LOCUS       V009319                 9274 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009319
VERSION     V009319
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9274)
  AUTHORS   Sato'o Y, Hisatsune J, Sakuma T, Yamamoto T, Sugai M.
  TITLE     Novel CRISPR interference technology, an elementary method for
            inspects of Staphylococcus aureus clinical isolates
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9274)
  AUTHORS   Sato'o Y, Hisatsune J, Sugai M.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-FEB-2016) Contact:Motoyuki Sugai Hiroshima University,
            Bacteriology; Minamiku, Kasumi1-2-3, Hiroshima, Hiroshima 734-8551,
            Japan URL :http://home.hiroshima-u.ac.jp/saikin/index.html
REFERENCE   3  (bases 1 to 9274)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9274)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: CLC Genomics Workbench v. 7
            Sequencing Technology :: Illumina MiSeq
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (22-FEB-2016) Contact:Motoyuki Sugai Hiroshima University,
            Bacteriology"; volume: " Minamiku, Kasumi1-2-3, Hiroshima, Hiroshima
            734-8551, Japan URL :http"; pages:
            "//home.hiroshima-u.ac.jp/saikin/index.htm"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        37..67
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /label="lac operator"
                     /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     99..115
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     repeat_region   414..449
                     /rpt_unit_seq="gttttagagctatgctgttttgaatggtcccaaaac"
     misc_feature    complement(450..455)
                     /label="BsaI recognition site"
                     /note="BsaI recognition site"
     misc_feature    473..478
                     /label="BsaI recognition site"
                     /note="BsaI recognition site"
     repeat_region   480..515
                     /label="DR"
                     /note="direct repeat for the Streptococcus pyogenes
                     CRISPR/Cas system"
     misc_feature    complement(516..647)
                     /label="crRNA leader"
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     CDS             complement(671..4774)
                     /label="dCas9"
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     misc_RNA        5070..5148
                     /label="tracrRNA"
                     /note="trans-activating CRISPR RNA for the Streptococcus
                     pyogenes CRISPR/Cas9 system"
     primer_bind     complement(5456..5472)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      5823..6411
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6988..7635)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             complement(7917..8918)
                     /label="repB"
                     /note="RepB replication protein"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cttgcatgcc tgcagcgatg gatatgttct gccaaggaat tgcatcaacg
      181 catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatcacac
      241 tactcttctt ttgcctatta taacatgaaa acgctttaac agaaagaata ggaaggtatc
      301 cgactgctgg tattaaccct ctttctcaag ttatcatcgg caatgttgaa tggagtccat
      361 tcaaaacagc atagctctaa aacatggaac tcaacaagtc tcagtgtgct gaagttttgg
      421 gaccattcaa aacagcatag ctctaaaacg agacctttga gcttccgaga ctggtctcag
      481 ttttgggacc attcaaaaca gcatagctct aaaacctcgt agactatttt tgtctaaaaa
      541 atttcgtaat cgcactattt gtctcagcta gacttcagtc ttgaaaagcc cctgtattac
