LOCUS       Exported                7017 bp DNA     circular SYN 02-SEP-2024
DEFINITION  Shuttle expression vector pEC-XK99E, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7017)
  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 7017)
  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 7017)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7017)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7017)
  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"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..57
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      260..346
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      438..465
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(704..919)
                     /codon_start=1
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="AAO65195.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            complement(704..919)
                     /gene="per"
                     /label=per
     CDS             complement(1562..3025)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="AAO65196.1"
                     /translation="MTLADPQDTVTASAWKFSADLFDTHPELALRSRGWTAEDRRELLA
                     HLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIVL
                     DIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAAA
                     GKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLAD
                     LMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGDL
                     IDGVRVLWTNPERAARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVGA
                     DGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATSN
                     ARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELAG
                     ELGVSERRIQQVRKALGMEAKRGRPRAEN"
     gene            complement(1562..3025)
                     /gene="repA"
                     /label=repA
     CDS             3555..4346
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      4449..5037
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5223..5363)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        5549..5626
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5627..6706
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        6941..6970
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    6978..6994
                     /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)."
ORIGIN
        1 gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc
       61 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
      121 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
      181 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
      241 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      301 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
      361 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
      421 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
      481 tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattccgct
      541 agatgacgtg cggcttcgac ctcctgggcg tggcgcttgt tggcgcgctc gcggctggct
      601 gcggcacgac acgcgtctga gcagtatttt gcgcgccgtc ctcgtgggtc aggccggggt
      661 gggatcaggc caccgcagta ggcgcagctg atgcgatcct ccactactgc gcgtcctcct
      721 ggcgctgccg agcacgcagc tcgtcggcca gctcttcaag gtcggccaca agcgtttcta
      781 ggtcgctcgc ggcacttgcc cagtcgcgtg atgctggcgc gtctgtcgta tcgagggcgc
      841 ggaaaaatcc gatcaccgtt tttaaatcga cggcggcatc gagtgcgtcg gactccagcg
      901 cgacatcgga gagatccacc gctgatgctt caggccagtt ttggtacttc gtcgtgaagg
      961 tcatgacacc attataacga acgttcgtta aaaattctag ccccaattct gataatttct
     1021 tccggcactc ctgcgaaaac ctgcgagact tcttgcccag aaaaaacgcc aagcgcagcg
     1081 gttaccgcac tttttttcca ggtgatttca ccctgaccag cgaagcggca ctttagtgca
     1141 tgaggtgtgc ccctggtttc ccctctttgg agggttcaac ccaaaaaagc acacaagcaa
     1201 aaatgaaaat catcatgagc aagttggtgc gaagcagcaa cgcgctagct ccaaaaagct
     1261 ctccaggatc tcgaggagat ttttgagggg gagggagtcg aggaagagcc agagcagaag
     1321 gcgggggaac cgttctctgc cgacagcgtg agccccctta aaaatcaggc cggggaggaa
     1381 ccggggaggg atcagagcta ggagcgagac accctaaagg gggggaaccg ttttctgctg
     1441 acggtgtttc gtttattagt tttcagcccg tggatagcgg agggtgaggg caagtgagag
     1501 ccagagcaag gacgggaccc ctaaaggggg gaaccgtttt ctgctgacgg tgtttcgttt
     1561 attagttttc agcccgtgga cggccgcgtt tagcttccat tccaagtgcc tttctgactt
     1621 gttggatgcg cctttcactg acacctagtt cgcctgcaag ctcacgagtc gagggatcag
     1681 caaccgattg agaacgggca tccaggatcg cagttttgac gcgaagttcg agcaactcgc
     1741 ctgtcatttc tcggcgtttg tttgcttccg ctaatcgctg tcgcgtctcc tgcgcatact
     1801 tactttctgg gtcagcccat ctgcgtgcat tcgatgtagc tgcgccccgt cgccccatcg
     1861 tcgctagagc tttccgccct cggctgctct gcgtttccac ccgacgagca gggacgactg
     1921 gctggccttt agccacgtag ccgcgcacac gacgcgccat cgtcaggcga tcacgcatgg
     1981 cgggaagatc cggctcccgg ccgtctgcac cgaccgcctg ggcaacgttg tacgccactt
     2041 catacgcgtc gatgatcttg gcatctttta ggcgctcacc agcagctttg agctggtatc
     2101 ccacggtcaa cgcgtggcga aacgcggtct cgtcgcgcgc tgcgcgctct ggatttgtcc
     2161 agagcactcg cacgccgtcg atcaggtcgc cggacgcgtc cagggcgctc ggcaggctcg
     2221 cgtccaaaat cgctagcgcc ttggcttctg cggtggcgcg ttgtgccgct tcaatgcggg
