat tcttgtgttt ggtcttttag gatagaatca tttgcatcga
     1141 atttgtcgct gtctttaaag acgcggccag cgtttttcca gctgtcaata gaagtttcgc
     1201 cgactttttg atagaacatg taaatcgatg tgtcatccgc atttttagga tctccggcta
     1261 atgcaaagac gatgtggtag ccgtgatagt ttgcgacagt gccgtcagcg ttttgtaatg
     1321 gccagctgtc ccaaacgtcc aggccttttg cagaagagat atttttaatt gtggacgaat
     1381 caaattcaga aacttgatat ttttcatttt tttgctgttc agggatttgc agcatatcat
     1441 ggcgtgtaat atgggaaatg ccgtatgttt ccttatatgg cttttggttc gtttctttcg
     1501 caaacgcttg agttgcgcct cctgccagca gtgcggtagt aaaggttaat actgttgctt
     1561 gttttgcaaa ctttttgatg ttcatcgttc atgtctcctt ttttatgtac tgtgttagcg
     1621 gtctgcttct tccagccctc ctgtttgaag atggcaagtt agttacgcac aataaaaaaa
     1681 gacctaaaat atgtaagggg tgacgccaaa gtatacactt tgccctttac acattttagg
     1741 tcttgcctgc tttatcagta acaaacccgc gcgatttact tttcgacctc attctattag
     1801 actctcgttt ggattgcaac tggtctattt tcctcttttg tttgatagaa aatcataaaa
     1861 ggatttgcag actacgggcc taaagaacta aaaaatctat ctgtttcttt tcattctctg
     1921 tattttttat agtttctgtt gcatgggcat aaagttgcct ttttaatcac aattcagaaa
     1981 atatcataat atctcatttc actaaataat agtgaacggc aggtatatgt gcaaataaaa
     2041 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     2101 ctctctagat actagagtca cactggcccg ggccatgggt atggacagat ctcaaaaaaa
     2161 gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta
     2221 tttctagctc taaaactgag accctgcaga ctggctgtgt ataagggagc ctgacattta
     2281 tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc
     2341 agccacttct tccccgataa cggacaccgg cacactggcc atatcggtgg tcatcatgcg
     2401 ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag
     2461 cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact
     2521 ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg
     2581 catttcacca gcccctgttc tcgtcagcaa aagagccgtt catttcaata aaccgggcga
     2641 cctcagccat cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt
     2701 cattctgcat ggttgtgctt accagaccgg agatattgac atcatatatg ccttgagcaa
     2761 ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt
     2821 gacatacttc gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa
     2881 tacgcatact gttatctggc ttttagtaag ccggatccag atctttacgc cccgccctgc
     2941 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     3001 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     3061 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     3121 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     3181 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     3241 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     3301 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     3361 atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     3421 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     3481 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     3541 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     3601 cctgaaaatc tcgacggatc ctaactcaaa atccacacat tatacgagcc ggaagcataa
     3661 agtgtaaagc ctggggtgcc taatgcggcc gcggtctcca ggcaggcaca taccgtgtcg
     3721 acccaatcgt gtcaaccgcg tgccggggat ttgtttaaac tcagtcgccg ccgagctggc
     3781 tcaggtcgat gcgcgtctcg tacaggccgg tgatcgactg atggatgagc gtcgcgtcca
     3841 ggacttcctt cgtcgaggtg tagcgcttgc ggtcgatcgt ggtgtcgaaa tacttgaacg
     3901 cggccggggc gcccaggttg gtcagcgtga agagatggat gatgttctcc gcctgctcgc
     3961 ggatcggctt gtcgcgatgc ttgttgtagg cgctgaggac cttgtccagg ttcgcgtcgg
