LOCUS       sequence-341069-       12252 bp    DNA     circular SYN 14-APR-2025
DEFINITION  a modularized single-plasmid cytidine base editing (C-to-T) tool for
            Sinorhizobium meliloti.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12252)
  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 12252)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-HCBE
FEATURES             Location/Qualifiers
     source          1..12252
                     /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"
     promoter        complement(1587..2032)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     terminator      2033..2104
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_RNA        complement(2162..2237)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(2279..2584)
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLGHRENDIKNAINLMFWGI"
     primer_bind     complement(2381..2400)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2926..3585)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3500..3519
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(3639..3645,3646..3663,3664..3669))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(3762..4013)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             complement(4035..4658)
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             complement(join(4872..8945,8946..8948,8949..8975))
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     terminator      9425..9519
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(9618..9637)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             9629..10444
                     /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            10595..10703
                     /note="incP origin of transfer"
     CDS             complement(10714..11376)
                     /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      11377..12148
                     /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 ttttcagtgt gtaatcatcg tttagctcaa tcataccgag agcgccgttt
      661 gctaactcag ccgtgcgttt tttatcgctt tgcagaagtt tttgactttc ttgacggaag
      721 aatgatgtgc ttttgccata gtatgctttg ttaaataaag attcttcgcc ttggtagcca
      781 tcttcagttc cagtgtttgc ttcaaatact aagtatttgt ggcctttatc ttctacgtag
      841 tgaggatctc tcagcgtatg gttgtcgcct gagctgtagt tgccttcatc gatgaactgc
      901 tgtacatttt gatacgtttt tccgtcaccg tcaaagattg atttataatc ctctacaccg
      961 ttgatgttca aagagctgtc tgatgctgat acgttaactt gtgcagttgt cagtgtttgt
     1021 ttgccgtaat gtttaccgga gaaatcagtg tagaataaac ggatttttcc gtcagatgta
     1081 aatgtggctg aacctgacca ttcttgtgtt tggtctttta ggatagaatc atttgcatcg
     1141 aatttgtcgc tgtctttaaa gacgcggcca gcgtttttcc agctgtcaat agaagtttcg
     1201 ccgacttttt gatagaacat gtaaatcgat gtgtcatccg catttttagg atctccggct
     1261 aatgcaaaga cgatgtggta gccgtgatag tttgcgacag tgccgtcagc gttttgtaat
     1321 ggccagctgt cccaaacgtc caggcctttt gcagaagaga tatttttaat tgtggacgaa
     1381 tcaaattcag aaacttgata tttttcattt ttttgctgtt cagggatttg cagcatatca
     1441 tggcgtgtaa tatgggaaat gccgtatgtt tccttatatg gcttttggtt cgtttctttc
     1501 gcaaacgctt gagttgcgcc tcctgccagc agtgcggtag taaaggttaa tactgttgct
     1561 tgttttgcaa actttttgat gttcatcgtt catgtctcct tttttatgta ctgtgttagc
