LOCUS       40924_47123        4297 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pXC99E, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4297)
  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 4297)
  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 4297)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4297)
  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..4297
                     /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"
     promoter        805..907
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             908..1564
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      1729..2317
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2503..2643)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        2829..2906
                     /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             2907..3986
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        4221..4250
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4258..4274
                     /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 tagcgctgat gtccggcggt gcttttgccg ttacgcacca ccccgtcagt agctgaacag
      601 gagggacagc tgatagaaac agaagccact ggagcacctc aaaaacacca tcatacacta
      661 aatcagtaag ttggcagcat cacccgacgc actttgcgcc gaataaatac ctgtgacgga
      721 agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga agccctgggc
      781 caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact ttcaccataa
      841 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
      901 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
      961 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1021 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1081 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     1141 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     1201 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     1261 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     1321 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     1381 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     1441 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tctgtgatgg
     1501 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1561 ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggtgcta cgcctgaata
     1621 agtgataata agcggatgaa tggcagaaat tcgcatgacc aaaatccctt aacgtgagtt
     1681 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1741 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1801 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1861 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1921 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1981 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2041 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2101 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2161 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2221 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2281 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2341 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2401 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2461 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2521 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     2581 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     2641 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2701 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2761 tcatcaccga aacgcgcgag gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca
     2821 tttacgttga caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg
     2881 aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt
     2941 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
     3001 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
     3061 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
     3121 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
     3181 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
     3241 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     3301 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     3361 accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
     3421 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     3481 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
     3541 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
     3601 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
     3661 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
     3721 gatacgacga taccgaagac agctcatgtt atatcccgcc gtcaaccacc atcaaacagg
     3781 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     3841 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc
     3901 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3961 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagcgcgaa
     4021 ttgatctggt ttgacagctt atcatcgact gcacggtgca ccaatgcttc tggcgtcagg
     4081 cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg
     4141 ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc
     4201 aaatattctg aaatgagctg ttgacaatta atcatccggc tcgtataatg tgtggaattg
     4261 tgagcggata acaatttcac acaggaaaca gaccatg
//