     4021 ccaggatgac gcgcttgctg aactccgaga tctgctcgat gatctcgtcg agataatgct
     4081 tgtgctgctc gacgaacagc tgcttctgct cgttgtcctc cgggctgccc ttgagcttct
     4141 cgtagtgcga cgccagatag aggaagttga cgtacttcga cggcagggcg agctcgttgc
     4201 ccttctggag ctcgccggcg ctagcgagca tgcgcttgcg gccgttctcc agctcgaaga
     4261 gcgagtactt cgggagcttg atgatcaggt ccttcttgac ttccttatag cccttcgcct
     4321 ccaggaagtc gatcgggttc ttctcgaacg acgagcgctc catgatcgtg atgccgagca
     4381 gctccttgac gctcttgagc ttcttcgact tgcccttctc gaccttcgcg acgaccagga
     4441 ccgagtaggc gaccgtcggg ctgtcgaagc caccatactt cttcgggtcc cagtccttct
     4501 tgcgggcgat gagcttgtcc gagttgcgct tcggcaggat gctctccttc gagaagccgc
     4561 ccgtctggac ctcggtcttc ttgacgatgt tgacctgcgg catgctgagg accttgcgga
     4621 ccgtggcgaa gtcgcggccc ttgtcccaga cgatctcgcc cgtctcgccg ttcgtctcga
     4681 tgagcgggcg cttgcggatc tcgccgttgg cgagggtgat ctcggtcttg aagaagttca
     4741 tgatgttcga atagaagaag tacttcgccg tggccttgcc gatctcctgc tcgctcttgg
     4801 cgatcatctt gcggacgtca tagaccttgt agtcgccata gacgaactcc gactccagct
     4861 tcgggtactt cttgatcagc gcggtgccga cgaccgcgtt gaggtaggcg tcatgggcat
     4921 ggtgatagtt gttgatctcg cggaccttat agaactggaa gtccttgcgg aagtcgctga
     4981 cgagcttcga cttcagcgtg atgaccttga cctcgcggat cagcttgtcg ttctcgtcat
     5041 acttggtgtt catgcgcgag tcgaggatct gcgcgacgtg cttcgtgatc tggcgcgtct
     5101 cgaccagctg gcgcttgatg aagcccgcct tgtccagctc cgagaggccg ccgcgctccg
     5161 ccttggtgag gttgtcgaac ttgcgctggg tgatcagctt ggcgttgagc agctggcgcc
     5221 agtagttctt catcttcttg acgacttcct cgctcgggac gttgtccgac ttgccgcggt
     5281 tcttgtccga gcgcgtgagg accttgttgt cgatgctgtc gtccttcagg aacgactgcg
     5341 ggacgatatg gtcgacgtca tagtcgctga ggcggttgat gtccagctcc tggtcgacgt
     5401 acatgtcgcg gccgttctgg aggtaataca gatagagctt ctcgttctgc agctgggtgt
     5461 tctcgaccgg gtgctccttg aggatctgcg agcccagctc cttgatgcct tcctcgatgc
     5521 gcttcatgcg ctcgcggctg ttcttctggc ccttctgcgt ggtctggttc tcgcgggcca
     5581 tctcgatgac gatgttctcc ggcttatggc ggcccatgac cttgacgagc tcgtcgacga
     5641 ccttgacggt ctgcaggatg cccttcttga tggccggcga gccggccagg ttggcgatgt
     5701 gttcgtgcag gctgtcgccc tggccgctga cctgggcctt ctggatgtct tccttgaacg
     5761 tgaggctgtc gtcgtggatc agctgcatga agttgcggtt ggcgaagccg tcgctcttca
     5821 ggaagtcgag gatggtcttg cccgactgct tgtcgcggat gccgttgatg agcttgcgcg
     5881 acaggcggcc ccagccggtg tagcggcgac gcttcagctg cttcatgacc ttgtcgtcga
     5941 agaggtgggc ataggtcttc aggcgctcct cgatcatctc gcggtcctcg aacagcgtga
     6001 gggtcaggac gatgtcctcc aggatatcct cgttctcctc gttgtcgagg aagtccttgt
     6061 ccttgatgat cttgagcagg tcatgatagg tgcccaggct ggcgttgaag cggtcctcga
     6121 cgccgctgat ctcgaccgag tcgaagcact cgatcttctt gaagtagtct tccttcagct
     6181 gcttgaccgt gaccttgcgg ttggtcttga agagcaggtc gacgatcgcc ttcttctgct
     6241 cgccgctgag gaaggccggc ttgcgcatgc cttcggtgac atacttgacc ttcgtcagct
     6301 cgttgtagac ggtgaagtac tcatagagca ggctgtgctt cgggaggacc ttctcgttcg
     6361 gcaggttctt gtcgaagttc gtcatgcgct cgatgaagct ctgggcgctg gcgcccttgt
     6421 cgacgacttc ctcgaagttc cacggcgtga tcgtctcctc gctcttgcgg gtcatccacg
     6481 cgaagcgcga gttgccgcgt gccagcgggc cgacataata cgggatgcgg aacgtcagga
     6541 tcttctcgat cttctcgcgg ttgtccttga ggaacggata gaagtcttcc tggcgacgca
     6601 ggatcgcatg cagctcgccg agatggatct ggtgcgggat cgagccgttg tcgaaggtgc
     6661 gctgcttgcg gagcaggtcc tcgcggttga gcttgacgag cagctcctcc gtgccgtcca
     6721 tcttctccag gatcggcttg atgaacttgt agaactcttc ctggcttgcg ccgccgtcga
     6781 tgtagcccgc atagccgttc ttgctctggt cgaagaagat ctccttatac ttctccggga