     1621 ggtctgcttc ttccagccct cctgtttgaa gatggcaagt tagttacgca caataaaaaa
     1681 agacctaaaa tatgtaaggg gtgacgccaa agtatacact ttgcccttta cacattttag
     1741 gtcttgcctg ctttatcagt aacaaacccg cgcgatttac ttttcgacct cattctatta
     1801 gactctcgtt tggattgcaa ctggtctatt ttcctctttt gtttgataga aaatcataaa
     1861 aggatttgca gactacgggc ctaaagaact aaaaaatcta tctgtttctt ttcattctct
     1921 gtatttttta tagtttctgt tgcatgggca taaagttgcc tttttaatca caattcagaa
     1981 aatatcataa tatctcattt cactaaataa tagtgaacgg caggtatatg tgcaaataaa
     2041 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2101 tctctctaga tactagagtc acactggccc gggccatggg tatggacaga tctcaaaaaa
     2161 agcaccgact cggtgccact ttttcaagtt gataacggac tagccttatt ttaacttgct
     2221 atttctagct ctaaaactga gaccctgcag actggctgtg tataagggag cctgacattt
     2281 atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggccgagat
     2341 cagccacttc ttccccgata acggacaccg gcacactggc catatcggtg gtcatcatgc
     2401 gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca
     2461 gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca ataatatcac
     2521 tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact
     2581 gcatttcacc agcccctgtt ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg
     2641 acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag acgacgggct
     2701 tcattctgca tggttgtgct taccagaccg gagatattga catcatatat gccttgagca
     2761 actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca tacctctttt
     2821 tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat cagcgcgcaa
     2881 atacgcatac tgttatctgg cttttagtaa gccggatcca gatctttacg ccccgccctg
     2941 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     3001 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     3061 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     3121 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     3181 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     3241 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     3301 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     3361 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     3421 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     3481 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     3541 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     3601 tcctgaaaat ctcgacggat cctaactcaa aatccacaca ttatacgagc cggaagcata
     3661 aagtgtaaag cctggggtgc ctaatgcggc cgcggtctcc aggcaggcac ataccgtgtc
     3721 gacccaatcg tgtcaaccgc gtgccgggga tttgtttaaa ctcacagcat cttgatcttg
     3781 ttctcgccgt tgctgtcctg gatgacgagc gcccacggct tatactccgg ggcgtcgctc
     3841 gtgagcagca tgacgttctc gtccgtcgac tcgtcgtacg cggtatggac caggatgtcc
     3901 gactccggct tgttgccgat gacttcctcg acttcctccg ggagcatcag gatgctctcc
     3961 tggatgacca gctgcttgcc cgtctccttc tcgatgatgt ccgagaggtt ggtgctgccg
     4021 ccgctgccgc gcgagacggc cgggctcttc gtggtgtgga ggatcttgac ctggatcatg
     4081 atgctcagct cgctgcggcg cttctccgcg cgcttcaggg tcttctccag ccagcggttc
     4141 tcgttgagct ggttatggct cgactggatg aagatcttgc ggcagcactg gtagtgctcg
     4201 ctgaccatga cgttcaggcc gacgccgttg tcgcggaggt tccacaggcc gatctggttg
     4261 cgcgcgttct tctcgtaata gagcttgcag gcccagatct tcagggtatg gccgttgccg
     4321 cggagctcct ggttatacca ctccaggatc ttctcggcgc agtccgcgca cggcgaccac
