LOCUS       sequence-468502-       11668 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses arabinose-inducible dCas9, codon-optimized for Salmonella.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11668)
  AUTHORS   Naseri G, Raasch H, Charpentier E, Erhardt M
  TITLE     A versatile regulatory toolkit of arabinose-inducible artificial 
            transcription factors for Enterobacteriaceae.
  JOURNAL   Commun Biol. 2023 Oct 3;6(1):1005. doi: 10.1038/s42003-023-05363-3.
  PUBMED    37789111
REFERENCE   2  (bases 1 to 11668)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2023-10-3"; volume: "6(1):1005. doi"; pages: " 
            10.1038/s42003-023-05363-3"
COMMENT     Sequence Label: pG0A0-1-1-PBAD-dCas9
FEATURES             Location/Qualifiers
     source          1..11668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     330..349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(554..1357)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1358..1573)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      2174..3051
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     3222..3239
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(3397..4191)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3397..4191)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3508..3527)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4118..4137
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4192..4296)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4451..4473
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4551..4628
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4629..5711
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4648..4667)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5943..5948,5949..5965,5966..5972)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(5976..6854)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        6881..7165
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     7088..7107
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             7195..7206
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(7213..7239,7240..7242,7243..9729,9730..9732,
                     9733..11319)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     terminator      11343..11414
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      11430..11457
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      join(11509..11668,1..429)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
       61 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
      121 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
      181 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
      241 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
      301 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
      361 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
      421 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      481 gctcatgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
      541 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
      601 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
      661 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
      721 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
      781 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
      841 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
      901 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
      961 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
     1021 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
     1081 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
     1141 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
     1201 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     1261 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     1321 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg
     1381 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
     1441 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
     1501 agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa
     1561 atgatgaatt gaaaagctag cttatcgatg ataagctgtc aaacatgaga attaattcca
     1621 cggactatag actataccta gtatactccg tctactgtac gatacacttc cgctcaggtc
     1681 cttgtccttt aacgaggcct taccactctt ttgttactct attgatccag ctcagcaaag
     1741 gcagtgtgat ctaagattct atcttcgcga tgtagtaaaa ctagctagac cgagaaagag
     1801 actagaaatg caaaaggcac ttctacaatg gctgccatca ttattatccg atgtgacgct
     1861 gcagcttctc aatgatattc gaatacgctt tgaggagata cagcctaata tccgacaaac
     1921 tgttttacag atttacgatc gtacttgtta cccatcattg aattttgaac atccgaacct
     1981 gggagttttc cctgaaacag atagtatatt tgaacctgta taataatata tagtctagcg
     2041 ctttacggaa gacaatgtat gtatttcggt tcctggagaa actattgcat ctattgcata
     2101 ggtaatcttg cacgtcgcat ccccggttca ttttctgcgt ttccatcttg cacttcaata
