LOCUS       sequence-457467-        3808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E.coli genomic integration at arsB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3808)
  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. 2013 Jun 24;12(1):60.
  PUBMED    23799955
REFERENCE   2  (bases 1 to 3808)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb Cell
            Fact."; date: "2013-06-24"; volume: "12(1)"; pages: "60"
COMMENT     Sequence Label: pKIKOarsBCm
FEATURES             Location/Qualifiers
     source          1..3808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      35..423
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(525..1316,1317..1385))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1148..1167
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1386..1490)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    complement(2154..2201)
                     /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)."
     CDS             complement(2218..2877)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2792..2811
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2878..2980)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3086..3119)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 ggaggatatt caaatggacc atggctaatt cccatgtcag ccgttaagtg ttcctgtgtc
       61 actcaaaatt 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 tgatcttcag
      421 atcctctacg ccggacgcat cgtggccgga tcttgcggcc gcaaaaatta aaaatgaagt
      481 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      541 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
      601 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
      661 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
      721 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
      781 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
      841 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
      901 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
      961 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1021 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1081 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1141 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1201 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1261 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1321 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1381 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1441 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     1501 cgaaaagtgc cacctgcatc cagaaacgtc ccaaatcgca gccaatcaca ttggcataaa
     1561 ccatcgcttc tttgataacg ccagatgccg tgctgccatc aatggacaac gcgccaacca
     1621 gtaccgtcgg catattgttc ataatagaag agaggaaggc ggtgaggaat ccggtgccga
     1681 gcgtcgcggc ccacaggccg ttatccgcca gcacgttgag tacgccagaa agatattccg
     1741 ttaatccggc attgcgcagg ccataaacca ccagatacat gccgagcgag aagatgacaa
     1801 tctgccaggg ggcaccgcgc aggactttac ccgtattaat cgcatgaccg cgtttagcga
     1861 cgacaaataa tatcagcgcg cccacagctg caatggcgct caccggactg ccgagcggtt
     1921 ccaggacgaa aaatcccacc agcagaagca gtaaaacaac ccagccagtt ttgaacgtag
     1981 caggatcttt gatcgcttct gcgggagatt tcagcagcgc catatcgtag ttctgcggaa
     2041 tatctttgcg aaaatagaga tgtaacatca ccagcgtggc aacaatcgcg gcgataaaca
     2101 aaaaaaaccc cgcttcggcg gggttttttt taatcgtagg ctggagctgc ttcgaagttc
     2161 ctatactttc tagagaatag gaacttcgga ataggaactt catttaaatg gcgcgcctta
     2221 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     2281 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     2341 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     2401 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     2461 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     2521 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     2581 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     2641 gtctttcatt gccatacgta attccggatg agcattcatc aggcgggcaa gaatgtgaat
     2701 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     2761 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     2821 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     2881 agcttcctta gctcctgaaa atctcgacaa ctcaaaaaat acgcccggta gtgatcttat
     2941 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa
     3001 gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg
     3061 atcttccgtc acaggtaggc gcgccgaagt tcctatactt tctagagaat aggaacttcg
     3121 gaataggaac taaggaggat attcatatag accatgattg catgcggtac cgagctcgga
     3181 tccactagta gaggataaaa catatggaat tcctgcagtg cgcagtcgac gcgcaaaaaa
     3241 ccccgcccct gacagggcgg ggttttttcg ctgtttatcc accggcacca tcaccgaggc
     3301 gtattcgcga aagccgaggc caaagaaatc agcggaaacg atattcacca ggttggagac
     3361 aataagcggc aggctggcgg tatcggcaat gaatccggcc gccatcacga acgccagcgt
     3421 agtgccttta ctgaacccta aagccagcag catggcgatg acaatcggtg tcaaaataag
     3481 cgccgcgcca tcattggcaa acagggcggc aacggcagca ccgagcagga caatccaggt
     3541 aaacagcaag cgaccacgac cattacccca gcgtgagacg tgcagcgccg cccattcaaa
     3601 aaagccggac tcatccagca gcaggctgat gataacgacg gcgataaacg cagccgtcgc
     3661 gttccagacg atattccaca ccaccggaat atcacccgga tggaccacgc ccgtaactaa
     3721 cgccagtact gcgccgagcg ttgcactcca gccgatgcct aaacctttcg gctgccagat
     3781 aaccaatacg atggtcagga cgtttctg
//