     4381 gacgaatacc agttgatcgt gaactggccc gggttgtcgc ggagatactc ctcgaccttg
     4441 cggatgctga agatctcggc gtggatgccg cgctcggtgc cgctctgcgg cttgttgacc
     4501 gcgtagcccc agaagcaggc gcgacgctcg ccgcggcgct tgagctcgaa gaggacatag
     4561 cagcgatggc tgaccgactt cttgttgttg aagaactgct tcttgaacgt gtagatgtcc
     4621 agcttctcgt ggatgcggac atactccgcg tcggtcatgc ggctgcccga gtacttctcg
     4681 acatacggga ccgggatcat gccgcgcttg ccctcgctgt cctccgcgtt ccaccactgc
     4741 tcctccggct tgtcgcggat gcggaggatg tcgcccttct tgaacggcag atcttcttcg
     4801 tcgttgccgt tgaagtcgaa cagcgcgcgg acatactcgg ccgagccgcc gccgcccgag
     4861 ccgccgccac cgtcgccgcc cagctggctc aggtcgatgc gcgtctcgta gaggccggtg
     4921 atcgactgat ggatgagcgt cgcgtccagg acttccttcg tcgaggtgta gcgcttgcgg
     4981 tcgatcgtgg tgtcgaaata cttgaacgcg gccggggcgc ccaggttggt cagcgtgaag
     5041 agatggatga tgttctccgc ctgctcgcgg atcggcttgt cgcgatgctt gttgtaggcg
     5101 ctgaggacct tgtccaggtt cgcgtcggcc aggatgacgc gcttgctgaa ctccgagatc
     5161 tgctcgatga tctcgtcgag ataatgcttg tgctgctcga cgaacagctg cttctgctcg
     5221 ttgtcctccg ggctgccctt gagcttctcg tagtgcgacg ccagatagag gaagttgacg
     5281 tacttcgacg gcagggcgag ctcgttgccc ttctggagct cgccggcgct agcgagcatg
     5341 cgcttgcggc cgttctccag ctcgaagagc gagtacttcg ggagcttgat gatcaggtcc
     5401 ttcttgactt ccttatagcc cttcgcctcc aggaagtcga tcgggttctt ctcgaacgac
     5461 gagcgctcca tgatcgtgat gccgagcagc tccttgacgc tcttgagctt cttcgacttg
     5521 cccttctcga ccttcgcgac gaccaggacc gagtaggcga ccgtcgggct gtcgaagcca
     5581 ccatacttct tcgggtccca gtccttcttg cgggcgatga gcttgtccga gttgcgcttc
     5641 ggcaggatgc tctccttcga gaagccgccc gtctggacct cggtcttctt gacgatgttg
     5701 acctgcggca tgctgaggac cttgcggacc gtggcgaagt cgcggccctt gtcccagacg
     5761 atctcgcccg tctcgccgtt cgtctcgatg agcgggcgct tgcggatctc gccgttggcg
     5821 agggtgatct cggtcttgaa gaagttcatg atgttcgaat agaagaagta cttcgccgtg
     5881 gccttgccga tctcctgctc gctcttggcg atcatcttgc ggacgtcata gaccttgtag
     5941 tcgccataga cgaactccga ctccagcttc gggtacttct tgatcagcgc ggtgccgacg
     6001 accgcgttga ggtaggcgtc atgggcatgg tgatagttgt tgatctcgcg gaccttatag
     6061 aactggaagt ccttgcggaa gtcgctgacg agcttcgact tcagcgtgat gaccttgacc
     6121 tcgcggatca gcttgtcgtt ctcgtcatac ttggtgttca tgcgcgagtc gaggatctgc
     6181 gcgacgtgct tcgtgatctg gcgcgtctcg accagctggc gcttgatgaa gcccgccttg
     6241 tccagctccg agaggccgcc gcgctccgcc ttggtgaggt tgtcgaactt gcgctgggtg
     6301 atcagcttgg cgttgagcag ctggcgccag tagttcttca tcttcttgac gacttcctcg
     6361 ctcgggacgt tgtccgactt gccgcggttc ttgtccgagc gcgtgaggac cttgttgtcg
     6421 atgctgtcgt ccttcaggaa cgactgcggg acgatatggt cgacgtcata gtcgctgagg
     6481 cggttgatgt ccagctcctg gtcgacgtac atgtcgcggc cgttctggag gtaatacaga
     6541 tagagcttct cgttctgcag ctgggtgttc tcgaccgggt gctccttgag gatctgcgag
     6601 cccagctcct tgatgccttc ctcgatgcgc ttcatgcgct cgcggctgtt cttctggccc
     6661 ttctgcgtgg tctggttctc gcgggccatc tcgatgacga tgttctccgg cttatggcgg
     6721 cccatgacct tgacgagctc gtcgacgacc ttgacggtct gcaggatgcc cttcttgatg
     6781 gccggcgagc cggccaggtt ggcgatgtgt tcgtgcaggc tgtcgccctg gccgctgacc
     6841 tgggccttct ggatgtcttc cttgaacgtg aggctgtcgt cgtggatcag ctgcatgaag
     6901 ttgcggttgg cgaagccgtc gctcttcagg aagtcgagga tggtcttgcc cgactgcttg
     6961 tcgcggatgc cgttgatgag cttgcgcgac aggcggcccc agccggtgta gcggcgacgc
