LOCUS       sequence-313590-        4207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zeomycin 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 4207)
  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 4207)
  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_003
FEATURES             Location/Qualifiers
     source          1..4207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        359..477
                     /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        1453..1555
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1556..1930
                     /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"
     protein_bind    2247..2268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2283..2288,2289..2306,2307..2313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2321..2337
                     /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     2326..2348
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(2407..2493,2494..3828)
                     /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 caagccaggg atgtaacgca ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga
       61 cacaggaaca cttaacggct gacatggcct tagaaggcgt tttagaagcc ctaatggatg
      121 gagtcaaaac aaactagcga tctgacgctc agtggaacga aaactcacgt taagggattt
      181 tggtcattgt aattctccat ggtaattcac gacggaggcc acctcatgtt agtcatgcgc
      241 aaaaaacccc gcttcggcgg ggttttttcg cgcccgccga aaaccccgct tcggcggggt
      301 tttgccgctc agctactagg gataacaggg taatgaccag gtccttcaga acgttcagca
      361 ccacaattca gcaaattgtg aacatcatca cgttcatctt tccctggttg ccaatggccc
      421 attttcctgt cagtaacgag aaggtcgcga attcaggcgc tttttagact ggtcgtagtt
      481 taactttaag gaggggatct atgaagaaca tcaaaaagaa tcaagtaatc aatttcggac
      541 ctaattctaa attattaaaa gaatataaat cacaattaat tgaattaaat attgaacaat
      601 ttgaagcagg tattggttta attttaggag atgcttatat tcgtagtcgt gatgaaggta
      661 aactatattg tatgcaattt gagtggaaaa ataaggcata catggatcat gtatgtttat
      721 tatatgatca atgggtatta tcacctcctc ataaaaaaga aagagttaat catttaggta
      781 atttagtaat tacctgggga gctcaaactt tcaaacatca agcttttaat aaattggcta
      841 acttattcat tgtaaataac aaaaagctta ttcctaacaa tttagttgaa aattacttga
      901 cacctataag tttggcatac tggtttatag atgatggagg taaatgggat tacaataaaa
      961 actctcttaa taaaagtatc gtattaaata cacaaagttt tactttcgaa gaagtagaat
     1021 atttggttaa aggtttaaga aataaattcc aattgaattg ttatgttaaa atcaataaaa
     1081 acaaaccaat catctacatt gattctataa gttatttaat cttttataat ttgatcaaac
     1141 cttacttaat ccctcaaatg atatacaaat tacctaatac tatctcatcc gaaactttct
     1201 tgaaataaca tggcgatagc tagactgggc ggttttatgg acagcaagcc accaaacacc
     1261 ccccaaaacc aagccactgg agcacctcaa aaacaccatc atacactaaa tcagtaagtt
     1321 ggcagcacca cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc
     1381 agaataaata aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg
     1441 aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     1501 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatggc
     1561 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgtcggag cggtcgagtt
     1621 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     1681 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggcggtgc cggacaacac
     1741 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     1801 gtccacggac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     1861 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     1921 gcaggactga tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     1981 aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc
     2041 ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac cggtttattg
     2101 actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct
     2161 ctcccgtgtg gtgtggtgtg gttgtgtggg gttcgaaatg accgaccaag cgacatcata
     2221 acggttctgg caaatattct ggcaattaat gtgagttagc tcactcatta ggcaccccag
     2281 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     2341 cacacaggaa acagccagtc cgtttaggtg ttttcacgag caattgacca acaggaggat
     2401 ctggaaatga acatcaaaaa gtttgcaaaa caagcaacag tattaacctt tactaccgca
     2461 ctgctggcag gaggcgcaac tcaagcgttt gcgaaagaaa cgaaccaaaa gccatataag
     2521 gaaacatacg gcatttccca tattacacgc catgatatgc tgcaaatccc tgaacagcaa
     2581 aaaaatgaaa aatatcaagt tcctgagttc gattcgtcca caattaaaaa tatctcttct
     2641 gcaaaaggcc tggacgtttg ggacagctgg ccattacaaa acgctgacgg cactgtcgca
     2701 aactatcacg gctaccacat cgtctttgca ttagccggag atcctaaaaa tgcggatgac
     2761 acatcgattt acatgttcta tcaaaaagtc ggcgaaactt ctattgacag ctggaaaaac
     2821 gctggccgcg tctttaaaga cagcgacaaa ttcgatgcaa atgattctat cctaaaagac
     2881 caaacacaag aatggtcagg ttcagccaca tttacatctg acggaaaaat ccgtttattc
     2941 tacactgatt tctccggtaa acattacggc aaacaaacac tgacaactgc acaagttaac
     3001 gtatcagcat cagacagctc tttgaacatc aacggtgtag aggattataa atcaatcttt
     3061 gacggtgacg gaaaaacgta tcaaaatgta cagcagttca tcgatgaagg caactacagc
     3121 tcaggcgaca accatacgct gagagatcct cactacgtag aagataaagg ccacaaatac
     3181 ttagtatttg aagcaaacac tggaactgaa gatggctacc aaggcgaaga atctttattt
     3241 aacaaagcat actatggcaa aagcacatca ttcttccgtc aagaaagtca aaaacttctg
     3301 caaagcgata aaaaacgcac ggctgagtta gcaaacggcg ctctcggtat gattgagcta
     3361 aacgatgatt acacactgaa aaaagtgatg aaaccgctga ttgcatctaa cacagtaaca
     3421 gatgaaattg aacgcgcgaa cgtctttaaa atgaacggca aatggtatct gttcactgac
     3481 tcccgcggat caaaaatgac gattgacggc attacgtcta acgatattta catgcttggt
     3541 tatgtttcta attctttaac tggcccatac aagccgctga acaaaactgg ccttgtgtta
     3601 aaaatggatc ttgatcctaa cgatgtaacc tttacttact cacacttcgc tgtacctcaa
     3661 gcgaaaggaa acaatgtcgt gattacaagc tatatgacaa acagaggatt ctacgcagac
     3721 aaacaatcaa cgtttgcgcc cagcttcctg ctgaacatca aaggcaagaa aacatctgtt
     3781 gtcaaagaca gcatccttga acaaggacaa ttaacagtta acaaataacg tttaagggca
     3841 ccaataactg cagcgctagt acggttatcc acagaatcag gggataacgc aggaaagaac
     3901 atgtgagatc agcagttcaa cctgttgata gtacgtacta agctctcatg tttcacgtac
     3961 taagctctca tgtttaacgt actaagctct catgtttaac gaactaaacc ctcatggcta
     4021 acgtactaag ctctcatggc taacgtacta agctctcatg tttcacgtac taagctctca
     4081 tgtttgaaca ataaaattaa tataaatcag caacttaaat agcctctaag gttttaagtt
     4141 ttataagaaa aaaaagaata tataaggctt ttaaagctag cttttaaggt ttaacggttg
     4201 tggacaa
//
