LOCUS       40924_13330        4569 bp DNA     circular SYN 17-DEC-2018
DEFINITION  C. glutamicum-E. coli shuttle vector pCRB12, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4569)
  AUTHORS   Okibe N, Suzuki N, Inui M, Yukawa H.
  TITLE     Antisense-RNA-mediated plasmid copy number control in pCG1-family 
            plasmids, pCGR2 and pCG1, in Corynebacterium glutamicum
  JOURNAL   Microbiology (Reading, Engl.) 156 (PT 12), 3609-3623 (2010)
  PUBMED    20798162
REFERENCE   2  (bases 1 to 4569)
  AUTHORS   Noburyu R, Inui M.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-APR-2010) Molecular Microbiology and Biotechnology 
            Group, Research Institute of Innovative Technology for the Earth 
            (RITE), 9-2 Kizugawadai, Kizugawa, Kyoto 619-0292, Japan
REFERENCE   3  (bases 1 to 4569)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4569)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Microbiology (Reading, Engl.) 156 (PT 12), 3609-3623 (2010)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-APR-2010) Molecular Microbiology and Biotechnology Group, 
            Research Institute of Innovative Technology for the Earth (RITE), 
            9-2 Kizugawadai, Kizugawa, Kyoto 619-0292, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             306..1763
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="pCG1 RepA"
                     /protein_id="ADR57710.1"
                     /translation="MTLADSRDIDTPAWKFSADLFDTHPELALRSRGWTSEDRREFLAH
                     LGRENFQGSKTRDFASAWIKDPDTGETQPKLYRVGSKSLARCQYVALTHAQHAAVLVLD
                     IDVPSHQAGGKIEHVNPEVYAILERWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAAAG
                     MSSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYRWHAQHNRVDRLADL
                     MEVARMISGTEKPKKRYEQEFSSGRARIEAARKATAEAKALATLEASLPSAAEASGELI
                     DGVRVLWTAPGRAARDETAFRHALTVGYQLKAAGERLKDTKIIDAYERAYTVAQAVGAD
                     GREPDLPPMRDRQTMARRVRGYVAKGQPVVPARQTETQSSRGRKALATMGRRGGKKAAE
                     RWKDPNSEYARAQREKLAKSSQRQARKAKGNRLTIAGWFMTVEGETGSWPTINEAMSEF
                     SVSRQTVNRALKSAGIELPRGRRKASQ"
     gene            306..1763
                     /gene="repA"
                     /label=repA
     protein_bind    1939..1960
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1975..2005
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2013..2029
                     /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     2037..2053
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2063..2119
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2123..2139)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2361..3173)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3553..4141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatctagca tggtcgtcac agagctggaa gcggcagcga gaattatccg cgatcgtggc
       61 gcggtgcccg caggcatgac aaacatcgta aatgccgcgt ttcgtgtggc cgtggccgcc
      121 caggacgtgt cagcgccgcc accacctgca ccgaatcggc agcagcgtcg cgcgtcgaaa
      181 aagcgcacag gcggcaagaa gcgataagct gcacgaatac ctgaaaaatg ttgaacgccc
      241 cgtgagcggt aactcacagg gcgtcggcta acccccagtc caaacctggg agaaagcgct
      301 caaaaatgac tctagcggat tcacgagaca ttgacacacc ggcctggaaa ttttccgctg
      361 atctgttcga cacccatccc gagctcgcgc tgcgatcacg tggctggacg agcgaagacc
      421 gccgcgaatt cctcgctcac ctgggcagag aaaatttcca gggcagcaag acccgcgact
      481 tcgccagcgc ttggatcaaa gacccggaca cgggagaaac acagccgaag ttataccgag
      541 ttggttcaaa atcgcttgcc cggtgccagt atgttgctct gacgcacgcg cagcacgcag
      601 ccgtgcttgt cctggacatt gatgtgccga gccaccaggc cggcgggaaa atcgagcacg
      661 taaaccccga ggtctacgcg attttggagc gctgggcacg cctggaaaaa gcgccagctt
      721 ggatcggcgt gaatccactg agcgggaaat gccagctcat ctggctcatt gatccggtgt
      781 atgccgcagc aggcatgagc agcccgaata tgcgcctgct ggctgcaacg accgaggaaa
      841 tgacccgcgt tttcggcgct gaccaggctt tttcacatag gctgagccgg tggccactgc
      901 acgtctccga cgatcccacc gcgtaccgct ggcatgccca gcacaatcgc gtggatcgcc
      961 tagctgatct tatggaggtt gctcgcatga tctcaggcac agaaaaacct aaaaaacgct
     1021 atgagcagga gttttctagc ggacgggcac gtatcgaagc ggcaagaaaa gccactgcgg
     1081 aagcaaaagc acttgccacg cttgaagcaa gcctgccgag cgccgctgaa gcgtctggag
     1141 agctgatcga cggcgtccgt gtcctctgga ctgctccagg gcgtgccgcc cgtgatgaga
     1201 cggcttttcg ccacgctttg actgtgggat accagttaaa agcggctggt gagcgcctaa
     1261 aagacaccaa gatcatcgac gcctacgagc gtgcctacac cgtcgctcag gcggtcggag
     1321 cagacggccg tgagcctgat ctgccgccga tgcgtgaccg ccagacgatg gcgcgacgtg
     1381 tgcgcggcta cgtcgctaaa ggccagccag tcgtccctgc tcgtcagaca gagacgcaga
     1441 gcagccgagg gcgaaaagct ctggccacta tgggaagacg tggcggtaaa aaggccgcag
     1501 aacgctggaa agacccaaac agtgagtacg cccgagcaca gcgagaaaaa ctagctaagt
     1561 ccagtcaacg acaagctagg aaagctaaag gaaatcgctt gaccattgca ggttggttta
     1621 tgactgttga gggagagact ggctcgtggc cgacaatcaa tgaagctatg tctgaattta
     1681 gcgtgtcacg tcagaccgtg aatagagcac ttaagtctgc gggcattgaa cttccacgag
     1741 gacgccgtaa agcttcccag taaatgtgcc atctcgtagg cagataacgg ttccccccgt
     1801 aggggtctct ctcttggcct cctttctagg tcgggctgat tgctcttgaa gctctctagg
     1861 ggggctcaca ccataggcag ataacggttc cagatctagg tttcccgact ggaaagcggg
     1921 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     1981 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     2041 aaacagctat gaccatgatt acgaattcga gctcggtacc cggggatcct ctagagtcga
     2101 cctgcaggca tgcaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2161 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2221 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgcg
     2281 atttattcaa caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac
     2341 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     2401 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     2461 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     2521 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     2581 tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag
     2641 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     2701 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     2761 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     2821 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     2881 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     2941 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     3001 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     3061 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     3121 ttggaattta atcgcggctt cgagcaagac gtttcccgtt gaatatggct cataacaccc
     3181 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     3241 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact
     3301 tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca
     3361 aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat caaccgtggc
     3421 tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc agcaacacct
     3481 tcttcacgag gcagacctct cgacggagtt ccactgagcg tcagaccccg tagaaaagat
     3541 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3601 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3661 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3721 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3781 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3841 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3901 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3961 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4021 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4081 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4141 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4201 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4261 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4321 agaagctcgc acattcagca gcgtttttca gcgcgttttc gatcaacgtt tcaatgttgg
     4381 tatcaacacc aggtttaact ttgaacttat cggcactgac ggttactgat tttgaacttt
     4441 tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca
     4501 gcaaaagttc gccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4561 gctggcacg
//
