LOCUS       40924_13345        4497 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pCRB2 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4497)
  AUTHORS   Tsuchida Y, Kimura S, Suzuki N, Inui M, Yukawa H.
  TITLE     Characterization of a 24-kb plasmid pCGR2 newly isolated from 
            Corynebacterium glutamicum
  JOURNAL   Appl. Microbiol. Biotechnol. 87 (5), 1855-1866 (2010)
  PUBMED    20552356
REFERENCE   2  (bases 1 to 4497)
  AUTHORS   Gunji W, Inui M, Suzuki N, Yukawa H.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-OCT-2009) Contact:Nobuaki Suzuki Research Institute of
            Innovative Technology for the Earth, Molecular Microbiology and 
            Biotechnology Group; 9-2 kizugawadai, Kizugawa, kyoto 619-0292, 
            Japan URL :http://www.rite.or.jp/
REFERENCE   3  (bases 1 to 4497)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4497)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol."; date: "2010"; volume: "87"; issue: "5"; 
            pages: "1855-1866"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-OCT-2009) Contact:Nobuaki Suzuki Research Institute of 
            Innovative Technology for the Earth, Molecular Microbiology and 
            Biotechnology Group; 9-2 kizugawadai, Kizugawa, kyoto 619-0292, 
            Japan URL :http://www.rite.or.jp/"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    6..1818
                     /note="derived from Brevibacterium lactofermentum cryptric 
                     plasmid pBL1"
     CDS             529..1734
                     /codon_start=1
                     /gene="rep"
                     /product="replicase"
                     /label=rep
                     /protein_id="BAI66092.1"
                     /translation="MYKITNSKALAGCRRWRRDEAVAVSWSSNGASQFEGLQNSHSRWG
                     SPLAELEVMGERRIELAIATKNHLAAGGALMMFVGTVRHNRSQSFAQVEAGIKTAYSSM
                     VKTSQWKKERARYGVEHTYSDYEVTDSWANGWHLHRNMLLFLDRPLSDDELKAFEDSMF
                     SRWSAGVVKAGMDAPLREHGVKLDQVPTWGGDAAKMATYLAKGMSQELTGSATKTASKG
                     SYTPFQMLDMLADQSDAGEDMDAVLVARWREYEVGSKNLRSSWSRGAKRALGIDYIDAD
                     VRREMEEELYKLAGLEAPERVESTRVAVALVKPDDWKLIQSDFAVRQYVLDCVDKAKDV
                     AAAQRVANEVLASLGVDSTPCMIVMDDVDLDAVLPTHGDATKRDLNAAVFAGNEQTILR
                     TH"
     gene            529..1734
                     /gene="rep"
                     /label=rep
     protein_bind    1866..1887
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1902..1932
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1940..1956
                     /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     1964..1980
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1990..2046
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2050..2066)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2288..3100)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      3480..4068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcgagacatg cagtcatgtc gtgctaatgt gtaaaacatg tacatgcaga ttgctggggg
       61 tgcagggggc ggagccaccc tgtccatgcg gggtgtgggg cttgccccgc cggtacagac
      121 agtgagcacc ggggcaccta gtcgcggata ccccccctag gtatcggaca cgtaaccctc
      181 ccatgtcgat gcaaatcttt aacattgagt acgggtaagc tggcacgcat agccaagcta
      241 ggcggccacc aaacaccact aaaaattaat agtccctaga caagacaaac ccccgtgcga
      301 gctaccaact catatgcacg ggggccacat aacccgaagg ggtttcaatt gacaaccata
      361 gcactagcta agacaacggg cacaacaccc gcacaaactc gcactgcgca accccgcaca
      421 acatcgggtc taggtaacac tgaaatagaa gtgaacacct ctaaggaacc gcaggtcaat
      481 gagggttcta aggtcactcg cgctagggcg tggcgtaggc aaaacgtcat gtacaagatc
      541 accaatagta aggctctggc ggggtgccgt aggtggcgca gggacgaagc tgttgcggtg
      601 tcctggtcgt ctaacggtgc ttcgcagttt gagggtctgc aaaactctca ctctcgctgg
      661 gggtcacctc tggctgaatt ggaagtcatg ggcgaacgcc gcattgagct ggctattgct
      721 actaagaatc acttggcggc gggtggcgcg ctcatgatgt ttgtgggcac tgttcgacac
      781 aaccgctcac agtcatttgc gcaggttgaa gcgggtatta agactgcgta ctcttcgatg
      841 gtgaaaacat ctcagtggaa gaaagaacgt gcacggtacg gggtggagca cacctatagt
      901 gactatgagg tcacagactc ttgggcgaac ggttggcact tgcaccgcaa catgctgttg
      961 ttcttggatc gtccactgtc tgacgatgaa ctcaaggcgt ttgaggattc catgttttcc
     1021 cgctggtctg ctggtgtggt taaggccggt atggacgcgc cactgcgtga gcacggggtc
     1081 aaacttgatc aggtgcctac ctggggtgga gacgctgcga aaatggcaac ctacctcgct
     1141 aagggcatgt ctcaggaact gactggctcc gctactaaaa ccgcgtctaa ggggtcgtac
     1201 acgccgtttc agatgttgga tatgttggcc gatcaaagcg acgccggcga ggatatggac
     1261 gctgttttgg tggctcggtg gcgtgagtat gaggttggtt ctaaaaacct gcgttcgtcc
     1321 tggtcacgtg gggctaagcg tgctttgggc attgattaca tagacgctga tgtacgtcgt
     1381 gaaatggaag aagaactgta caagctcgcc ggtctggaag caccggaacg ggtcgaatca
     1441 acccgcgttg ctgttgcttt ggtgaagccc gatgattgga aactgattca gtctgatttc
     1501 gcggttaggc agtacgttct agattgcgtg gataaggcta aggacgtggc cgctgcgcaa
     1561 cgtgtcgcta atgaggtgct ggcaagtctg ggtgtggatt ccaccccgtg catgatcgtt
     1621 atggatgatg tggacttgga cgcggttctg cctactcatg gggacgccac taagcgtgat
     1681 ctgaatgcgg cggtgttcgc gggtaatgag cagactattc ttcgcaccca ctaaaagcgg
     1741 cataaacccc gttcgatatt ttgtacgatg aatttatggt caatgtcgcg ggggcaaact
     1801 atgatgggtc ttgttgttag atctaggttt cccgactgga aagcgggcag tgagcgcaac
     1861 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     1921 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     1981 catgattacg aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc
     2041 aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     2101 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     2161 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgcgatt tattcaacaa
     2221 agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     2281 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     2341 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     2401 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     2461 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     2521 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     2581 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     2641 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     2701 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     2761 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     2821 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     2881 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     2941 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     3001 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     3061 gcggcttcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     3121 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     3181 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     3241 aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca
     3301 aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc ctcactttct
     3361 ggctggatga tggggcgatt caggcctggt atgagtcagc aacaccttct tcacgaggca
     3421 gacctctcga cggagttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3481 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3541 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3601 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3661 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3721 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3781 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3841 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3901 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3961 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4021 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4081 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4141 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4201 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga agctcgcaca
     4261 ttcagcagcg tttttcagcg cgttttcgat caacgtttca atgttggtat caacaccagg
     4321 tttaactttg aacttatcgg cactgacggt tactgatttt gaacttttgc tttgccacgg
     4381 aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca aaagttcgcc
     4441 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgc
//