     2281 cgcgtccgct ggaaaagtcc tgctcaatgt actttttcgg cttctgtgat ccggtcatcg
     2341 ttcgagcaat ctccattagg tcggccagcc gatccacacg atcatgctgg cagtgccatt
     2401 tataggctgt cggatcgtct gagacgtgca gcggccaccg gctcagccta tgcgaaaaag
     2461 cctggtcagc gccgaaaaca cgagtcattt cttccgtcgt tgcagccagc aggcgcatat
     2521 ttgggctggt tttacctgct gcggcataca ccgggtcaat gagccagatg agctggcatt
     2581 tcccgctcag cggattcacg ccgatccaag ccggcgcttt ttctaggcgt gcccatttct
     2641 ctaaaatcgc gtagacctgc gggtttacgt gctcaatctt cccgccggcc tggtggctgg
     2701 gcacatcgat gtcaagcacg atcaccgcgg catgttgcgc gtgcgtcagc gcaacgtact
     2761 ggcaccgcgt cagcgctttt gagccagccc ggtagagctt tggttgggtt tcgccggtat
     2821 ccgggttttt aatccaggcg ctcgcgaaat ctcttgtctt gctgccctgg aagctttcgc
     2881 gtcccaggtg agcgagcagt tcgcggcgat cttctgccgt ccagccgcgt gagcgcagcg
     2941 ctagttcggg gtgggtgtcg aacagatcgg cggaaaattt ccacgcgcta gctgtgactg
     3001 tgtcctgcgg atcggctaga gtcatgtctt gagtgctttc tcccagctga tgactggggg
     3061 ttagccgacg ccctgtgagt tcccgctcac ggggcgttca actttttcag gtatttgtgc
     3121 agcttatcgt gttttcttcg taaatgaacg cttaactacc ttgttaaacg tggcaaatag
     3181 gcaggattga tggggatcta gcttcacgct gccgcaagca ctcagggcgc aagggctgct
     3241 aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg
     3301 tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt
     3361 gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg
     3421 aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg
     3481 ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat
     3541 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3601 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3661 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3721 ccctgaatga actccaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3781 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3841 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3901 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3961 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4021 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4081 cgcggatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4141 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4201 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4261 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4321 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcgg aatcatgacc
     4381 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4441 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4501 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4561 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4621 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4681 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4741 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4801 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4861 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4921 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4981 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5041 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5101 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5161 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5221 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     5281 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     5341 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     5401 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5461 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagcagatc aattcgcgcg
     5521 cgaaggcgaa gcggcatgca tttacgttga caccatcgaa tggtgcaaaa cctttcgcgg
     5581 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     5641 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     5701 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     5761 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     5821 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     5881 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     5941 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     6001 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     6061 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     6121 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     6181 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     6241 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     6301 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     6361 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     6421 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     6481 gtcaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     6541 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     6601 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6661 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6721 ttaatgtgag ttagcgcgaa ttgatctggt ttgacagctt atcatcgact gcacggtgca
     6781 ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa
     6841 tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt tttttgcgcc
     6901 gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta atcatccggc
     6961 tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca gaccatg
//
