LOCUS       40924_16690        7018 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pEC-XK99E, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  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 7018)
  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 7018)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7018)
  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..7018
                     /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(705..920)
                     /codon_start=1
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="AAO65195.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            complement(705..920)
                     /gene="per"
                     /label=per
     CDS             complement(1563..3026)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="AAO65196.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVGA
                     DGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATSN
                     ARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELAG
                     ELGVSERRIQQVRKALGMEAKRGRPRAEN"
     gene            complement(1563..3026)
                     /gene="repA"
                     /label=repA
     CDS             3556..4347
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      4450..5038
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5224..5364)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        5550..5627
                     /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             5628..6707
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        6942..6971
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    6979..6995
                     /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 tctttttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattccgc
      541 tagatgacgt gcggcttcga cctcctgggc gtggcgcttg ttggcgcgct cgcggctggc
      601 tgcggcacga cacgcgtctg agcagtattt tgcgcgccgt cctcgtgggt caggccgggg
      661 tgggatcagg ccaccgcagt aggcgcagct gatgcgatcc tccactactg cgcgtcctcc
      721 tggcgctgcc gagcacgcag ctcgtcggcc agctcttcaa ggtcggccac aagcgtttct
      781 aggtcgctcg cggcacttgc ccagtcgcgt gatgctggcg cgtctgtcgt atcgagggcg
      841 cggaaaaatc cgatcaccgt ttttaaatcg acggcggcat cgagtgcgtc ggactccagc
      901 gcgacatcgg agagatccac cgctgatgct tcaggccagt tttggtactt cgtcgtgaag
      961 gtcatgacac cattataacg aacgttcgtt aaaaattcta gccccaattc tgataatttc
     1021 ttccggcact cctgcgaaaa cctgcgagac ttcttgccca gaaaaaacgc caagcgcagc
     1081 ggttaccgca ctttttttcc aggtgatttc accctgacca gcgaagcggc actttagtgc
     1141 atgaggtgtg cccctggttt cccctctttg gagggttcaa cccaaaaaag cacacaagca
     1201 aaaatgaaaa tcatcatgag caagttggtg cgaagcagca acgcgctagc tccaaaaagg
     1261 tctccaggat ctcgaggaga tttttgaggg ggagggagtc gaggaagagc cagagcagaa
     1321 ggcggggaac cgttctctgc cgacagcgtg agcccccctt aaaaatcagg ccggggagga
     1381 accggggagg gatcagagct aggagcgaga caccctaaag ggggggaacc gttttctgct
     1441 gacggtgttt cgtttattag ttttcagccc gtggatagcg gagggtgagg gcaagtgaga
     1501 gccagagcaa ggacgggacc cctaaagggg ggaaccgttt tctgctgacg gtgtttcgtt
     1561 tattagtttt cagcccgtgg acggccgcgt ttagcttcca ttccaagtgc ctttctgact
     1621 tgttggatgc gcctttcact gacacctagt tcgcctgcaa gctcacgagt cgagggatca
     1681 gcaaccgatt gagaacgggc atccaggatc gcagttttga cgcgaagttc gagcaactcg
     1741 cctgtcattt ctcggcgttt gtttgcttcc gctaatcgct gtcgcgtctc ctgcgcatac
     1801 ttactttctg ggtcagccca tctgcgtgca ttcgatgtag ctgcgccccg tcgccccatc
     1861 gtcgctagag ctttccgccc tcggctgctc tgcgtttcca cccgacgagc agggacgact
     1921 ggctggcctt tagccacgta gccgcgcaca cgacgcgcca tcgtcaggcg atcacgcatg
     1981 gcgggaagat ccggctcccg gccgtctgca ccgaccgcct gggcaacgtt gtacgccact
     2041 tcatacgcgt cgatgatctt ggcatctttt aggcgctcac cagcagcttt gagctggtat
     2101 cccacggtca acgcgtggcg aaacgcggtc tcgtcgcgcg ctcgctctgg atttgtccag
     2161 agcactcgca cgccgtcgat caggtcgccg gacgcgtcca gggcgctcgg caggctcgcg
     2221 tccaaaatcg ctagcgcctt ggcttctgcg gtggcgcgtt gtgccgcttc aatgcgggcg
     2281 cgtccgctgg aaaagtcctg ctcaatgtac tttttcggct tctgtgatcc ggtcatcgtt
     2341 cgagcaatct ccattaggtc ggccagccga tccacacgat catgctggca gtgccattta
     2401 taggctgtcg gatcgtctga gacgtgcagc ggccaccggc tcagcctatg cgaaaaagcc
     2461 tggtcagcgc cgaaaacacg agtcatttct tccgtcgttg cagccagcag gcgcatattt
     2521 gggctggttt tacctgctgc ggcatacacc gggtcaatga gccagatgag ctggcatttc
     2581 ccgctcagcg gattcacgcc gatccaagcc ggcgcttttt ctaggcgtgc ccatttctct
     2641 aaaatcgcgt agacctgcgg gtttacgtgc tcaatcttcc cgccggcctg gtggctgggc
     2701 acatcgatgt caagcacgat caccgcggca tgttgcgcgt gcgtcagcgc aacgtactgg
     2761 caccgcgtca gcgcttttga gccagcccgg tagagctttg gttgggtttc gccggtatcc
     2821 gggtttttaa tccaggcgct cgcgaaatct cttgtcttgc tgccctggaa gctttcgcgt
     2881 cccaggtgag cgagcagttc gcggcgatct tctgccgtcc agccgcgtga gccgcagcgc
     2941 atagcttcgg ggtgggtgtc gaacagatcg gcggacaatt tccacgcgct agctgtgact
     3001 gtgtcctgcg gatcggctag agtcatgtct tgagtgcttt ctcccagctg atgactgggg
     3061 gttagccgac gccctgtgag ttcccgctca cggggcgttc aactttttca ggtatttgtg
     3121 cagcttatcg tgttttcttc gtaaatgaac gcttaactac cttgttaaac gtggcaaata
     3181 ggcaggattg atggggatct agcttcacgc tgccgcaagc actcagggcg caagggctgc
     3241 taaaggaagc ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat
     3301 gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct
     3361 tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg
     3421 gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg
     3481 gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga
     3541 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3601 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3661 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3721 gccctgaatg aactccaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3781 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3841 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3901 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3961 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4021 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4081 gcgcggatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4141 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4201 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4261 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4321 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg gaatcatgac
     4381 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4441 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4501 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4561 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4621 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4681 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4741 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4801 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4861 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4921 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4981 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5041 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5101 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5161 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5221 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     5281 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     5341 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     5401 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5461 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc
     5521 gcgaaggcga agcggcatgc atttacgttg acaccatcga atggtgcaaa acctttcgcg
     5581 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     5641 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     5701 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     5761 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     5821 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     5881 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     5941 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     6001 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     6061 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     6121 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     6181 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     6241 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     6301 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     6361 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     6421 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     6481 cgtcaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     6541 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     6601 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6661 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6721 attaatgtga gttagcgcga attgatctgg tttgacagct tatcatcgac tgcacggtgc
     6781 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     6841 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     6901 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatccgg
     6961 ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac agaccatg
//
