LOCUS       sequence-436148-       14077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coxiella burnetii CRISPRi plasmid without sgRNA construct. Expresses
            3xF-dCas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14077)
  AUTHORS   Steiner S, Roy CR
  TITLE     CRISPR-Cas9-based approaches for genetic analysis and epistatic 
            interaction studies in Coxiella burnetii.
  JOURNAL   mSphere. 2024 Nov 19:e0052324. doi: 10.1128/msphere.00523-24.
  PUBMED    39560384
REFERENCE   2  (bases 1 to 14077)
  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: "mSphere. 
            2024 Nov 19:e0052324. doi: 10.1128/msphere.00523-24."
COMMENT     Sequence Label: pCB-CRISPRi
FEATURES             Location/Qualifiers
     source          1..14077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             211..276
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(277..303,304..306,307..2793,2794..2796,2797..4383)
                     /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"
     CDS             complement(4403..5197)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVSRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDRDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4514..4533)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5124..5143
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5437..5454)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5607..5693
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5785..5812
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5831..5922
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6165..6184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7244..7638
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     CDS             9183..10154
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             10665..11504
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             11491..12342
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(12619..13701)
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     13663..13682
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(13702..13779)
                     /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."
     promoter        join(14009..14014,14015..14030,14031..14037)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    14045..14061
                     /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)."
     primer_bind     14050..14072
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 gaattcatgg cttcgtttcg cagcgaactt ggaaaaaaaa tttttggcga ttcgccgctt
       61 tcgtataacg aagtaaaaac ctatctcagc gaacacccag gcggaaccgc agaacgagcc
      121 tggaatcata tttgtgcgcc ttgaaatatc cctctccata ccccataaat aaataatgac
      181 attaatcctt catgaaggag gctgcagatg gactacaaag accatgacgg tgattataaa
      241 gatcatgata tcgattacaa ggatgacgat gacaagatgg acaagaagta ttctatcgga
      301 ctggccatcg ggactaatag cgtcgggtgg gccgtgatca ctgacgagta caaggtgccc
      361 tctaagaagt tcaaggtgct cgggaacacc gaccggcatt ccatcaagaa aaatctgatc
      421 ggagctctcc tctttgattc aggggagacc gctgaagcaa cccgcctcaa gcggactgct
      481 agacggcggt acaccaggag gaagaaccgg atttgttacc ttcaagagat attctccaac
      541 gaaatggcaa aggtcgacga cagcttcttc cataggctgg aagaatcatt cctcgtggaa
      601 gaggataaga agcatgaacg gcatcccatc ttcggtaata tcgtcgacga ggtggcctat
      661 cacgagaaat acccaaccat ctaccatctt cgcaaaaagc tggtggactc aaccgacaag
      721 gcagacctcc ggcttatcta cctggccctg gcccacatga tcaagttcag aggccacttc
      781 ctgatcgagg gcgacctcaa tcctgacaat agcgatgtgg ataaactgtt catccagctg
      841 gtgcagactt acaaccagct ctttgaagag aaccccatca atgcaagcgg agtcgatgcc
      901 aaggccattc tgtcagcccg gctgtcaaag agccgcagac ttgagaatct tatcgctcag
      961 ctgccgggtg aaaagaaaaa tggactgttc gggaacctga ttgctctttc acttgggctg
     1021 actcccaatt tcaagtctaa tttcgacctg gcagaggatg ccaagctgca actgtccaag
