LOCUS       sequence-313593-        4366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gentamycin version of the scarless genome engineering pDEL plasmid 
            (Tikh et al. 2016) compatible with Salmonella.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4366)
  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 4366)
  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: pOD_004
FEATURES             Location/Qualifiers
     source          1..4366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        532..650
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     promoter        1626..1728
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1729..2262
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    2579..2600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2615..2620,2621..2638,2639..2645)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2653..2669
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2658..2680
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(2739..2825,2826..4160)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 catggctaac gtactaagct ctcatgtttc acgtactaag ctctcatgtt tgaacaataa
       61 aattaatata aatcagcaac ttaaatagcc tctaaggttt taagttttat aagaaaaaaa
      121 agaatatata aggcttttaa agctagcttt taaggtttaa cggttgtgga caacaagcca
      181 gggatgtaac gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga
      241 acacttaacg gctgacatgg ccttagaagg cgttttagaa gccctaatgg atggagtcaa
      301 aacaaactag cgatctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      361 tgtaattctc catggtaatt cacgacggag gccacctcat gttagtcatg cgcaaaaaac
      421 cccgcttcgg cggggttttt tcgcgcccgc cgaaaacccc gcttcggcgg ggttttgccg
      481 ctcagctact agggataaca gggtaatgac caggtccttc agaacgttca gcaccacaat
      541 tcagcaaatt gtgaacatca tcacgttcat ctttccctgg ttgccaatgg cccattttcc
      601 tgtcagtaac gagaaggtcg cgaattcagg cgctttttag actggtcgta gtttaacttt
      661 aaggagggga tctatgaaga acatcaaaaa gaatcaagta atcaatttcg gacctaattc
      721 taaattatta aaagaatata aatcacaatt aattgaatta aatattgaac aatttgaagc
      781 aggtattggt ttaattttag gagatgctta tattcgtagt cgtgatgaag gtaaactata
      841 ttgtatgcaa tttgagtgga aaaataaggc atacatggat catgtatgtt tattatatga
      901 tcaatgggta ttatcacctc ctcataaaaa agaaagagtt aatcatttag gtaatttagt
      961 aattacctgg ggagctcaaa ctttcaaaca tcaagctttt aataaattgg ctaacttatt
     1021 cattgtaaat aacaaaaagc ttattcctaa caatttagtt gaaaattact tgacacctat
     1081 aagtttggca tactggttta tagatgatgg aggtaaatgg gattacaata aaaactctct
     1141 taataaaagt atcgtattaa atacacaaag ttttactttc gaagaagtag aatatttggt
     1201 taaaggttta agaaataaat tccaattgaa ttgttatgtt aaaatcaata aaaacaaacc
     1261 aatcatctac attgattcta taagttattt aatcttttat aatttgatca aaccttactt
     1321 aatccctcaa atgatataca aattacctaa tactatctca tccgaaactt tcttgaaata
     1381 acatggcgat agctagactg ggcggtttta tggacagcaa gccaccaaac accccccaaa
     1441 accaagccac tggagcacct caaaaacacc atcatacact aaatcagtaa gttggcagca
     1501 ccacccgacg cactttgcgc cgaataaata cctgtgacgg aagatcactt cgcagaataa
     1561 ataaatcctg gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga
     1621 gacgttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     1681 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat gttacgcagc