     6841 gctgctggcg gaccagggcc ttcaggagcg tgaggtcctg atggtgctcg tcgtagcgct
     6901 tgatcatgct cgccgacagc ggggccttcg tgatctcggt gttgacgcgg aggatgtcgc
     6961 tgagcaggat cgcgtccgac aggttcttcg cggccaggaa gaggtccgca tactggtcgc
     7021 cgatctgggc gagcaggttg tcgaggtcgt cgtcgtaggt gtccttcgac agctggagct
     7081 tcgcgtcctc ggccaggtcg aagttgctct tgaagttcgg ggtcaggccc aggctcaggg
     7141 cgatgaggtt gccgaagagg ccgttcttct tctcgcccgg gagctgggcg atcaggttct
     7201 ccaggcggcg cgacttgctc aggcgggccg agaggatggc cttggcgtcg acgccgctgg
     7261 cgttgatcgg gttctcctcg aacagctggt tatacgtctg gaccagctgg atgaagagct
     7321 tgtcgacgtc cgagttgtcc gggttgaggt cgccctcgat caggaaatgg ccgcggaact
     7381 tgatcatgtg cgcgagggcc aggtagatga ggcgcaggtc ggccttgtcg gtgctgtcga
     7441 ccagcttctt gcggagatga tagatcgtcg ggtacttctc gtgatacgcg acctcgtcga
     7501 cgatgttgcc gaagatcgga tggcgctcgt gcttcttatc ttcttcgacg aggaagctct
     7561 cctccaggcg atggaagaac gagtcgtcga ccttggccat ctcgttcgag aagatctcct
     7621 ggagatagca gatgcggttc ttgcgacgcg tgtagcggcg acgggcggtg cgcttcaggc
     7681 gggtggcctc cgccgtctcg ccgctgtcga acagcagggc gccgatgagg ttcttcttga
     7741 tgctgtggcg gtcggtgttg cccaggacct tgaacttctt cgacgggacc ttatactcgt
     7801 cggtgatgac cgcccagccg acgctgttcg tgccgatggc caggccgatg ctgtacttct
     7861 tgtctccgat gctgccgccc gacgagccgc ccgaggactc cggcgtggcg ctctccgacg
     7921 tgccaggcgt ctccgacccc gacgagccgc ccgacgagcc gcccgaatcc gtcgagctct
     7981 gcgccttctt ctgcgcgttg aagacctggc gcggcatgcg gaagaagtag cagagcagcg
     8041 cggcgcattc atcggccagg atgccttcgg tgatttccac ccggtgattc atgcccgggt
     8101 agtggagcac atccatcagg ctgccggccg cgcccgtctt cgcattgcgg acgccgaaga
     8161 ccacccggcc gatgcgcgaa tggatcatcg cgcccgcgca catcacgcac ggttcgaagg
     8221 tgacgtagag ggtggcgtcg atcaggcgat agttttgcat gacgaggccg ccctgccgga
     8281 gggccatgat ttcggcgtgc gcggtcggat catgcaggcc gatggcgcgg ttccagcctt
     8341 cgccgatcac ccgattattc aggaccagga ccgcgccgac cgggacctcc cgttcgtcgc
     8401 gcgcgcgctt ggccagcgtc agggcgtgcc gcatccagta ctcgtggctg aactccactt
     8461 ccgacgagcc gcccgacgag ccgcccgagc tctccggcgt cgcgctctcc gacgtgcccg
     8521 gcgtctcgct acccgacgag ccgcccgacg agccgcccga gtcggtgctc gactgcgcct
     8581 tcttctgggc cttgatctcc tggcgacgca tgcggaagaa gtccgagagc agcgcggcgc
     8641 actcgtccgc caggatgccc tcggtgatct cgacgcgatg gttcatgccc ggatggtgga
     8701 ggacgtccat caggctgccc gcggcgcccg tcttggcgtc gcgggcgccg aagacgacgc
     8761 ggccgatgcg cgagtggatc atggcgcccg cgcacatgac gcacggctcc agggtgacgt
     8821 acagggtcgc gtcgatgagg cgatagttct gcatgacgag gccgccctgg cgcagggcca
     8881 tgatctcggc atgggcggtc gggtcatggc ggccgatcgg gcggttccag ccctcgccga
     8941 tgacgcggtt gttgtggacc aggacggcgc cgaccgggac ctcgcgctcg tcccacgcgc
     9001 gcttggcgag ggtcagggca tggcgcatcc aatactcgtg gctgaactcg acctccgaca
     9061 tctcgagctc cagtgccctg tcgcctttct tccttcaaag cgatcagtgg tcaaccccgg
     9121 acaggctccg acgcgcgggc tgcggcaaaa ggcctctcgg ccccagacct cggtcattct
     9181 gcgcggcgcg tgtcgtgcct ggcaagcagg aaaggcgatg gaaaatgacg acacattttc
     9241 aaaccttccg ccgtaatcag gcgcggcatc acaccgagtc cgagtgccca gttttgaacc
     9301 attccaaaat ggctggcaaa acttcattct cccgaacatc gatcgaagtc aagcaattgc
     9361 ggaaaaagga cggccgctgc cgcccttttt cgataccaaa gttggcctgt caggcggccg
     9421 ccacggcgcg tttcgtcatc accttgacga gattggcgcg ataggcggcg cttgcatgaa
     9481 tatcggaaag caggtcggta cccgctcttg gactcctgtt gatagatcca gtaatgacct