     1081 gacacctatg atgacgatct cgacaacctc ctggcccaga tcggtgacca atacgccgac
     1141 cttttccttg ctgctaagaa tctttctgac gccatcctgc tgtctgacat tctccgcgtg
     1201 aacactgaaa tcaccaaggc ccctctttca gcttcaatga ttaagcggta tgatgagcac
     1261 caccaggacc tgaccctgct taaggcactc gtccggcagc agcttccgga gaagtacaag
     1321 gaaatcttct ttgaccagtc aaagaatgga tacgccggct acatcgacgg aggtgcctcc
     1381 caagaggaat tttataagtt tatcaaacct atccttgaga agatggacgg caccgaagag
     1441 ctcctcgtga aactgaatcg ggaggatctg ctgcggaagc agcgcacttt cgacaatggg
     1501 agcattcccc accagatcca tcttggggag cttcacgcca tccttcggcg ccaagaggac
     1561 ttctacccct ttcttaagga caacagggag aagattgaga aaattctcac tttccgcatc
     1621 ccctactacg tgggacccct cgccagagga aatagccggt ttgcttggat gaccagaaag
     1681 tcagaagaaa ctatcactcc ctggaacttc gaagaggtgg tggacaaggg agccagcgct
     1741 cagtcattca tcgaacggat gactaacttc gataagaacc tccccaatga gaaggtcctg
     1801 ccgaaacatt ccctgctcta cgagtacttt accgtgtaca acgagctgac caaggtgaaa
     1861 tatgtcaccg aagggatgag gaagcccgca ttcctgtcag gcgaacaaaa gaaggcaatt
     1921 gtggaccttc tgttcaagac caatagaaag gtgaccgtga agcagctgaa ggaggactat
     1981 ttcaagaaaa ttgaatgctt cgactctgtg gagattagcg gggtcgaaga tcggttcaac
     2041 gcaagcctgg gtacctacca tgatctgctt aagatcatca aggacaagga ttttctggac
     2101 aatgaggaga acgaggacat ccttgaggac attgtcctga ctctcactct gttcgaggac
     2161 cgggaaatga tcgaggagag gcttaagacc tacgcccatc tgttcgacga taaagtgatg
     2221 aagcaactta aacggagaag atataccgga tggggacgcc ttagccgcaa actcatcaac
     2281 ggaatccggg acaaacagag cggaaagacc attcttgatt tccttaagag cgacggattc
     2341 gctaatcgca acttcatgca acttatccat gatgattccc tgacctttaa ggaggacatc
     2401 cagaaggccc aagtgtctgg acaaggtgac tcactgcacg agcatatcgc aaatctggct
     2461 ggttcacccg ctattaagaa gggtattctc cagaccgtga aagtcgtgga cgagctggtc
     2521 aaggtgatgg gtcgccataa accagagaac attgtcatcg agatggccag ggaaaaccag
     2581 actacccaga agggacagaa gaacagcagg gagcggatga aaagaattga ggaagggatt
     2641 aaggagctcg ggtcacagat ccttaaagag cacccggtgg aaaacaccca gcttcagaat
     2701 gagaagctct atctgtacta ccttcaaaat ggacgcgata tgtatgtgga ccaagagctt
     2761 gatatcaaca ggctctcaga ctacgacgtg gacgccatcg tccctcagag cttcctcaaa
     2821 gacgactcaa ttgacaataa ggtgctgact cgctcagaca agaaccgggg aaagtcagat
     2881 aacgtgccct cagaggaagt cgtgaaaaag atgaagaact attggcgcca gcttctgaac
     2941 gcaaagctga tcactcagcg gaagttcgac aatctcacta aggctgagag gggcggactg
     3001 agcgaactgg acaaagcagg attcattaaa cggcaacttg tggagactcg gcagattact
     3061 aaacatgtcg cccaaatcct tgactcacgc atgaatacca agtacgacga aaacgacaaa
     3121 cttatccgcg aggtgaaggt gattaccctg aagtccaagc tggtcagcga tttcagaaag
     3181 gactttcaat tctacaaagt gcgggagatc aataactatc atcatgctca tgacgcatat
     3241 ctgaatgccg tggtgggaac cgccctgatc aagaagtacc caaagctgga aagcgagttc
     3301 gtgtacggag actacaaggt ctacgacgtg cgcaagatga ttgccaaatc tgagcaggag
     3361 atcggaaagg ccaccgcaaa gtacttcttc tacagcaaca tcatgaattt cttcaagacc
     3421 gaaatcaccc ttgcaaacgg tgagatccgg aagaggccgc tcatcgagac taatggggag
     3481 actggcgaaa tcgtgtggga caagggcaga gatttcgcta ccgtgcgcaa agtgctttct
     3541 atgcctcaag tgaacatcgt gaagaaaacc gaggtgcaaa ccggaggctt ttctaaggaa
     3601 tcaatcctcc ccaagcgcaa ctccgacaag ctcattgcaa ggaagaagga ttgggaccct
     3661 aagaagtacg gcggattcga ttcaccaact gtggcttatt ctgtcctggt cgtggctaag
     3721 gtggaaaaag gaaagtctaa gaagctcaag agcgtgaagg aactgctggg tatcaccatt
     3781 atggagcgca gctccttcga gaagaaccca attgactttc tcgaagccaa aggttacaag
     3841 gaagtcaaga aggaccttat catcaagctc ccaaagtata gcctgttcga actggagaat
     3901 gggcggaagc ggatgctcgc ctccgctggc gaacttcaga agggtaatga gctggctctc
     3961 ccctccaagt acgtgaattt cctctacctt gcaagccatt acgagaagct gaaggggagc
     4021 cccgaggaca acgagcaaaa gcaactgttt gtggagcagc ataagcatta tctggacgag
     4081 atcattgagc agatttccga gttttctaaa cgcgtcattc tcgctgatgc caacctcgat
     4141 aaagtcctta gcgcatacaa taagcacaga gacaaaccaa ttcgggagca ggctgagaat
