LOCUS       40924_26719        4806 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pKIL-HIS3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4806)
  AUTHORS   Van Reeth T, Dreze PL, Szpirer J, Szpirer C, Gabant P.
  TITLE     Positive selection vectors to generate fused genes for the 
            expression of his-tagged proteins
  JOURNAL   BioTechniques 25 (5), 898-904 (1998)
  PUBMED    9821593
REFERENCE   2  (bases 1 to 4806)
  AUTHORS   Van Reeth T, Gabant P, Dreze PL, Szpirer J, Szpirer C.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-FEB-1998) Departement de Biologie Moleculaire, 
            Universite Libre de Bruxelles, 67, rue des Chevaux, Rhode St. Genese
            1640, Belgium
REFERENCE   3  (bases 1 to 4806)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4806)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1998"; volume: "25"; issue: "5"; pages: 
            "898-904"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-FEB-1998) Departement de Biologie Moleculaire, Universite Libre 
            de Bruxelles, 67, rue des Chevaux, Rhode St. Genese 1640, Belgium"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..59
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    67..83
                     /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)."
     misc_feature    100..169
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             220..243
                     /label=minicistron
                     /note="synthetic cistron containing a ribosome binding site
                     (Shine-Dalgarno sequence), for enhancing the bacterial 
                     expression of a downstream cistron (Schoner, 1997)"
     CDS             257..271
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             281..298
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             472..771
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     regulatory      876..1002
                     /gene="ccdB"
                     /regulatory_class="terminator"
     terminator      996..1082
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1174..1201
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1221..1312
                     /label=AmpR promoter
     CDS             1313..2170
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2344..2932
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3118..3258)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        3444..3521
                     /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             3522..4601
                     /label=lacI
                     /note="lac repressor"
ORIGIN
        1 ggttctggca aatattctga aatgagctgt tgacaattaa tcatccggct cgtataatgt
       61 gtggaattgt gagcggataa caatttcaca caggaaacag cgccgctgag aaaaagcgaa
      121 gcggcactgc tctttaacaa tttatcagac aatctgtgtg ggcactcgac cggaattatc
      181 gattaacttt attattaaaa attaaagagg tatatattaa tgtatcgatt aaataaggag
      241 gaataaacca tgtcgcgacg acgacgacaa gcgtggttct catcatcatc atcatcatta
      301 atagagggat ccgtcgacct ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
      361 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
      421 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tatacgtacg gcagtttaag
      481 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
      541 attgacacgc cggggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
      601 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
      661 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
      721 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
      781 gaattcgaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
      841 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
      901 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
      961 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1021 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1081 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1141 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1201 ttgcgtttct acaaactctt tttgtttatt tttctaaata cattcaaata tgtatccgct
     1261 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1321 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1381 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1441 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1501 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     1561 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1621 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1681 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1741 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1801 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1861 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1921 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1981 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2041 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2101 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2161 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2221 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2281 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2341 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2401 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2461 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2521 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2581 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2641 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2701 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2761 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2821 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2881 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2941 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3001 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3061 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3121 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     3181 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3241 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3301 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3361 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag
     3421 gcgaagcggc atgcatttac gttgacacca tcgaatggtg caaaaccttt cgcggtatgg
     3481 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     3541 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     3601 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     3661 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     3721 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     3781 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     3841 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     3901 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     3961 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     4021 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     4081 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     4141 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     4201 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     4261 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     4321 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgtcaa
     4381 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     4441 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     4501 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4561 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4621 gtgagttagc gcgaattgat ctggtttgac agcttatcat cgactgcacg gtgcaccaat
     4681 gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact
     4741 gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat
     4801 cataac
//