     9541 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     9601 agaatccagg caattgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     9661 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     9721 catattcagc gtgaaacgag ctgtagccgt ccgcgtctga acagcaacat ggatgcggat
     9781 ctgtatggct ataaatgggc gcgtgataac gtgggtcaga gcggcgcgac catttatcgt
     9841 ctgtatggca aaccggatgc gccggaactg tttctgaaac atggcaaagg cagcgtggcg
     9901 aacgatgtga ccgatgaaat ggtgcgtctg aactggctga ccgaatttat gccgctgccg
     9961 accattaaac attttattcg caccccggat gatgcgtggc tgctgaccac cgcgattccg
    10021 ggcaaaaccg cgtttcaggt gctggaagaa tatccggata gcggcgaaaa cattgtggat
    10081 gcgctggccg tgtttctgcg tcgtctgcat agcattccgg tgtgcaactg cccgtttaac
    10141 agcgatcgtg tgtttcgtct ggcccaggcg cagagccgta tgaacaacgg cctggtggat
    10201 gcgagcgatt ttgatgatga acgtaacggc tggccggtgg aacaggtgtg gaaagaaatg
    10261 cataaactgc tgccgtttag cccggatagc gtggtgaccc acggcgattt tagcctggat
    10321 aacctgattt tcgatgaagg caaactgatt ggctgcattg atgtgggccg tgtgggcatt
    10381 gcggatcgtt atcaggatct ggccattctg tggaactgcc tgggcgaatt tagcccgagc
    10441 ctgcaaaaac gtctgtttca gaaatatggc attgataatc cggatatgaa caaactgcaa
    10501 tttcatctga tgctggatga atttttctaa taattaattg aactactttt ccgctgcata
    10561 accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata
    10621 tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc
    10681 gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta
    10741 ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgt aggccggccc
    10801 taccggcgcg gcagcgttac ccgtgtcggc ggctccaacg gctcgccatc gtccagaaaa
    10861 cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc gttcccattc ctccgtttcg
    10921 gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg gctggctggg cggctcctcg
    10981 ccggggccgg tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg gcgcaggtcg
    11041 ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc ggcactgaac
    11101 accgacaggc gcaactggtc gcggggctgg ccccacgcca cgcggtcatt gaccacgtag
    11161 gccgacacgg tgccggggcc gttgagcttc acgacggaga tccagcgctc ggccaccaag
    11221 tccttgactg cgtattggac cgtccgcaaa gaacgtccga tgagcttgga aagagtcttt
    11281 tggctgacca ccacggcgtt ctggtggccc atctgcgcga cgaggtgatg cagcagcatt
    11341 gccgccgtgg gtttcctcgc aataagcccg gcccacgcct catgcgcttt gcgttccgtt
    11401 tggacccagt gaccgggctt gttcttggct tgaatgccga tttctctgga ctgcgtggcc
    11461 atgcttatct ccatgcggta ggggtgccgc acggttgcgg caccatgcgc aatcagctgc
    11521 aacttttcgg cagcgcgaca acaattatgc gttgcgtaaa agtggcagtc aattacagat
    11581 tttctttaac ctacgcaatg agctattgcg gggggtgccg caatgagctg ttgcgtaccc
    11641 ccctttttta agttgttgat ttttaagtct ttcgcatttc gccctatatc tagttctttg
    11701 gtgcccaaag aagggcaccc ctgcggggtt cccccacgcc ttcggcgcgg ctccccctcc
    11761 ggcaaaaagt ggcccctccg gggcttgttg atcgactgcg cggccttcgg ccttgcccaa
    11821 ggtggcgctg cccccttgga acccccgcac tcgccgccgt gaggctcggg gggcaggcgg
    11881 gcgggcttcg cccttcgact gcccccactc gcataggctt gggtcgttcc aggcgcgtca
    11941 aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc gccttcctct tggtgtccaa
    12001 ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt ccccagagaa aattaaaaaa
    12061 attgatgggg caaggccgca ggccgcgcag ttggagccgg tgggtatgtg gtcgaaggct
    12121 gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc aggcgcagcc tgtccatcag
    12181 cttgtccagc agggttgtcc acgggccgag cgaagcgagc cagccggtgg ccgctcgcgg
    12241 ccatcgtcca catatccacg ggctggcaag ggagcgcagc gaccgcgcag ggcgaagccc
    12301 ggagagcaag cccgtagggg gttactctag ggcggcgg
//