     4201 atcatccacc tgttcaccct caccaatctt ggtgcccctg ccgcattcaa gtacttcgac
     4261 accaccatcg accggaaacg ctatacctcc accaaagaag tgctggacgc caccctcatc
     4321 caccagagca tcaccggact ttacgaaact cggattgacc tctcacagct cggaggggat
     4381 taatgaggca tgcaagcttt tatcagaaga actcgtcaag aaggcgatag aaggcgatgc
     4441 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     4501 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     4561 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     4621 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     4681 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcccgatcga
     4741 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     4801 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     4861 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     4921 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     4981 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     5041 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccgggac acggcggcat
     5101 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     5161 ccggagaacc tgcgtgcaat ccatcttgtt caatcataag ggccctcctt catgaaggat
     5221 taatgtcatt atttatttat ggggtatgga gagggatatt tcaaggcgca caaatatgat
     5281 tccaggctcg ttctgcggtt ccgcctgggt gttcgctgag ataggttttt acttcgttat
     5341 acgaaagcgg cgaatcgcca aaaatttttt ttccaagttc gctgcgaaac gaagccataa
     5401 gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
     5461 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
     5521 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
     5581 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     5641 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     5701 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     5761 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     5821 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5881 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5941 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6001 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6061 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6121 atgagcactt ttctcgagtt taagtcacgc ggccgcaaag ttctgctatg tggcgcggta
     6181 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6241 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6301 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6361 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6421 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6481 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6541 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6601 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6661 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6721 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6781 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6841 attgatttct gaaagcgacc aggtgctcgg cgtggcaaga ctcgcagcga acccgtagaa
     6901 agccatgctc cagccgcccg cattggagaa attcttcaaa ttcccgttgc acatagcccg
     6961 gcaattcctt tccctgctct gccataagcg cagcgaatgc cgggtaatac tcgtcaacga
     7021 tctgatagag aagggtttgc tcgggtcggt ggctctggta acgaccagta tcccgatccc
     7081 ggctggccgt cctggccgcc acatgaggca tgttccgcgt ccttgcaata ctgtgtttac
     7141 atacagtcta tcgcttagcg gaaagttctt ttaccctcag ccgaaatgcc tgccgttgct
     7201 agacattgcc agccagtgcc cgtcactccc gtactaactg tcacgaaccc ctgcaataac
     7261 tgtcacgccc ccctgcaata actgtcacga acccctgcaa taactgtcac gcccccaaac
     7321 ctgcaaaccc agcaggggcg ggggctggcg gggtgttgga aaaatccatc catgattatc
     7381 taagaataat ccactaggcg cggttatcag cgcccttgtg gggcgctgct gcccttgccc
     7441 aatatgcccg gccagaggcc ggatagctgg tctattcgct gcgctaggct acacaccgcc
     7501 ccaccgctgc gcggcagggg gaaaggcggg caaagcccgc taaaccccac accaaacccc
     7561 gcagaaatac gctggagcgc ttttagccgc tttagcggcc tttcccccta cccgaagggt
     7621 gggggcgcgt gtgcagcccc gcagggcctg tctcggtcga tcattcagcc cggctcatcc
