LOCUS       sequence-330061-        2946 bp    DNA     circular SYN 14-APR-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 2946)
  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 2946)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..2946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      22..49
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    complement(127..174)
                     /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(433..807)
                     /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(808..910)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(1163..1196)
                     /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      1245..1633
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(1735..2526,2527..2595))
                     /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     2358..2377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2596..2700)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2790..2876
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtggccag
       61 tgccaagctt gcatgcagat tgcagcatta cacgtcttga gcgattgtgt aggctggagc
      121 tgcttcgaag ttcctatact ttctagagaa taggaacttc ggaataggaa cttccacaca
      181 accacaccac accacacggg agagcctgag caaactggcc tcaggcattt gagaagcaca
      241 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
      301 tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt
      361 catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact
      421 gccttaaaaa aatcagtcct gctcctcggc cacgaagtgc acgcagttgc cggccgggtc
      481 gcgcagggcg aactcccgcc cccacggctg ctcgccgatc tcggtcatgg ccggcccgga
      541 ggcgtcccgg aagtccgtgg acacgacctc cgaccactcg gcgtacagct cgtccaggcc
      601 gcgcacccac acccaggcca gggtgttgtc cggcaccgcc tggtcctgga ccgcgctgat
      661 gaacagggtc acgtcgtccc ggaccacacc ggcgaagtcg tcctccacga agtcccggga
      721 gaacccgagc cggtcggtcc agaactcgac cgctccgacg acgtcgcgcg cggtgagcac
      781 cggaacggca ctggtcaact tggccatttt agcttcctta gctcctgaaa atctcgataa
      841 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
      901 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
      961 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
     1021 aagtgcgtcg ggtggtgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg
     1081 ctccagtggc ttggttttgg gggtgtttgg tgcaggattt atttattctg cgaagtgatc
     1141 ttccgtcaca ggtaggcgcg ccgaagttcc tatactttct agagaatagg aacttcggaa
     1201 taggaactaa ggaggatatt catatggacc atggctaatt cccatgtcag ccgttaagtg
     1261 ttcctgtgtc actcaaaatt gctttgagag gctctaaggg cttctcagtg cgttacatcc
     1321 ctggcttgtt gtccacaacc gttaaacctt aaaagcttta aaagccttat atattctttt
     1381 ttttcttata aaacttaaaa ccttagaggc tatttaagtt gctgatttat attaatttta
     1441 ttgttcaaac atgagagctt agtacgtgaa acatgagagc ttagtacgtt agccatgaga
     1501 gcttagtacg ttagccatga gggtttagtt cgttaaacat gagagcttag tacgttaaac
     1561 atgagagctt agtacgtgaa acatgagagc ttagtacgta ctatcaacag gttgaactgc
     1621 tgatcttcag atcctctacg ccggacgcat cgtggccgga tcttgcggcc gcaaaaatta
     1681 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1741 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1801 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1861 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1921 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1981 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2041 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2101 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2161 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2221 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2281 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2341 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2401 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2461 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2521 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2581 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2641 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2701 cacatttccc cgaaaagtgc cacctgcatc gatggccccc gatggtagtg tggggtctcc
     2761 ccatgcgaga gtagggaact gccaggcatc aaataaaatg aaaggctcag tcgaaagact
     2821 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     2881 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     2941 cataaa
//