      601 tgcatttatt aagagtatta taccatattt ttagttatta agaaataatc ttcatctaaa
      661 atatacttca gtcacctcct agctgactca aatcaatgcg tgtttcataa agaccagtga
      721 tggattgatg gataagagtg gcatctaaaa cttcttttgt agacgtatat cgtttacgat
      781 caattgttgt atcaaaatat ttaaaagcag cgggagctcc aagattcgtc aacgtaaata
      841 aatgaataat attttctgct tgttcacgta ttggtttgtc tctatgtttg ttatatgcac
      901 taagaacttt atctaaattg gcatctgcta aaataacacg cttagaaaat tcactgattt
      961 gctcaataat ctcatctaaa taatgcttat gctgctccac aaacaattgt ttttgttcgt
     1021 tatcttctgg actacccttc aacttttcat aatgactagc taaatataaa aaattcacat
     1081 atttgcttgg cagagccagc tcatttcctt tttgtaattc tccggcacta gccagcatcc
     1141 gtttacgacc gttttttaac tcaaaaagac tatatttagg tagtttaatg attaagtctt
     1201 ttttaacttc cttatatcct ttagcttcta aaaagtcaat cggatttttt tcaaaggaac
     1261 ttctttccat aattgtgatc cctagtaact ctttaacgga ttttaacttc ttcgatttcc
     1321 ctttttccac cttagcaacc actaggactg aataagctac cgttggacta tcaaaaccac
     1381 catatttttt tggatcccag tcttttttac gagcaataag cttgtccgaa tttctttttg
     1441 gtaaaattga ctccttggag aatccgcctg tctgtacttc tgttttcttg acaatattga
     1501 cttggggcat ggacaatact ttgcgcactg tggcaaaatc tcgcccttta tcccagacaa
     1561 tttctccagt ttccccatta gtttcgatta gagggcgttt gcgaatctct ccatttgcaa
     1621 gtgtaatttc tgttttgaag aagttcatga tattagagta aaagaaatat tttgcggttg
     1681 ctttgcctat ttcttgctca gacttagcaa tcattttacg aacatcataa actttataat
     1741 caccatagac aaactccgat tcaagttttg gatatttctt aatcaaagca gttccaacga
     1801 cggcatttag atacgcatca tgggcatgat ggtaattgtt aatctcacgt actttataga
     1861 attggaaatc ttttcggaag tcagaaacta atttagattt taaggtaatc actttaacct
     1921 ctcgaataag tttatcattt tcatcgtatt tagtattcat gcgactatcc aaaatttgtg
     1981 ccacatgctt agtgatttgg cgagtttcaa ccaattggcg tttgataaaa ccagctttat
     2041 caagttcact caaacctcca cgttcagctt tcgttaaatt atcaaactta cgttgagtga
     2101 ttaacttggc gtttagaagt tgtctccaat agtttttcat ctttttgact acttcttcac
     2161 ttggaacgtt atccgattta ccacgatttt tatcagaacg cgttaagacc ttattgtcta
     2221 ttgaatcgtc tttaaggaaa ctttgtggaa caattgcatc gacatcataa tcacttaaac
     2281 gattaatatc taattcttgg tccacataca tgtctcttcc attttggaga taatagagat
     2341 agagcttttc attttgcaat tgagtatttt caacaggatg ctctttaaga atctgacttc
     2401 ctaattcttt gataccttct tcgattcgtt tcatacgctc tcgcgaattt ttctggccct
     2461 tttgagttgt ctgattttca cgtgccattt caataacgat attttctggc ttatgccgcc
     2521 ccattacttt gaccaattca tcaacaactt ttacagtctg taaaatacct tttttaatag
     2581 cagggctacc agctaaattt gcaatatgtt catgtaaact atcgccttgt ccagacactt
     2641 gtgctttttg aatgtcttct ttaaatgtca aactatcatc atggatcagc tgcataaaat
     2701 tgcgattggc aaaaccatct gatttcaaaa aatctaatat tgttttgcca gattgcttat
     2761 ccctaatacc attaatcaat tttcgagaca aacgtcccca accagtataa cggcgacgtt
     2821 taagctgttt catcacctta tcatcaaaga ggtgagcata tgttttaagt ctttcctcaa
     2881 tcatctccct atcttcaaat aaggtcaatg ttaaaacaat atcctctaag atatcttcat
     2941 tttcttcatt atccaaaaaa tctttatctt taataatttt tagcaaatca tggtaggtac
     3001 ctaatgaagc attaaatcta tcttcgactc ctgaaatttc aacactatca aaacattcta
     3061 tttttttgaa ataatcttct tttaattgct taacggttac ttttcgattt gttttgaaga
     3121 gtaaatcaac aatggctttc ttctgttcac ctgaaagaaa tgctggtttt cgcattcctt
     3181 cagtaacata tttgaccttt gtcaattcgt tataaaccgt aaaatactca taaagcaaac
     3241 tatgttttgg tagtactttt tcatttggaa gatttttatc aaagtttgtc atgcgttcaa
     3301 taaatgattg agctgaagca cctttatcga caacttcttc aaaattccat ggggtaattg
     3361 tttcttcaga cttccgagtc atccatgcaa aacgactatt gccacgcgcc aatggaccaa
