LOCUS       sequence-308428-        2948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zeomycin derivative of lambda-red pKD3 plasmid from Datsenko and 
            Wanner (2000).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2948)
  AUTHORS   Zaworski J, Dagva O, Brandt J, Baum C, Ettwiller L, Fomenkov A, 
            Raleigh EA
  TITLE     Reassembling a cannon in the DNA defense arsenal: Genetics of StySA,
            a BREX phage exclusion system in Salmonella lab strains.
  JOURNAL   PLoS Genet. 2022 Apr 4;18(4):e1009943. doi: 
            10.1371/journal.pgen.1009943. eCollection 2022 Apr.
  PUBMED    35377874
REFERENCE   2  (bases 1 to 2948)
  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: "PLoS 
            Genet."; date: "2022-04-4"; volume: "18(4):e1009943. doi"; pages: " 
            10.1371/journal.pgen.1009943. eCollection 2022 Apr"
COMMENT     Sequence Label: pJBJZ_006
FEATURES             Location/Qualifiers
     source          1..2948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(51..98)
                     /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(358..732)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVVGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQAVPDNTLAWVWVRGLDELYAEWSEVVSTDFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(733..835)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(1089..1122)
                     /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)"
     rep_origin      1171..1559
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(1661..2452,2453..2521))
                     /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     2284..2303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2522..2626)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2716..2802
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2894..2921
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agattgcagc attacacgtc ttgagcgatt gtgtaggctg gagctgcttc gaagttccta
       61 tactttctag agaataggaa cttcggaata ggaacttcca cacaaccaca ccacaccaca
      121 cggggagagc ctgagcaaac tggcctcagg catttgagaa gcacacggtc acactgcttc
      181 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
      241 ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc
      301 acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatca
      361 gtcctgctcc tcggccacga agtgcacgca gttgccggcc gggtcgcgca gggcgaactc
      421 ccgcccccac ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtc
      481 cgtggacacg acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca
      541 ggccagggtg ttgtccggca ccgcctggtc ctggaccgcg ctgatgaaca gggtcacgtc
      601 gtcccggacc acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc
      661 ggtccagaac tcgaccgctc cgacgacgtc gcgcgcggtg agcaccggaa cggcactggt
      721 caacttggcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
      781 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc
      841 tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac caggatttat
      901 ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtgg
      961 tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttggt
     1021 tttggggggt gtttggtgca ggatttattt attctgcgaa gtgatcttcc gtcacaggta
     1081 ggcgcgccga agttcctata ctttctagag aataggaact tcggaatagg aactaaggag
     1141 gatattcata tggaccatgg ctaattccca tgtcagccgt taagtgttcc tgtgtcactc
     1201 aaaattgctt tgagaggctc taagggcttc tcagtgcgtt acatccctgg cttgttgtcc
     1261 acaaccgtta aaccttaaaa gctttaaaag ccttatatat tctttttttt cttataaaac
     1321 ttaaaacctt agaggctatt taagttgctg atttatatta attttattgt tcaaacatga
     1381 gagcttagta cgtgaaacat gagagcttag tacgttagcc atgagagctt agtacgttag
     1441 ccatgagggt ttagttcgtt aaacatgaga gcttagtacg ttaaacatga gagcttagta
     1501 cgtgaaacat gagagcttag tacgtactat caacaggttg aactgctgat cttcagatcc
     1561 tctacgccgg acgcatcgtg gccggatctt gcggccgcaa aaattaaaaa tgaagtttta
     1621 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1681 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1741 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1801 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1861 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1921 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1981 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2041 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2101 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2161 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2221 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2281 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2341 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2401 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2461 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2521 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2581 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2641 aagtgccacc tgcatcgatg gcccccgatg gtagtgtggg gtctccccat gcgagagtag
     2701 ggaactgcca ggcatcaaat aaaatgaaag gctcagtcga aagactgggc ctttcgtttt
     2761 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     2821 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     2881 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtggcc agtgccaagc
     2941 ttgcatgc
//
