LOCUS       40924_16655        5869 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pEC-K19MECA2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5869)
  AUTHORS   Kirchner O, Tauch A.
  TITLE     Tools for genetic engineering in the amino acid-producing bacterium 
            Corynebacterium glutamicum
  JOURNAL   J. Biotechnol. 104 (1-3), 287-299 (2003)
  PUBMED    12948646
REFERENCE   2  (bases 1 to 5869)
  AUTHORS   Kirchner O, Tauch A.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JAN-2003) Department of Genetics, University of 
            Bielefeld, Universitaetsstrasse 25, Bielefeld D-33615, Germany
REFERENCE   3  (bases 1 to 5869)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5869)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Biotechnol. 104 (1-3), 287-299 (2003)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JAN-2003) Department of Genetics, University of Bielefeld, 
            Universitaetsstrasse 25, Bielefeld D-33615, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(206..223)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(232..248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             574..2037
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="AAO65228.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVGA
                     DGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATSN
                     ARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELAG
                     ELGVSERRIQQVRKALGMEAKRGRPRAEN"
     gene            574..2037
                     /gene="repA"
                     /label=repA
     CDS             2680..2895
                     /codon_start=1
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="AAO65229.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            2680..2895
                     /gene="per"
                     /label=per
     CDS             3414..4205
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     oriT            complement(4546..4655)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      4867..5455
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5743..5764
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5779..5809
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5817..5833
                     /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     5841..5857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagctttaat acgactcact atagggagat gttaaccgcg gtccctaggc cggccgctag
       61 ccatttaaat gcatgctgca gcccgggcga cgtcgactgt ttaaacgagg ccttaattaa
      121 gggcccaatt ggccacgtac gtctagatct ggcgcgcctc gaggatccat ggtaccggtg
      181 cggccgcact agtgatcgat atctcccttt agtgagggtt aatgtgaatt cactggccgt
      241 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      301 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      361 acagttgcgc agcctgaatg gcgaatggcg cgataagcta gatccccatc aatcctgcct
      421 atttgccacg tttaacaagg tagttaagcg ttcatttacg aagaaaacac gataagctgc
      481 acaaatacct gaaaaagttg aacgccccgt gagcgggaac tcacagggcg tcggctaacc
      541 cccagtcatc agctgggaga aagcactcaa gacatgactc tagccgatcc gcaggacaca
      601 gtcacagcta gcgcgtggaa attgtccgcc gatctgttcg acacccaccc cgaagctatg
      661 cgctgcggct cacgcggctg gacggcagaa gatcgccgcg aactgctcgc tcacctggga
      721 cgcgaaagct tccagggcag caagacaaga gatttcgcga gcgcctggat taaaaacccg
      781 gataccggcg aaacccaacc aaagctctac cgggctggct caaaagcgct gacgcggtgc
      841 cagtacgttg cgctgacgca cgcgcaacat gccgcggtga tcgtgcttga catcgatgtg
      901 cccagccacc aggccggcgg gaagattgag cacgtaaacc cgcaggtcta cgcgatttta
      961 gagaaatggg cacgcctaga aaaagcgccg gcttggatcg gcgtgaatcc gctgagcggg
     1021 aaatgccagc tcatctggct cattgacccg gtgtatgccg cagcaggtaa aaccagccca
     1081 aatatgcgcc tgctggctgc aacgacggaa gaaatgactc gtgttttcgg cgctgaccag
     1141 gctttttcgc ataggctgag ccggtggccg ctgcacgtct cagacgatcc gacagcctat
     1201 aaatggcact gccagcatga tcgtgtggat cggctggccg acctaatgga gattgctcga
     1261 acgatgaccg gatcacagaa gccgaaaaag tacattgagc aggacttttc cagcggacgc
     1321 gcccgcattg aagcggcaca acgcgccacc gcagaagcca aggcgctagc gattttggac
     1381 gcgagcctgc cgagcgccct ggacgcgtcc ggcgacctga tcgacggcgt gcgagtgctc
     1441 tggacaaatc cagagcgagc gcgcgacgag accgcgtttc gccacgcgtt gaccgtggga
     1501 taccagctca aagctgctgg tgagcgccta aaagatgcca agatcatcga cgcgtatgaa
     1561 gtggcgtaca acgttgccca ggcggtcggt gcagacggcc gggagccgga tcttcccgcc
     1621 atgcgtgatc gcctgacgat ggcgcgtcgt gtgcgcggct acgtggctaa aggccagcca
     1681 gtcgtccctg ctcgtcgggt ggaaacgcag agcagccgag ggcggaaagc tctagcgacg