     2161 gcatatcttt gttaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     2221 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     2281 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     2341 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2401 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     2461 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     2521 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     2581 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     2641 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     2701 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     2761 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     2821 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     2881 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     2941 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     3001 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tgggaagctc
     3061 caccccggtt gataatcaga aaagccccaa aaacaggaag attgtataag caaatattta
     3121 aattgtaatt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3181 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3241 aagcggaaga gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3301 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3361 aatctaaagt atatatgagt aaacttggtc tgacagtcag aagaactcgt caagaaggcg
     3421 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     3481 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     3541 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     3601 catgatattc ggcaagcagg catcgccatg agtcacgacg agatcctcgc cgtcgggcat
     3661 gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     3721 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     3781 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     3841 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     3901 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     3961 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     4021 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     4081 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     4141 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tactcttcct
     4201 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4261 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4321 tgacgtcgga cggatcgctt gcctgtaact tacacgcgcc tcgtggggcg cgccccgcca
     4381 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4441 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4501 aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg catttacgtt gacaccatcg
     4561 aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
     4621 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
     4681 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
     4741 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
     4801 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     4861 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     4921 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     4981 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     5041 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     5101 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     5161 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     5221 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     5281 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     5341 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     5401 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     5461 acagctcatg ttatatcccg ccgtcaacca ccatcaaaca ggattttcgc ctgctggggc
     5521 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     5581 tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
     5641 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5701 gcgggcagtg agcgcaacgc aattaatgtg agttagcgcg aattgatctg gtttgacagc
     5761 ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
     5821 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     5881 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
     5941 tgttgacaat taatcatccg gctcgtataa tgtgtttatg acaacttgac ggctacatca
     6001 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
     6061 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
     6121 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
     6181 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
     6241 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
     6301 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
     6361 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
     6421 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
     6481 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
     6541 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
     6601 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
     6661 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
     6721 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
     6781 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
     6841 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     6901 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     6961 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     7021 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     7081 ctttgctatg ccatagcatt tttatccata agattagcgg attctacctg acgcttttta
     7141 tcgcaactct ctactgtttc tccatacccg tttttttggg aattcaaaag atctaaagag
     7201 gagaaaggat ctatggataa gaagtactcc attggtttag caattggcac gaacagtgtc
     7261 ggatgggccg tgattacaga tgaatacaag gtcccttcaa agaaattcaa agtgttaggt
     7321 aatactgacc ggcacagtat taagaaaaac ctgatcgggg cacttctgtt cgattccggg
     7381 gaaactgcag aggcaacacg tttgaagcgt accgcccggc gccgttatac gcgtcgtaaa
