LOCUS       40924_13350        3999 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pCRB26 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3999)
  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 3999)
  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 3999)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3999)
  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..3999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    202..223
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        238..268
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    276..292
                     /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     300..316
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    326..382
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(386..402)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(728..1316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1365..2815
                     /note="derived from Corynebacteriumt glutamicum cryptric
                     plasmid pCASE1"
     CDS             complement(1421..1888)
                     /codon_start=1
                     /gene="rep2"
                     /product="replicase"
                     /label=rep2
                     /protein_id="BAI66096.1"
                     /translation="MNHYLWPKLTVLPGQSINGSSPSHACGQTAPPSTTPHSPQCNPHA
                     GRKAGNEALRCVVRLDILFGGTLAKVYARYPRNGKTARELAERAGMSVRTAQRWTSEPR
                     EVFIKRANEKRARVQELRAKGLSMRAIAAEIGCSVGTVHRYVKEVEEKKTA"
     gene            complement(1421..1888)
                     /gene="rep2"
                     /label=rep2
     CDS             complement(1680..2615)
                     /codon_start=1
                     /gene="rep1"
                     /product="replicase"
                     /label=rep1
                     /protein_id="BAI66097.1"
                     /translation="MKTDRARSCEKVSYMRPTTRPEGRFEQLWLPLWPLASDDLREGIY
                     RTSRKNALDKRYVEANPDALSNLLVVDIDQEDALLRSLWDREDWRPNAVVENPLNGHAH
                     AVWALAEPFTRTEYAKRKPLAYAAAVTEGLRRSVDGDSGYSGLITKNPEHTAWDSHWIT
                     DKLYTLDELRFWLEETGFMPPASWRKTRRFSPVGLGRNCALFESARTWAYREVRKHFGD
                     ADGLGRAIQTTAQALNQELFDEPLPVAEVDCIARSIHKWIITKSRMWTDGAAVYDATFT
                     AMQSARGKKGWQRSAEVRREAGHTLWRNIG"
     gene            complement(1680..2615)
                     /gene="rep1"
                     /label=rep1
     CDS             complement(2852..3874)
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     protein_bind    complement(3923..3939)
                     /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)."
     promoter        complement(3947..3977)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 aaggcagtta ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc
       61 ggatgaatgg cagaaattca gcttggccca gtgccaagct ccaatacgca aaccgcctct
      121 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      181 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      241 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      301 aggaaacagc tatgaccatg attacgaatt cgagctcggt acccggggat cctctagagt
      361 cgacctgcag gcatgcaagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      421 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
      481 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
      541 gagcttcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      601 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      661 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      721 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      781 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      841 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      901 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      961 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1021 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1081 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1141 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1201 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1261 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1321 atcctttgat cttttctacg gggtctgacg ctcagtggaa ctagatctga cttggttacg
     1381 atggactttg aacacgccga gggtgactaa accgctggat ttacgcggtt ttcttctctt
     1441 caacttcttt gacgtagcgg tgaaccgtgc ccaccgagca accaatctct gccgcgatag
     1501 cgcgcatgga cagacctttg gcgcgcagct cctggacgcg agcacgcttc tcgttggcac
     1561 gtttaatgaa cacttcacgc ggttcggaag tccatcgttg agctgttctc accgacatac
     1621 cggcacgttc tgccagctcg cgggctgttt ttccgttgcg tggataacgt gcataaacct
     1681 tagccaatgt tcctccaaag agtatgtcca gcctcacgac gcacctcagc gcttcgttgc
     1741 cagcctttct tcccgcgtgc ggattgcatt gcggtgaatg tggcgtcgta gacggcggcg
     1801 ccgtctgtcc acatgcgtga cttggtgatg atccatttat ggattgacct ggcaatacag
     1861 tcaacttcgg ccacaggtag tggttcatca aacagctctt ggttaagtgc ttgcgcggtg
     1921 gtttggattg cgcggcctag gccgtcagcg tctccaaaat gctttctgac ctcccgatat
     1981 gcccacgtac gtgcgctttc aaagagtgcg caattacgac ctagaccaac tggcgagaac
     2041 cgccgcgttt tcctccagga cgcaggcggc ataaagccgg tttcttcgag ccaaaagcgc
     2101 agctcatcga gcgtatacag cttatcggtg atccagtgac tatcccatgc agtgtgctcg
     2161 gggtttttgg tgatcagccc ggagtatccg ctatcgccat cgacagagcg ccgtaggcct
     2221 tcggtgacag ccgcggcata ggccaaaggc ttgcgtttgg cgtattcggt gcgggtaaat
     2281 ggctccgcga gcgcccagac agcgtgtgcg tgcccgttta aggggttttc aaccaccgcg
     2341 ttaggtctcc agtcctccct gtcccacaaa gagcgcaaaa gcgcgtcctc ctggtcgatg
     2401 tcaacgacca ggaggttaga gagcgcgtcg ggattggctt cgacgtagcg cttatccagc
     2461 gcgttcttcc gtgaggtgcg gtaaatgccc tcacggaggt catcgcttgc caatggccac
     2521 agcggcagcc acagctgctc aaagcgtccc tcagggcggg tagttggtct catgtagctg
     2581 actttctcac acgagcgtgc acggtcggtt ttcattcctc ggcgtgttca taatacgaca
     2641 tttaaccaag tcagatgttt ccccggtttc cgggggttcc cctgaagaac ccttccagtg
     2701 cgagcgaagc gagctccttt ggccggcgcc cctcaggtag ccctctaagg ctcccagggc
     2761 tccgcccctc cctgaggttg gctcaagcct cctggtggct cctacggacg ttctagatct
     2821 gtcgaacgga agatcacttc ggatatctct attcctttgc cctcggacga gtgctggggc
     2881 gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc
     2941 tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc
     3001 gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt
     3061 caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccggatgcc
     3121 tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga
     3181 tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg
     3241 ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc
     3301 ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg
     3361 cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc
     3421 atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc
     3481 cgctcgtctg gctaagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca
     3541 gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg
     3601 agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga
     3661 gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc
     3721 tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt
     3781 cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct
     3841 cgacagacgt cgcggtgagt tcaggctttt tcatatcggg gtcgtcctct ccaaatgaac
     3901 tcgagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3961 gaagcataaa gtgtaaagcc tggggtgcct aatgatatc
//