     1741 atggggcgac ggggcgcagc tacatcgaat gcacgcagat gggctgaccc agaaagtaag
     1801 tatgcgcagg agacgcgaca gcgattagcg gaagcaaaca aacgccgaga aatgacaggc
     1861 gagttgctcg aacttcgcgt caaaactgcg atcctggatg cccgttctca atcggttgct
     1921 gatccctcga ctcgtgagct tgcaggcgaa ctaggtgtca gtgaaaggcg catccaacaa
     1981 gtcagaaagg cacttggaat ggaagctaaa cgcggccgtc cacgggctga aaactaataa
     2041 acgaaacacc gtcagcagaa aacggttccc ccctttaggg gtcccgtcct tgctctggct
     2101 ctcacttgcc ctcaccctcc gctatccacg ggctgaaaac taataaacga aacaccgtca
     2161 gcagaaaacg gttccccccc tttagggtgt ctcgctccta gctctgatcc ctccccggtt
     2221 cctccccggc ctgattttta aggggggctc acgctgtcgg cagagaacgg ttccccgcct
     2281 tctgctctgg ctcttcctcg actccctccc cctcaaaaat ctcctcgaga tcctggagac
     2341 ctttttggag ctagcgcgtt gctgcttcgc accaacttgc tcatgatgat tttcattttt
     2401 gcttgtgtgc ttttttgggt tgaaccctcc aaagagggga aaccaggggc acacctcatg
     2461 cactaaagtg ccgcttcgct ggtcagggtg aaatcacctg gaaaaaaagt gcggtaaccg
     2521 ctgcgcttgg cgttttttct gggcaagaag tctcgcaggt tttcgcagga gtgccggaag
     2581 aaattatcag aattggggct agaattttta acgaacgttc gttataatgg tgtcatgacc
     2641 ttcacgacga agtaccaaaa ctggcctgaa gcatcagcgg tggatctctc cgatgtcgcg
     2701 ctggagtccg acgcactcga tgccgccgtc gatttaaaaa cggtgatcgg atttttccgc
     2761 gccctcgata cgacagacgc gccagcatca cgcgactggg caagtgccgc gagcgaccta
     2821 gaaacgcttg tggccgacct tgaagagctg gccgacgagc tgcgtgctcg gcagcgccag
     2881 gaggacgcgc agtagtggag gatcgcatca gctgcgccta ctgcggtggc ctgatcccac
     2941 cccggcctga cccacgagga cggcgcgcaa aatactgctc agacgcgtgt cgtgccgcag
     3001 ccagccgcga gcgcgccaac aagcgccacg cccaggaggt cgaagccgca cgtcatctag
     3061 cttcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg aacacgtaga
     3121 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     3181 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     3241 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     3301 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     3361 gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3421 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3481 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3541 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctccaagacg
     3601 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3661 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3721 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3781 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3841 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3901 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcggatgccc gacggcgagg
     3961 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4021 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4081 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4141 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4201 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4261 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4321 ggacgccctc gcggacgtgc tcatagtcca cgacgcccgt gattttgtag ccctggccga
     4381 cggccagcag gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc
     4441 ttcgttcgtc tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg
     4501 tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca
     4561 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
     4621 cgctcgcggg tgggcctact tcacctatcc tgccccgctg acgccgttgg atacaccaag
     4681 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
     4741 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaag ccatgaccaa
     4801 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4861 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4921 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4981 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5041 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5101 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5161 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5221 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     5281 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5341 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5401 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5461 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5521 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5581 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5641 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5701 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5761 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5821 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcc
//
