LOCUS       40924_26699        3696 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKIKOlacZCm, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3696)
  AUTHORS   Sabri S, Steen JA, Bongers M, Nielsen LK, Vickers CE.
  TITLE     Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA
            sequences, including whole metabolic pathways, onto the Escherichia 
            coli chromosome at well-characterised loci
  JOURNAL   Microb. Cell Fact. 12 (1), 60 (2013)
  PUBMED    23799955
REFERENCE   2  (bases 1 to 3696)
  AUTHORS   Steen JA, Sabri S, Bongers M, Nielsen LK, Vickers CE.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2013) Australian Institute for Bioengineering and 
            Nanotechnology, University of Queensland, Crn College and Cooper 
            Rds, St Lucia, QLD 4072, Australia
REFERENCE   3  (bases 1 to 3696)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3696)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2013"; volume: "12"; issue: "1"; pages: "60"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-JAN-2013) Australian Institute for Bioengineering and 
            Nanotechnology, University of Queensland, Crn College and Cooper 
            Rds, St Lucia, QLD 4072, Australia"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(35..418)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(491..1348)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1349..1453)
                     /label=AmpR promoter
     misc_feature    complement(1491..2000)
                     /label=partial lacZ fragment
                     /note="partial lacZ fragment"
     regulatory      2008..2039
                     /label=hairpin loop
                     /note="hairpin loop"
                     /regulatory_class="terminator"
     protein_bind    complement(2062..2109)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    2062..2103
                     /label=FRT site for Flp/Frt recombination
                     /note="FRT site for Flp/Frt recombination"
     CDS             complement(2129..2785)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(2786..2888)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2994..3027)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     regulatory      3140..3179
                     /label=hairpin loop
                     /note="hairpin loop"
                     /regulatory_class="terminator"
     misc_feature    complement(3185..3688)
                     /label=partial lacZ fragment
                     /note="partial lacZ fragment"
ORIGIN
        1 ggaggatatt catatggacc atggctaatt cccatgtcag ccgttaagtg ttcctgtgtc
       61 actgaaaatt gctttgagag gctctaaggg cttctcagtg cgttacatcc ctggcttgtt
      121 gtccacaacc gttaaacctt aaaagcttta aaagccttat atattctttt ttttcttata
      181 aaacttaaaa ccttagaggc tatttaagtt gctgatttat attaatttta ttgttcaaac
      241 atgagagctt agtacgtgaa acatgagagc ttagtacgtt agccatgaga gcttagtacg
      301 ttagccatga gggtttagtt cgttaaacat gagagcttag tacgttaaac atgagagctt
      361 agtacgtgaa acatgagagc ttagtacgta ctatcaacag gttgaactgc ggatcttgcg
      421 gccgcaaaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
      481 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
      541 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
      601 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
      661 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
      721 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
      781 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
      841 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
      901 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
      961 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1021 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     1081 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     1141 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     1201 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     1261 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     1321 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     1381 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     1441 taggggttcc gcgcacattt ccccgaaaag tgccacctgc atccagaaac aagactgtta
     1501 cccatcgcgt gggcgtattc gcaaaggatc agcgggcgca tctctccagg tagcgaaagc
     1561 cattttttga tggaccattt cggcacagcc gggaagggct ggtcttcatc cacgcgcgcg
     1621 tacatcgggc aaataatatc ggtggccgtg gtgtcggctc cgccgccttc atactgcacc
     1681 gggcgggaag gatcgacaga tttgatccag cgatacagcg cgtcgtgatt agcgccgtgg
     1741 cctgattcat tccccagcga ccagatgatc acactcgggt gattacgatc gcgctgcacc
     1801 attcgcgtta cgcgttcgct catcgccggt agccagcgcg gatcatcggt cagacgattc
     1861 attggcacca tgccgtgggt ttcaatattg gcttcatcca ccacatacag gccgtagcgg
     1921 tcgcacagcg tgtaccacag cggatggttc ggataatgcg aacagcgcac ggcgttaaag
     1981 ttgttctgct tcatcagcag gataaacaaa aaaaaccccg cttcggcggg gtttttttta
     2041 atcgtaggct ggagctgctt cgaagttcct atactttcta gagaatagga acttcggaat
     2101 aggaacttca tttaaatggc gcgccttacg ccccgccctg ccactcatcg cagtactgtt
     2161 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     2221 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     2281 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     2341 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     2401 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     2461 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     2521 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgtaat tccggatgag
     2581 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     2641 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     2701 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     2761 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgacaact
     2821 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     2881 gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga
     2941 caccaggatt tatttattct gcgaagtgat cttccgtcac aggtaggcgc gccgaagttc
     3001 ctatactttc tagagaatag gaacttcgga ataggaacta aggaggatat tcatatagac
     3061 catgattgca tgcggtaccg agctcggatc cactagtaga ggataaaaca tatggaattc
     3121 ctgcagtgcg cagtcgacgc gcaaaaaacc ccgcccctga cagggcgggg ttttttcgct
     3181 gtttatcctg caccatcgtc tgctcatcca tgacctgacc atgcagagga tgatgctcgt
     3241 gacggttaac gcctcgaatc agcaacggct tgccgttcag cagcagcaga ccattttcaa
     3301 tccgcacctc gcggaaaccg acatcgcagg cttctgcttc aatcagcgtg ccgtcggcgg
     3361 tgtgcagttc aaccaccgca cgatagagat tcgggatttc ggcgctccac agcgccgggt
     3421 tttcgacgtt cagacgtagc gtgacgcgat cggcataacc accacgctca tcgataattt
     3481 caccgccgaa aggcgcggtg ccgctggcga cctgcgtttc accctgccac aaagaaaccg
     3541 tcacccgcag ctcatcgcgc agctcgccgt acatctgaac ttctgcctcc agtaccgcgc
     3601 ggctgaaatc atcattaaag tgagtggcaa catggaaatc gctgatttgt gtgctcggtt
     3661 tgtgcagcaa cgagacgtca cggaaaatgg tttctg
//