     7021 ttcagctgct tcatgacctt gtcgtcgaag aggtgggcat aggtcttcag gcgctcctcg
     7081 atcatctcgc ggtcctcgaa cagcgtgagg gtcaggacga tgtcctccag gatatcctcg
     7141 ttctcctcgt tgtcgaggaa gtccttgtcc ttgatgatct tgagcaggtc atgataggtg
     7201 cccaggctgg cgttgaagcg gtcctcgacg ccgctgatct cgaccgagtc gaagcactcg
     7261 atcttcttga agtagtcttc cttcagctgc ttgaccgtga ccttgcggtt ggtcttgaag
     7321 agcaggtcga cgatcgcctt cttctgctcg ccgctgagga aggccggctt gcgcatgcct
     7381 tcggtgacat acttgacctt cgtcagctcg ttgtagacgg tgaagtactc atagagcagg
     7441 ctgtgcttcg ggaggacctt ctcgttcggc aggttcttgt cgaagttcgt catgcgctcg
     7501 atgaagctct gggcgctggc gcccttgtcg acgacttcct cgaagttcca cggcgtgatc
     7561 gtctcctcgc tcttgcgggt catccacgcg aagcgcgagt tgccgcgtgc cagcgggccg
     7621 acataatacg ggatgcggaa cgtcaggatc ttctcgatct tctcgcggtt gtccttgagg
     7681 aacggataga agtcttcctg gcgacgcagg atcgcatgca gctcgccgag atggatctgg
     7741 tgcgggatcg agccgttgtc gaaggtgcgc tgcttgcgga gcaggtcctc gcggttgagc
     7801 ttgacgagca gctcctccgt gccgtccatc ttctccagga tcggcttgat gaacttgtag
     7861 aactcttcct ggcttgcgcc gccgtcgatg tagcccgcat agccgttctt gctctggtcg
     7921 aagaagatct ccttatactt ctccgggagc tgctggcgga ccagggcctt caggagcgtg
     7981 aggtcctgat ggtgctcgtc gtagcgcttg atcatgctcg ccgacagcgg ggccttcgtg
     8041 atctcggtgt tgacgcggag gatgtcgctg agcaggatcg cgtccgacag gttcttcgcg
     8101 gccaggaaga ggtccgcata ctggtcgccg atctgggcga gcaggttgtc gaggtcgtcg
     8161 tcgtaggtgt ccttcgacag ctggagcttc gcgtcctcgg ccaggtcgaa gttgctcttg
     8221 aagttcgggg tcaggcccag gctcagggcg atgaggttgc cgaagaggcc gttcttcttc
     8281 tcgcccggga gctgggcgat caggttctcc aggcggcgcg acttgctcag gcgggccgag
     8341 aggatggcct tggcgtcgac gccgctggcg ttgatcgggt tctcctcgaa cagctggtta
     8401 tacgtctgga ccagctggat gaagagcttg tcgacgtccg agttgtccgg gttgaggtcg
     8461 ccctcgatca ggaaatggcc gcggaacttg atcatgtgcg cgagggccag gtagatgagg
     8521 cgcaggtcgg ccttgtcggt gctgtcgacc agcttcttgc ggagatgata gatcgtcggg
     8581 tacttctcgt gatacgcgac ctcgtcgacg atgttgccga agatcggatg gcgctcgtgc
     8641 ttcttatctt cttcgacgag gaagctctcc tccaggcgat ggaagaacga gtcgtcgacc
     8701 ttggccatct cgttcgagaa gatctcctgg agatagcaga tgcggttctt gcgacgcgtg
     8761 tagcggcgac gggcggtgcg cttcaggcgg gtggcctccg ccgtctcgcc gctgtcgaac
     8821 agcagggcgc cgatgaggtt cttcttgatg ctgtggcggt cggtgttgcc caggaccttg
     8881 aacttcttcg acgggacctt atactcgtcg gtgatgaccg cccagccgac gctgttcgtg
     8941 ccgatggcca ggccgatgct gtacttcttg tccatctcga gctccagtgc cctgtcgcct
     9001 ttcttccttc aaagcgatca gtggtcaacc ccggacaggc tccgacgcgc gggctgcggc
     9061 aaaaggcctc tcggccccag acctcggtca ttctgcgcgg cgcgtgtcgt gcctggcaag
     9121 caggaaaggc gatggaaaat gacgacacat tttcaaacct tccgccgtaa tcaggcgcgg
     9181 catcacaccg agtccgagtg cccagttttg aaccattcca aaatggctgg caaaacttca
     9241 ttctcccgaa catcgatcga agtcaagcaa ttgcggaaaa aggacggccg ctgccgccct
     9301 ttttcgatac caaagttggc ctgtcaggcg gccgccacgg cgcgtttcgt catcaccttg
     9361 acgagattgg cgcgataggc ggcgcttgca tgaatatcgg aaagcaggtc ggtacccgct
     9421 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     9481 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggcaattg tgtctcaaaa
     9541 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     9601 ttacataaac agtaatacaa ggggtgttat gagccatatt cagcgtgaaa cgagctgtag