     7441 aatcgcatct gttacttaca agaaatcttc tcaaatgaga tggcgaaggt agacgattct
     7501 ttttttcatc ggctggagga atcgtttttg gtagaagagg acaagaagca tgaacgccat
     7561 ccgattttcg ggaatattgt cgatgaagtt gcctatcatg agaaatatcc gacaatttac
     7621 catttacgga aaaaacttgt ggattctacg gacaaagctg acttgcggtt gatctatctt
     7681 gccctggccc acatgattaa gttccgtggg cactttctta tcgaagggga tttgaaccca
     7741 gataactcgg acgttgacaa gttgtttatt caattggtcc aaacctataa ccagcttttc
     7801 gaggaaaatc cgatcaatgc tagcggtgtg gacgcgaaag ctatcctgag tgcccggtta
     7861 tctaaatcac ggcggctgga gaacttgatt gcccagttgc ccggagagaa gaaaaacgga
     7921 ttattcggta atcttatcgc cctttctctg gggttgacgc ccaacttcaa atctaacttt
     7981 gatctggccg aagacgctaa attacaactt agtaaagata cctacgacga tgacctggat
     8041 aatttgttgg ctcaaatcgg ggatcaatat gctgaccttt tccttgcggc gaagaactta
     8101 tcggacgcca tcttgctgtc ggatatcctg cgtgtcaata cggagattac taaggcacca
     8161 ctttccgcat ccatgatcaa gcgttatgac gagcatcacc aagaccttac gttgctgaaa
     8221 gccttagtcc gtcaacagtt acctgaaaaa tataaagaaa ttttttttga tcagtctaag
     8281 aatggatacg caggttatat cgacggagga gcaagccaag aagaatttta caaatttatt
     8341 aaaccgattc ttgaaaagat ggatggtacg gaggaacttc tggttaagtt aaatcgcgaa
     8401 gacttgttgc gcaagcagcg cactttcgac aacgggagta tcccccacca aattcatctt
     8461 ggtgaattac atgcgatcct gcggcggcaa gaagattttt atcctttctt aaaagataat
     8521 cgcgaaaaaa ttgaaaaaat cttaacgttt cgcattcctt attacgtggg ccctctggca
     8581 cgcggtaata gccggtttgc ctggatgacc cgcaaatcag aagagaccat caccccgtgg
     8641 aatttcgagg aggtagtaga caagggtgcc agtgcacaat ccttcatcga acgcatgaca
     8701 aattttgata aaaatttgcc taatgagaag gtcctgccga aacatagtct tttatatgaa
     8761 tactttaccg tatacaacga gttgacaaag gttaagtacg tgacggaggg gatgcgtaaa
     8821 cctgcatttt tgtcaggtga gcaaaaaaaa gcgattgtgg atttattatt taaaactaat
     8881 cggaaggtga cagtgaagca acttaaggaa gactatttca agaagatcga atgcttcgat
     8941 tcggtagaaa tcagtggtgt tgaagaccgc ttcaacgcaa gcttagggac ttatcacgat
     9001 cttcttaaga tcattaagga caaagacttc ctggacaacg aggaaaacga ggacattttg
     9061 gaagatattg tacttacttt gactttgttt gaagaccggg agatgatcga agagcgtctt
     9121 aagacgtacg cgcacctttt cgatgacaaa gtaatgaagc agctgaagcg gcggcggtac
     9181 accggatggg gtcgtttatc tcggaaattg atcaatggca ttcgtgacaa gcaaagcggc
     9241 aagacgatct tagatttcct taagtccgat ggatttgcta atcggaactt tatgcaattg
     9301 attcatgacg atagtcttac ctttaaagag gacatccaaa aggcacaagt gtcaggccaa
     9361 ggtgacagtc ttcacgaaca tattgctaac ttagccggta gtccagctat caagaagggt
     9421 atcttacaga ctgttaaagt agtagatgaa ctggtgaaag taatgggtcg tcacaagcct
     9481 gagaatatcg taattgaaat ggcgcgggaa aatcagacaa ctcaaaaagg tcaaaaaaac
     9541 agtcgtgaac gtatgaaacg cattgaagag ggcatcaagg agttgggcag tcagatttta
     9601 aaggagcatc cagtggagaa cactcagctg caaaatgaga aactttatct ttattacctt
     9661 cagaatggac gcgatatgta cgtcgatcaa gaactggata ttaatcggtt atccgattac
     9721 gacgttgatg ccatcgtccc tcaatcattt ctgaaagacg atagcatcga caataaggtg
     9781 ctgacacgta gtgacaagaa ccgtgggaag agcgacaacg taccgtcgga ggaggtcgtt
     9841 aagaagatga agaactattg gcggcaattg ctgaacgcta aattgattac gcagcggaaa
     9901 tttgacaacc tgaccaaggc agagcgtggg ggcctgagtg aactggataa agctggcttt
     9961 atcaaacgtc aattggtaga gacacgccaa attacgaagc atgtcgcgca gattttggac
    10021 tcgcgcatga acactaagta cgatgaaaac gacaagttaa ttcgtgaggt gaaagtcatt
    10081 actcttaaga gtaaacttgt ctcggacttc cggaaggact ttcaatttta caaagtccgt
    10141 gaaattaata actatcatca cgcccacgac gcatatctta acgcggtcgt tggaacagct
    10201 ttgattaaga aatatcccaa gcttgagagc gagttcgtgt acggtgatta taaagtgtat
    10261 gacgtacgta agatgatcgc gaagtccgaa caagagattg ggaaggcaac tgctaagtac
    10321 tttttttact ctaatattat gaactttttc aaaaccgaga tcacattggc taatggcgaa
    10381 atccgtaagc gccctcttat tgagacaaac ggggaaactg gggagatcgt gtgggataag
    10441 ggccgcgatt ttgcaactgt ccggaaggtc ctgtctatgc cccaagtgaa catcgttaag
    10501 aagacggaag ttcagaccgg gggcttttca aaggagagta ttttacccaa gcgcaacagt
    10561 gacaagttga tcgcgcggaa gaaggactgg gatcccaaaa agtatggggg attcgactca
    10621 ccgaccgttg catactccgt gctggtcgtc gcgaaagtgg agaagggcaa gagtaaaaaa
    10681 ctgaagtcgg ttaaggagtt gcttggcatc actattatgg aacgtagtag cttcgagaag
    10741 aatccaatcg attttttaga agccaagggc tacaaagaag ttaagaaaga ccttattatc
    10801 aagctgccaa agtactcact tttcgaatta gaaaacggac gtaagcgcat gttagcgtcc
    10861 gctggtgaac tgcaaaaagg aaatgagctg gcactgcctt ctaaatacgt gaacttttta
    10921 tatttggcgt cccattacga aaagcttaag gggtcgcccg aagataacga gcaaaagcag
    10981 ctgtttgtgg aacaacacaa gcattatctg gatgagatca tcgagcagat tagcgaattc
    11041 tcaaagcggg tgatcttagc cgatgctaac ttggataaag tgctgtccgc ttacaacaaa
    11101 catcgcgata agcccatccg tgagcaggcg gagaatatca ttcatctgtt tacattgacg
    11161 aatttggggg cccccgctgc attcaagtac ttcgatacta caatcgaccg taaacgttac
    11221 actagtacaa aagaagtttt ggacgctacc ctgatccatc agagtatcac tggcctgtac
    11281 gagacacgca tcgacttgtc ccaattgggg ggggattaac tcgagtaagg atctccaggc
    11341 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
    11401 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
    11461 ataaaaccct gcaggggccg gccccgtaga aaagatcaaa ggatcttctt gagatccttt
    11521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
    11581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
    11641 gataccaaat actgttcttc tagtgtag
//
