LOCUS       40924_47213        4968 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pXK99CAT, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4968)
  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 4968)
  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 4968)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4968)
  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..4968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..56
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             90..746
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      869..955
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1047..1074
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             1506..2297
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      2400..2988
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3174..3314)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        3500..3577
                     /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             3578..4657
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        4892..4921
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4929..4945
                     /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 caagctgatc
       61 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
      121 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
      181 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
      241 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
      301 cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
      361 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
      421 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
      481 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg
      541 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
      601 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
      661 ttcatcatgc cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
      721 actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa
      781 cgcctggtgc tacgcctgaa taagtgataa taagcggatg aatggcagaa attcggatcc
      841 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
      901 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
      961 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     1021 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
     1081 tctacaaact ctttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     1141 taattccgct agcttcacgc tgccgcaagc actcagggcg caagggctgc taaaggaagc
     1201 ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     1261 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     1321 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     1381 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     1441 cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt
     1501 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     1561 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     1621 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     1681 aactccaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     1741 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     1801 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     1861 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     1921 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     1981 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcggatgc
     2041 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     2101 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     2161 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     2221 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     2281 ttcttgacga gttcttctga gcgggactct ggggttcgcg gaatcatgac caaaatccct
     2341 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2521 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2761 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3001 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3061 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3121 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3181 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     3241 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3301 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3361 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3421 ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga
     3481 agcggcatgc atttacgttg acaccatcga atggtgcaaa acctttcgcg gtatggcatg
     3541 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     3601 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     3661 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     3721 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     3781 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     3841 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     3901 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     3961 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     4021 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     4081 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     4141 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     4201 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     4261 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     4321 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     4381 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgtcaaccac
     4441 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     4501 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     4561 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4621 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     4681 gttagcgcga attgatctgg tttgacagct tatcatcgac tgcacggtgc accaatgctt
     4741 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
     4801 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
     4861 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatccgg ctcgtataat
     4921 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agaccatg
//
