LOCUS       40924_16685        6956 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pEC-XC99E, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6956)
  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 6956)
  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 6956)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6956)
  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..6956
                     /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             716..2179
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="AAO65191.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVGA
                     DGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATSN
                     ARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELAG
                     ELGVSERRIQQVRKALGMEAKRGRPRAEN"
     gene            716..2179
                     /gene="repA"
                     /label=repA
     CDS             2822..3037
                     /codon_start=1
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="AAO65192.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            2822..3037
                     /gene="per"
                     /label=per
     promoter        3464..3566
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             3567..4223
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      4388..4976
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5162..5302)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        5488..5565
                     /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             5566..6645
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        6880..6909
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    6917..6933
                     /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 tagatcccca tcaatcctgc ctatttgcca cgtttaacaa ggtagttaag cgttcattta
      601 cgaagaaaac acgataagct gcacaaatac ctgaaaaagt tgaacgcccc gtgagcggga
      661 actcacaggg cgtcggctaa cccccagtca tcagctggga gaaagcactc aagacatgac
      721 tctagccgat ccgcaggaca cagtcacagc tagcgcgtgg aaattgtccg ccgatctgtt
      781 cgacacccac cccgaagcta tgcgctgcgg ctcacgcggc tggacggcag aagatcgccg
      841 cgaactgctc gctcacctgg gacgcgaaag cttccagggc agcaagacaa gagatttcgc
      901 gagcgcctgg attaaaaacc cggataccgg cgaaacccaa ccaaagctct accgggctgg
      961 ctcaaaagcg ctgacgcggt gccagtacgt tgcgctgacg cacgcgcaac atgccgcggt
     1021 gatcgtgctt gacatcgatg tgcccagcca ccaggccggc gggaagattg agcacgtaaa
     1081 cccgcaggtc tacgcgattt tagagaaatg ggcacgccta gaaaaagcgc cggcttggat
     1141 cggcgtgaat ccgctgagcg ggaaatgcca gctcatctgg ctcattgacc cggtgtatgc
     1201 cgcagcaggt aaaaccagcc caaatatgcg cctgctggct gcaacgacgg aagaaatgac
     1261 tcgtgttttc ggcgctgacc aggctttttc gcataggctg agccggtggc cgctgcacgt
     1321 ctcagacgat ccgacagcct ataaatggca ctgccagcat gatcgtgtgg atcggctggc
     1381 cgacctaatg gagattgctc gaacgatgac cggatcacag aagccgaaaa agtacattga
     1441 gcaggacttt tccagcggac gcgcccgcat tgaagcggca caacgcgcca ccgcagaagc
     1501 caaggcgcta gcgattttgg acgcgagcct gccgagcgcc ctggacgcgt ccggcgacct
     1561 gatcgacggc gtgcgagtgc tctggacaaa tccagagcga gcgcgcgacg agaccgcgtt
     1621 tcgccacgcg ttgaccgtgg gataccagct caaagctgct ggtgagcgcc taaaagatgc
     1681 caagatcatc gacgcgtatg aagtggcgta caacgttgcc caggcggtcg gtgcagacgg
     1741 ccgggagccg gatcttcccg ccatgcgtga tcgcctgacg atggcgcgtc gtgtgcgcgg
     1801 ctacgtggct aaaggccagc cagtcgtccc tgctcgtcgg gtggaaacgc agagcagccg
     1861 agggcggaaa gctctagcga cgatggggcg acggggcgca gctacatcga atgcacgcag
     1921 atgggctgac ccagaaagta agtatgcgca ggagacgcga cagcgattag cggaagcaaa
     1981 caaacgccga gaaatgacag gcgagttgct cgaacttcgc gtcaaaactg cgatcctgga
     2041 tgcccgttct caatcggttg ctgatccctc gactcgtgag cttgcaggcg aactaggtgt
     2101 cagtgaaagg cgcatccaac aagtcagaaa ggcacttgga atggaagcta aacgcggccg
     2161 tccacgggct gaaaactaat aaacgaaaca ccgtcagcag aaaacggttc ccccctttag
     2221 gggtcccgtc cttgctctgg ctctcacttg ccctcaccct ccgctatcca cgggctgaaa
     2281 actaataaac gaaacaccgt cagcagaaaa cggttccccc cctttagggt gtctcgctcc
     2341 tagctctgat ccctccccgg ttcctccccg gcctgatttt taaggggggc tcacgctgtc
     2401 ggcagagaac ggttccccgc cttctgctct ggctcttcct cgactccctc cccctcaaaa
     2461 atctcctcga gatcctggag acctttttgg agctagcgcg ttgctgcttc gcaccaactt
     2521 gctcatgatg attttcattt ttgcttgtgt gcttttttgg gttgaaccct ccaaagaggg
     2581 gaaaccaggg gcacacctca tgcactaaag tgccgcttcg ctggtcaggg tgaaatcacc
     2641 tggaaaaaaa gtgcggtaac cgctgcgctt ggcgtttttt ctgggcaaga agtctcgcag
     2701 gttttcgcag gagtgccgga agaaattatc agaattgggg ctagaatttt taacgaacgt
     2761 tcgttataat ggtgtcatga ccttcacgac gaagtaccaa aactggcctg aagcatcagc
     2821 ggtggatctc tccgatgtcg cgctggagtc cgacgcactc gatgccgccg tcgatttaaa
     2881 aacggtgatc ggatttttcc gcgccctcga tacgacagac gcgccagcat cacgcgactg
     2941 ggcaagtgcc gcgagcgacc tagaaacgct tgtggccgac cttgaagagc tggccgacga
     3001 gctgcgtgct cggcagcgcc aggaggacgc gcagtagtgg aggatcgcat cagctgcgcc
     3061 tactgcggtg gcctgatccc accccggcct gacccacgag gacggcgcgc aaaatactgc
     3121 tcagacgcgt gtcgtgccgc agccagccgc gagcgcgcca acaagcgcca cgcccaggag
     3181 gtcgaagccg cacgtcatct agcgctgatg tccggcggtg cttttgccgt tacgcaccac
     3241 cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg gagcacctca
     3301 aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg
     3361 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
     3421 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag
     3481 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     3541 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     3601 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     3661 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     3721 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     3781 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     3841 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     3901 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     3961 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     4021 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     4081 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     4141 atcatgccgt ctgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     4201 gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc
     4261 ctggtgctac gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgcatgacca
     4321 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4381 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4441 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4501 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4561 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4621 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4681 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4741 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4801 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4861 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4921 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4981 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5041 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5101 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5161 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     5221 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     5281 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     5341 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5401 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
     5461 gaaggcgaag cggcatgcat ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt
     5521 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     5581 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     5641 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     5701 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     5761 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     5821 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     5881 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     5941 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     6001 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     6061 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     6121 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     6181 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     6241 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     6301 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     6361 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     6421 tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     6481 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     6541 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6601 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6661 taatgtgagt tagcgcgaat tgatctggtt tgacagctta tcatcgactg cacggtgcac
     6721 caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat
     6781 cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg
     6841 acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa tcatccggct
     6901 cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag accatg
//