     3421 cataataagg aattcgaaaa gtcaagattt tttcaatctt ctcacgattg tcttttaaaa
     3481 atggataaaa gtcttcttgt cttctcaaaa tagcatgcag ctcacccaag tgaatttgat
     3541 ggggaataga gccgttgtca aaggtccgtt gcttgcgcag caaatcttca cgatttagtt
     3601 tcaccaataa ttcctcagta ccatccattt tttctaaaat tggtttgata aatttataaa
     3661 attcttcttg gctagctccc ccatcaatat aacctgcata tccgtttttt gattgatcaa
     3721 aaaagatttc tttatacttt tctggaagtt gttgtcgaac taaagctttt aaaagagtca
     3781 agtcttgatg atgttcatcg tagcgtttaa tcattgaagc tgatagggga gccttagtta
     3841 tttcagtatt tactcttagg atatctgaaa gtaaaatagc atctgataaa ttcttagctg
     3901 ccaaaaacaa atcagcatat tgatctccaa tttgcgccaa taaattatct aaatcatcat
     3961 cgtaagtatc ttttgaaagc tgtaatttag catcttctgc caaatcaaaa tttgatttaa
     4021 aattaggggt caaacccaat gacaaagcaa tgagattccc aaataagcca tttttcttct
     4081 caccggggag ctgagcaatg agattttcta atcgtcttga tttactcaat cgtgcagaaa
     4141 gaatcgcttt agcatctact ccacttgcgt taatagggtt ttcttcaaat aattgattgt
     4201 aggtttgtac caactggata aatagtttgt ccacatcact attatcagga tttaaatctc
     4261 cctcaatcaa aaaatgacca cgaaacttaa tcatatgcgc taaggccaaa tagattaagc
     4321 gcaaatccgc tttatcagta gaatctacca atttttttcg cagatgatag atagttggat
     4381 atttctcatg ataagcaact tcatctacta tatttccaaa aataggatga cgttcatgct
     4441 tcttgtcttc ttccaccaaa aaagactctt caagtcgatg aaagaaacta tcatctactt
     4501 tcgccatctc atttgaaaaa atctcctgta gataacaaat acgattcttc cgacgtgtat
     4561 accttctacg agctgtccgt ttgagacgag tcgcttccgc tgtctctcca ctgtcaaata
     4621 aaagagcccc tataagattt tttttgatac tgtggcggtc tgtatttccc agaaccttga
     4681 actttttaga cggaacctta tattcatcag tgatcaccgc ccatccgacg ctatttgtgc
     4741 cgatagctaa gcctattgag tatttcttat ccatttttgc ctcctaaaat aaaaagttta
     4801 aattaaatcc ataatgagtt tgatgatttc aataatagtt ttaatgacct ccgaaattag
     4861 tttaatatgc tttaattttt ctttttcaaa atatctcttc aaaaaatatt acccaatact
     4921 taataataaa tagattataa cacaaaattc ttttaaaaag tagtttattt tgttatcatt
     4981 ctatagtatt aagtattgtt ttatggctga taaatttctt tgaatttctc cttgattatt
     5041 tgttataaaa gttataaaat aatcttgttg gaaccattca aaacagcata gcaagttaaa
     5101 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttgatac
     5161 ttctattcta ctctgactgc aaaccaaaaa aacaagcgct ttcaaaacgc ttgttttatc
     5221 atttttaggg aaattaatct cttaatcctt ttatcattct acatttaggc gctgccatct
     5281 tgctaaacct actaagctcc acaggatgat ttcgtaatcc cgcaagaggc ccggcagtac
     5341 cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag
     5401 cgcattgtta ctgcaggtcg actctagagg atccccgggt accgagctcg aattcactgg
     5461 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     5521 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     5581 cccaacagtt gcgcagcctg aatggcgaat gagcttggca ctggcggtgc ctcactgatt
     5641 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5701 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5761 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5821 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5881 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5941 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6001 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6061 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6121 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6181 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6241 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6301 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6361 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6421 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcaggc aaccacttcc