     9661 ccgtccgcgt ctgaacagca acatggatgc ggatctgtat ggctataaat gggcgcgtga
     9721 taacgtgggt cagagcggcg cgaccattta tcgtctgtat ggcaaaccgg atgcgccgga
     9781 actgtttctg aaacatggca aaggcagcgt ggcgaacgat gtgaccgatg aaatggtgcg
     9841 tctgaactgg ctgaccgaat ttatgccgct gccgaccatt aaacatttta ttcgcacccc
     9901 ggatgatgcg tggctgctga ccaccgcgat tccgggcaaa accgcgtttc aggtgctgga
     9961 agaatatccg gatagcggcg aaaacattgt ggatgcgctg gccgtgtttc tgcgtcgtct
    10021 gcatagcatt ccggtgtgca actgcccgtt taacagcgat cgtgtgtttc gtctggccca
    10081 ggcgcagagc cgtatgaaca acggcctggt ggatgcgagc gattttgatg atgaacgtaa
    10141 cggctggccg gtggaacagg tgtggaaaga aatgcataaa ctgctgccgt ttagcccgga
    10201 tagcgtggtg acccacggcg attttagcct ggataacctg attttcgatg aaggcaaact
    10261 gattggctgc attgatgtgg gccgtgtggg cattgcggat cgttatcagg atctggccat
    10321 tctgtggaac tgcctgggcg aatttagccc gagcctgcaa aaacgtctgt ttcagaaata
    10381 tggcattgat aatccggata tgaacaaact gcaatttcat ctgatgctgg atgaattttt
    10441 ctaataatta attgaactac ttttccgctg cataaccctg cttcggggtc attatagcga
    10501 ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg
    10561 tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac
    10621 ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga
    10681 atcctgctct gcgaggctgg ccgtaggccg gccctaccgg cgcggcagcg ttacccgtgt
    10741 cggcggctcc aacggctcgc catcgtccag aaaacacggc tcatcgggca tcggcaggcg
    10801 ctgctgcccg cgccgttccc attcctccgt ttcggtcaag gctggcaggt ctggttccat
    10861 gcccggaatg ccgggctggc tgggcggctc ctcgccgggg ccggtcggta gttgctgctc
    10921 gcccggatac agggtcggga tgcggcgcag gtcgccatgc cccaacagcg attcgtcctg
    10981 gtcgtcgtga tcaaccacca cggcggcact gaacaccgac aggcgcaact ggtcgcgggg
    11041 ctggccccac gccacgcggt cattgaccac gtaggccgac acggtgccgg ggccgttgag
    11101 cttcacgacg gagatccagc gctcggccac caagtccttg actgcgtatt ggaccgtccg
    11161 caaagaacgt ccgatgagct tggaaagagt cttttggctg accaccacgg cgttctggtg
    11221 gcccatctgc gcgacgaggt gatgcagcag cattgccgcc gtgggtttcc tcgcaataag
    11281 cccggcccac gcctcatgcg ctttgcgttc cgtttggacc cagtgaccgg gcttgttctt
    11341 ggcttgaatg ccgatttctc tggactgcgt ggccatgctt atctccatgc ggtaggggtg
    11401 ccgcacggtt gcggcaccat gcgcaatcag ctgcaacttt tcggcagcgc gacaacaatt
    11461 atgcgttgcg taaaagtggc agtcaattac agattttctt taacctacgc aatgagctat
    11521 tgcggggggt gccgcaatga gctgttgcgt accccccttt tttaagttgt tgatttttaa
    11581 gtctttcgca tttcgcccta tatctagttc tttggtgccc aaagaagggc acccctgcgg
    11641 ggttccccca cgccttcggc gcggctcccc ctccggcaaa aagtggcccc tccggggctt
    11701 gttgatcgac tgcgcggcct tcggccttgc ccaaggtggc gctgccccct tggaaccccc
    11761 gcactcgccg ccgtgaggct cggggggcag gcgggcgggc ttcgcccttc gactgccccc
    11821 actcgcatag gcttgggtcg ttccaggcgc gtcaaggcca agccgctgcg cggtcgctgc
    11881 gcgagccttg acccgccttc ctcttggtgt ccaaccggca agcgaagcgc gcaggccgca
    11941 ggccggaggc ttttccccag agaaaattaa aaaaattgat ggggcaaggc cgcaggccgc
    12001 gcagttggag ccggtgggta tgtggtcgaa ggctgggtag ccggtgggca atccctgtgg
    12061 tcaagctcgt gggcaggcgc agcctgtcca tcagcttgtc cagcagggtt gtccacgggc
    12121 cgagcgaagc gagccagccg gtggccgctc gcggccatcg tccacatatc cacgggctgg
    12181 caagggagcg cagcgaccgc gcagggcgaa gcccggagag caagcccgta gggggttact
    12241 ctagggcggc gg
//