     7681 ttctggcgtg gcggcagacc gaacaaggcg cggtcgtggt cgcgttcaag gtacgcatcc
     7741 attgccgcca tgagccgatc ctccggccac tcgctgctgt tcaccttggc caaaatcatg
     7801 gcccccacca gcaccttgcg ccttgtttcg ttcttgcgct cttgctgctg ttcccttgcc
     7861 cgcacccgct gaatttcggc attgattcgc gctcgttgtt cttcgagctt ggccagccga
     7921 tccgccgcct tgttgctccc cttaaccatc ttgacacccc attgttaatg tgctgtctcg
     7981 taggctatca tggagccgaa aagcaaaagc aacagcgagg cagcatggcg atttatcacc
     8041 ttacggcgaa aaccggcagc aggtcgggcg gccaatcggc cagggccaag gccgactaca
     8101 tccagcgcga aggcaagtat gcccgcgaca tggatgaagt cttgcacgcc gaatccgggc
     8161 acatgccgga gttcgtcgag cggcccgccg actactggga tgctgccgac ctgtatgaac
     8221 gcgccaatgg gcggctgttc aaggaggtcg aatttgccct gccggtcgag ctgaccctcg
     8281 accagcagaa ggcgctggcg tccgagttcg cccagcacct gaccggtgcc gagcgcctgc
     8341 cgtatacgct ggccatccat gccggtggcg gcgagaaccc gcactgccac ctgatgatct
     8401 ccgagcggat caatgacggc atcgagcggc ccgccgctca gtggttcaag cggtacaacg
     8461 gcaagacccc ggagaagggc ggggcacaga agaccgaagc gctcaagccc aaggcatggc
     8521 ttgagcagac ccgcgaggca tgggccgacc atgccaaccg ggcattagag cgggctggcc
     8581 acgacgcccg cattgaccac agaacacttg aggcgcaggg catcgagcgc ctgcccggtg
     8641 ttcacctggg gccgaacgtg gtggagatgg aaggccgggg catccgcacc gaccgggcag
     8701 acgtggccct gaacatcgac accgccaacg cccagatcat cgacttacag gaataccggg
     8761 aggcaataga ccatgaacgc aatcgacaga gtgaagaaat ccagaggcat caacgagtta
     8821 gcggagcaga tcgaaccgct ggcccagagc atggcgacac tggccgacga agcccggcag
     8881 gtcatgagcc agacccagca ggccagcgag gcgcaggcgg cggagtggct gaaagcccag
     8941 cgccagacag gggcggcatg ggtggagctg gccaaagagt tgcgggaggt agccgccgag
     9001 gtgagcagcg ccgcgcagag cgcccggagc gcgtcgcggg ggtggcactg gaagctatgg
     9061 ctaaccgtga tgctggcttc catgatgcct acggtggtgc tgctgatcgc atcgttgctc
     9121 ttgctcgacc tgacgccact gacaaccgag gacggctcga tctggctgcg cttggtggcc
     9181 cgatgaagaa cgacaggact ttgcaggcca taggccgaca gctcaaggcc atgggctgtg
     9241 agcgcttcga tatcggcgtc agggacgcca ccaccggcca gatgatgaac cgggaatggt
     9301 cagccgccga agtgctccag aacacgccat ggctcaagcg gatgaatgcc cagggcaatg
     9361 acgtgtatat caggcccgcc gagcaggagc ggcatggtct ggtgctggtg gacgacctca
     9421 gcgagtttga cctggatgac atgaaagccg agggccggga gcctgccctg gtagtggaaa
     9481 ccagcccgaa gaactatcag gcatgggtca aggtggccga cgccgcaggc ggtgaacttc
     9541 gggggcagat tgcccggacg ctggccagcg agtacgacgc cgacccggcc agcgccgaca
     9601 gccgccacta tggccgcttg gcgggcttca ccaaccgcaa ggacaagcac accacccgcg
     9661 ccggttatca gccgtgggtg ctgctgcgtg aatccaaggg caagaccgcc accgctggcc
     9721 cggcgctggt gcagcaggct ggccagcaga tcgagcaggc ccagcggcag caggagaagg
     9781 cccgcaggct ggccagcctc gaactgcccg agcggcagct tagccgccac cggcgcacgg
     9841 cgctggacga gtaccgcagc gagatggccg ggctggtcaa gcgcttcggt gatgacctca
     9901 gcaagtgcga ctttatcgcc gcgcagaagc tggccagccg gggccgcagt gccgaggaaa
     9961 tcggcaaggc catggccgag gccagcccag cgctggcaga gcgcaagccc ggccacgaag
    10021 cggattacat cgagcgcacc gtcagcaagg tcatgggtct gcccagcgtc cagcttgcgc
    10081 gggccgagct ggcacgggca ccggcacccc gccagcgagg catggacagg ggcgggccag
    10141 atttcagcat gtagtgcttg cgttggtact cacgcctgtt atactatgag tactcacgca
    10201 cagaaggggg ttttatggaa tacgaaaaaa gcgcttcagg gtcggtctac ctgatcaaaa
    10261 gtgacaaggg ctattggttg cccggtggct ttggttatac gtcaaacaag gccgaggctg
    10321 gccgcttttc agtcgctgat atggccagcc ttaaccttga cggctgcacc ttgtccttgt
    10381 tccgcgaaga caagcctttc ggccccggca agtttctcgg tgactgatat gaaagaccaa
    10441 aaggacaagc agaccggcga cctgctggcc agccctgacg ctgtacgcca agcgcgatat
    10501 gccgagcgca tgaaggccaa agggatgcgt cagcgcaagt tctggctgac cgacgacgaa
    10561 tacgaggcgc tgcgcgagtg cctggaagaa ctcagagcgg cgcagggcgg gggtagtgac
    10621 cccgccagcg cctaaccacc aactgcctgc aaaggaggca atcaatggct acccataagc
    10681 ctatcaatat tctggaggcg ttcgcagcag cgccgccacc gctggactac gttttgccca
    10741 acatggtggc cggtacggtc ggggcgctgg tgtcgcccgg tggtgccggt aaatccatgc
