LOCUS       sequence-313592-        3105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gentamycin 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 3105)
  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 3105)
  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_001
FEATURES             Location/Qualifiers
     source          1..3105
                     /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(357..890)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(891..993)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(1246..1279)
                     /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      1328..1716
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(1818..2609,2610..2678))
                     /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     2441..2460
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2679..2783)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2873..2959
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3051..3078
                     /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 cgggagagcc tgagcaaact ggcctcaggc atttgagaag cacacggtca cactgcttcc
      181 ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc
      241 tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca
      301 cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattag
      361 gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta
      421 tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca
      481 agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg
      541 tgctcgccgg agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg
      601 ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg
      661 atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag
      721 taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg
      781 acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt
      841 gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat tttagcttcc
      901 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
      961 tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc
     1021 cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc
     1081 gtcacaggta tttattcggc gcaaagtgcg tcgggtggtg ctgccaactt actgatttag
     1141 tgtatgatgg tgtttttgag gtgctccagt ggcttggttt tgggggtgtt tggtgcagga
     1201 tttatttatt ctgcgaagtg atcttccgtc acaggtaggc gcgccgaagt tcctatactt
     1261 tctagagaat aggaacttcg gaataggaac taaggaggat attcatatgg accatggcta
     1321 attcccatgt cagccgttaa gtgttcctgt gtcactcaaa attgctttga gaggctctaa
     1381 gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac cttaaaagct
     1441 ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga ggctatttaa
     1501 gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt gaaacatgag
     1561 agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta gttcgttaaa
     1621 catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag agcttagtac
     1681 gtactatcaa caggttgaac tgctgatctt cagatcctct acgccggacg catcgtggcc
     1741 ggatcttgcg gccgcaaaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2521 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgc atcgatggcc
     2821 cccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa
     2881 atgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2941 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     3001 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     3061 cctgacggat ggcctttttg cgtggccagt gccaagcttg catgc
//