     1741 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg
     1801 ggcatcattc gcacatgtag gctcggccct gaccaagtca aatccatgcg ggctgctctt
     1861 gatcttttcg gtcgtgagtt cggagacgta gccacctact cccaacatca gccggactcc
     1921 gattacctcg ggaacttgct ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa
     1981 gaagcggttg ttggcgctct cgcggcttac gttctgccca agtttgagca gccgcgtagt
     2041 gagatctata tctatgatct cgcagtctcc ggcgagcacc ggaggcaggg cattgccacc
     2101 gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg
     2161 caagcagatt acggtgacga tcccgcagtg gctctctata caaagttggg catacgggaa
     2221 gaagtgatgc actttgatat cgacccaagt accgccacct aattttttta aggcagttat
     2281 tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg ataataagcg gatgaatggc
     2341 agaaattcga aagcaaattc gacccggtcg tcggttcagg gcagggtcgt taaatagccg
     2401 cttatgtcta ttgctggttt accggtttat tgactaccgg aagcagtgtg accgtgtgct
     2461 tctcaaatgc ctgaggccag tttgctcagg ctctcccgtg tggtgtggtg tggttgtgtg
     2521 gggttcgaaa tgaccgacca agcgacatca taacggttct ggcaaatatt ctggcaatta
     2581 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2641 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagccag tccgtttagg
     2701 tgttttcacg agcaattgac caacaggagg atctggaaat gaacatcaaa aagtttgcaa
     2761 aacaagcaac agtattaacc tttactaccg cactgctggc aggaggcgca actcaagcgt
     2821 ttgcgaaaga aacgaaccaa aagccatata aggaaacata cggcatttcc catattacac
     2881 gccatgatat gctgcaaatc cctgaacagc aaaaaaatga aaaatatcaa gttcctgagt
     2941 tcgattcgtc cacaattaaa aatatctctt ctgcaaaagg cctggacgtt tgggacagct
     3001 ggccattaca aaacgctgac ggcactgtcg caaactatca cggctaccac atcgtctttg
     3061 cattagccgg agatcctaaa aatgcggatg acacatcgat ttacatgttc tatcaaaaag
     3121 tcggcgaaac ttctattgac agctggaaaa acgctggccg cgtctttaaa gacagcgaca
     3181 aattcgatgc aaatgattct atcctaaaag accaaacaca agaatggtca ggttcagcca
     3241 catttacatc tgacggaaaa atccgtttat tctacactga tttctccggt aaacattacg
     3301 gcaaacaaac actgacaact gcacaagtta acgtatcagc atcagacagc tctttgaaca
     3361 tcaacggtgt agaggattat aaatcaatct ttgacggtga cggaaaaacg tatcaaaatg
     3421 tacagcagtt catcgatgaa ggcaactaca gctcaggcga caaccatacg ctgagagatc
     3481 ctcactacgt agaagataaa ggccacaaat acttagtatt tgaagcaaac actggaactg
     3541 aagatggcta ccaaggcgaa gaatctttat ttaacaaagc atactatggc aaaagcacat
     3601 cattcttccg tcaagaaagt caaaaacttc tgcaaagcga taaaaaacgc acggctgagt
     3661 tagcaaacgg cgctctcggt atgattgagc taaacgatga ttacacactg aaaaaagtga
     3721 tgaaaccgct gattgcatct aacacagtaa cagatgaaat tgaacgcgcg aacgtcttta
     3781 aaatgaacgg caaatggtat ctgttcactg actcccgcgg atcaaaaatg acgattgacg
     3841 gcattacgtc taacgatatt tacatgcttg gttatgtttc taattcttta actggcccat
     3901 acaagccgct gaacaaaact ggccttgtgt taaaaatgga tcttgatcct aacgatgtaa
     3961 cctttactta ctcacacttc gctgtacctc aagcgaaagg aaacaatgtc gtgattacaa
     4021 gctatatgac aaacagagga ttctacgcag acaaacaatc aacgtttgcg cccagcttcc
     4081 tgctgaacat caaaggcaag aaaacatctg ttgtcaaaga cagcatcctt gaacaaggac
     4141 aattaacagt taacaaataa cgtttaaggg caccaataac tgcagcgcta gtacggttat
     4201 ccacagaatc aggggataac gcaggaaaga acatgtgaga tcagcagttc aacctgttga
     4261 tagtacgtac taagctctca tgtttcacgt actaagctct catgtttaac gtactaagct
     4321 ctcatgttta acgaactaaa ccctcatggc taacgtacta agctct
//