    10801 tggccctgca actggccgca cagattgcag gcgggccgga tctgctggag gtgggcgaac
    10861 tgcccaccgg cccggtgatc tacctgcccg ccgaagaccc gcccaccgcc attcatcacc
    10921 gcctgcacgc ccttggggcg cacctcagcg ccgaggaacg gcaagccgtg gctgacggcc
    10981 tgctgatcca gccgctgatc ggcagcctgc ccaacatcat ggccccggag tggttcgacg
    11041 gcctcaagcg cgccgccgag ggccgccgcc tgatggtgct ggacacgctg cgccggttcc
    11101 acatcgagga agaaaacgcc agcggcccca tggcccaggt catcggtcgc atggaggcca
    11161 tcgccgccga taccgggtgc tctatcgtgt tcctgcacca tgccagcaag ggcgcggcca
    11221 tgatgggcgc aggcgaccag cagcaggcca gccggggcag ctcggtactg gtcgataaca
    11281 tccgctggca gtcctacctg tcgagcatga ccagcgccga ggccgaggaa tggggtgtgg
    11341 acgacgacca gcgccggttc ttcgtccgct tcggtgtgag caaggccaac tatggcgcac
    11401 cgttcgctga tcggtggttc aggcggcatg acggcggggt gctcaagccc gccgtgctgg
    11461 agaggcagcg caagagcaag ggggtgcccc gtggtgaagc ctaagaacaa gcacagcctc
    11521 agccacgtcc ggcacgaccc ggcgcactgt ctggcccccg gcctgttccg tgccctcaag
    11581 cggggcgagc gcaagcgcag caagctggac gtgacgtatg actacggcga cggcaagcgg
    11641 atcgagttca gcggcccgga gccgctgggc gctgatgatc tgcgcatcct gcaagggctg
    11701 gtggccatgg ctgggcctaa tggcctagtg cttggcccgg aacccaagac cgaaggcgga
    11761 cggcagctcc ggctgttcct ggaacccaag tgggaggccg tcaccgctga tgccatggtg
    11821 gtcaaaggta gctatcgggc gctggcaaag gaaatcgggg cagaggtcga tagtggtggg
    11881 gcgctcaagc acatacagga ctgcatcgag cgcctttgga aggtatccat catcgcccag
    11941 aatggccgca agcggcaggg gtttcggctg ctgtcggagt acgccagcga cgaggcggac
    12001 gggcgcctgt acgtggccct gaaccccttg atcgcgcagg ccgtcatggg tggcggccag
    12061 catgtgcgca tcagcatgga cgaggtgcgg gcgctggaca gcgaaaccgc ccgcctgctg
    12121 caccagcggc tgtgtggctg gatcgacccc ggcaaaaccg gcaaggcttc catagatacc
    12181 ttgtgcggct atgtctggcc gtcagaggcc agtggttcga ccatgcgcaa gcgccgccag
    12241 cgggtgcgcg aggcgttgcc ggagctggtc gcgctgggct ggacggtaac cgagttcgcg
    12301 gcgggcaagt acgacatcac ccggcccaag gcggcaggct gacccccccc actctattgt
    12361 aaacaagaca tttttatctt ttatattcaa tggcttattt tcctgctaat tggtaatacc
    12421 atgaaaaata ccatgctcag aaaaggctta acaatatttt gaaaaattgc ctactgagcg
    12481 ctgccgcaca gctccatagg ccgctttcct ggctttgctt ccagatgtat gctcttctgc
    12541 tcccgaacgc cagcaagacg tagcccagcg cgtcggccag cttgcaattc gcgctaactt
    12601 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    12661 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
    12721 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
    12781 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
    12841 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
    12901 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
    12961 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
    13021 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
    13081 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
    13141 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
    13201 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
    13261 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
    13321 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
    13381 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
    13441 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
    13501 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
    13561 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
    13621 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
    13681 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
    13741 tgccataccg cgaaaggttt tgcaccattc gatggtgtca acgtaaatgc cgcttcgcct
    13801 tcgcgcgcga attgcaagct gatccgggct tatcgactgc acggtgcacc aatgcttctg
    13861 gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat
    13921 tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg
    13981 gttctggcaa atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt
    14041 ggaattgtga gcggataaca atttcacaca ggaaaca
//