     6481 aagtaaagta taacacacta tactttatat tcataaagtg tgtgctctgc gaggctgtcg
     6541 gcagtgccga ccaaaaccat aaaaccttta agacctttct tttttttacg agaaaaaaga
     6601 aacaaaaaaa cctgccctct gccacctcag caaagggggg ttttgctctc gtgctcgttt
     6661 aaaaatcagc aagggacagg tagtattttt tgagaagatc actcaaaaaa tctccacctt
     6721 taaacccttg ccaattttta ttttgtccgt tttgtctagc ttaccgaaag ccagactcag
     6781 caagaataaa atttttattg tctttcggtt ttctagtgta acggacaaaa ccactcaaaa
     6841 taaaaaagat acaagagagg tcactcgtac ttttattcag caatcggatc tggagctgta
     6901 atataaaaac cttcttcaac taacggggca ggttagtgac attagaaaac cgactgtaaa
     6961 aagtacagtc ggcattatct catattataa aagccagtca ttaggcctat ctgacaattc
     7021 ctgaatagag ttcataaaca atcctgcatg ataaccatca caaacagaat gatgtacctg
     7081 taaagatagc ggtaaatata ttgaattacc tttattaatg aattttcctg ctgtaataat
     7141 gggtagaagg taattactat tattattgat atttaagtta aacccagtaa atgaagtcca
     7201 tggaataata gaaagagaaa aagcattttc aggtataggt gttttgggaa acaatttccc
     7261 cgaaccatta tatttctcta catcagaaag gtataaatca taaaactctt tgaagtcatt
     7321 ctttacagga gtccaaatac cagagaatgt tttagataca ccatcaaaaa ttgtataaag
     7381 tggctctaac ttatcccaat aacctaactc tccgtcgcta ttgtaaccag ttctaaaagc
     7441 tgtatttgag tttatcaccc ttgtcactaa gaaaataaat gcagggtaaa atttatatcc
     7501 ttcttgtttt atgtttcggt ataaaacact aatatcaatt tctgtggtta tactaaaagt
     7561 cgtttgttgg ttcaaataat gattaaatat ctcttttctc ttccaattgt ctaaatcaat
     7621 tttattaaag ttcatttgat atgcctccta aatttttatc taaagtgaat ttaggaggct
     7681 tacttgtctg ctttcttcat tagaatcaat ccttttttaa aagtcaatgc tcgtcgtata
     7741 acagatgcga tgatgcagac caatcaacat ggcacctgcc attgctacct gtacagtcaa
     7801 ggatggtaga aatgttgtcg gtcctattaa taactcgtct tcctaagcat ccttcaatcc
     7861 ttttaataac aattatagca tctaatcttc aacaaactgg cccgtttgtt gaactactct
     7921 ttaataaaat aatttttccg ttcccaattc cacattgcaa taatagaaaa tccatcttca
     7981 tcggcttttt cgtcatcatc tgtatgaatc aaatcgcctt cttctgtgtc atcaaggttt
     8041 aattttttat gtatttcttt taacaaacca ccataggaga ttaacctttt acggtgtaaa
     8101 ccttcctcca aatcagacaa acgtttcaaa ttcttttctt catcatcggt cataaaatcc
     8161 gtatccttta caggatattt tgcagtttcg tcaattgccg attgtatatc cgatttatat
     8221 ttatttttcg gtcgaatcat ttgaactttt acatttggat catagtctaa tttcattgcc
     8281 tttttccaaa attgaatcca ttgtttttga ttcacgtagt tttctgtatt cttaaaataa
     8341 gttggttcca cacataccaa tacatgcatg tgctgattat aagaattatc tttattattt
     8401 attgtcactt ccgttgcacg cataaaacca acaagatttt tattaatttt tttatattgc
     8461 atcattcggc gaaatccttg agccatatct gacaaactct tatttaattc ttcgccatca
     8521 taaacatttt taactgttaa tgtgagaaac aaccaacgaa ctgttggctt ttgtttaata
     8581 acttcagcaa caaccttttg tgactgaatg ccatgtttca ttgctctcct ccagttgcac
     8641 attggacaaa gcctggattt acaaaaccac actcgataca actttctttc gcctgtttca
     8701 cgattttgtt tatactctaa tatttcagca caatctttta ctctttcagc ctttttaaat
     8761 tcaagaatat gcagaagttc aaagtaatca acattagcga ttttcttttc tctccatggt
     8821 ctaacttttc cactttttgt cttgtccact aaaacccttg atttttcatc tgaataaatg
     8881 ctactattag gacacataat attaaaagaa acccccatct atttagttat ttgtttagtc
     8941 acttataact ttaacagatg gggtttttct gtgcaaccaa ttttaagggt tttcaatact
     9001 ttaaaacaca tacataccaa cacttcaacg cacctttcag caactaaaat aaaaatgacg
     9061 ttatttctat atgtatcaag ataagaaaga acaagttcaa aaccatcaaa aaaagacacc
     9121 ttttcaggtg ctttttttat tttataaact cattccctga tctcgacttc gttctttttt
     9181 tacctctcgg ttatgagtta gttcaaattc gttcttttta ggttctaaat cgtgtttttc
     9241 ttggaattgt gctgttttat cctttacctt gtct
//
