LOCUS       40924_26709        4043 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKIKOrbsARCm, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4043)
  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 4043)
  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 4043)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4043)
  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..4043
                     /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..2035)
                     /label=partial rbsR fragment
                     /note="partial rbsR fragment"
     regulatory      2235..2266
                     /label=hairpin loop
                     /note="hairpin loop"
                     /regulatory_class="terminator"
     protein_bind    complement(2289..2336)
                     /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    2289..2330
                     /label=FRT site for Flp/Frt recombination
                     /note="FRT site for Flp/Frt recombination"
     CDS             complement(2356..3012)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3013..3115)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3221..3254)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     regulatory      3367..3406
                     /label=hairpin loop
                     /note="hairpin loop"
                     /regulatory_class="terminator"
     misc_feature    complement(3507..4036)
                     /label=partial rbsA fragment
                     /note="partial rbsA 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 cccagttcat
     1501 ctttcggttg gtggatagtg gttaatggtg gcgtcataaa gcttgccagt tcgatatcgt
     1561 catagccaat caccgcgata tcctgcggaa cctgtaactc tgcctgatat aacgcctggt
     1621 aaacgccaac agccatagcg tcatttccgg taaagacggc ctgaggacgc agcggatgtg
     1681 ataacagttg gcgcatggcg tcaaacccgc cgttaaattc aaaatcacca gtgacttcat
     1741 agccatcagg aatgttgaga cccgcacgtt tcatcgccgc ccgataacct tccaaccgca
     1801 ggcgcgccgg agttttatcc agcggaccgg taatacaggc gatacgggta tgacctttat
     1861 cgatcagata ttgcgttgct aagtctccgc ccagcaacga gttatcctga ataagatcgc
     1921 tgtcgccatc gaacggagcc cagtccatca tcacagtagg cactgtcgga taacgttgca
     1981 tgatttcacg cgaaggttga tgcgtttcgg tgcacagtaa cagccaccag cgcattgact
     2041 tcaatgccga cgatagatgc agcaactgcg ccggttagcg ccaacagaga acctacagac
     2101 aagtcgatgc ccgacgtcag gatcaccagc gtcatcccga ccgccataat ggcgttcacc
     2161 gaggtttgct ggagaatatt gaataagtta ttgatggtga aaaagttcgg gcttaacgtc
     2221 gagacaatcg aaacaaaaaa aaccccgctt cggcggggtt ttttttaatc gtaggctgga
     2281 gctgcttcga agttcctata ctttctagag aataggaact tcggaatagg aacttcattt
     2341 aaatggcgcg ccttacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
     2401 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     2461 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     2521 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
     2581 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     2641 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     2701 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     2761 tatcaccagc tcaccgtctt tcattgccat acgtaattcc ggatgagcat tcatcaggcg
     2821 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     2881 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     2941 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     3001 ttttttctcc attttagctt ccttagctcc tgaaaatctc gacaactcaa aaaatacgcc
     3061 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc
     3121 tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac caggatttat
     3181 ttattctgcg aagtgatctt ccgtcacagg taggcgcgcc gaagttccta tactttctag
     3241 agaataggaa cttcggaata ggaactaagg aggatattca tatagaccat gattgcatgc
     3301 ggtaccgagc tcggatccac tagtagagga taaaacatat ggaattcctg cagtgcgcag
     3361 tcgacgcgca aaaaaccccg cccctgacag ggcggggttt tttcgctgtt tcgatcagca
     3421 ccagcagagc gataagcgat ttctgctcca tcagccacgc tttcgtgaaa taacggcgac
     3481 cagagacagt ctgggttgtc atttttttac tcctgattca cgcgattaag cttgcccaca
     3541 gcggcagcca ttaacacttc ctgggtggcc tgctcacgag taaattcccc gctgagatgc
     3601 ccttcatgca tgacgatgat gcgatcgctc atgcctaata cttctggcat ctccgatgac
     3661 accagaatga tgctcaagcc atcggctttg aactggttaa tcagttgata gatctctttt
     3721 ttcgcgccga catctacgcc acgggtaggt tcatcaagga tcaacacttt ggggcgtgtc
     3781 atcagaccgc gggcaatcgc cactttttgc tgattgccac cggaaagcag accaattgcc
     3841 tgttccatcg acggtgtttt cacattaaac agacgaatga aatcactcac agcctgctgt
     3901 tcatcggcat gcttcaaact gccgccagcg cggctgaagt agcgcagcgc tgtcagcgac
     3961 atgttctctt ttactgacat gcccaacact aaaccgtcac gtttacggtc ttcggagata
     4021 tacacaatgc cgcttggttt ctg
//
