LOCUS       sequence-434063-        6187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6187)
  AUTHORS   Edwards RA, Keller LH, Schifferli DM
  TITLE     Improved allelic exchange vectors and their use to analyze 987P 
            fimbria gene expression.
  JOURNAL   Gene. 1998 Jan 30;207(2):149-57.
  PUBMED    9511756
REFERENCE   2  (bases 1 to 6187)
  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: "Gene."; 
            date: "1998-01-30"; volume: "207(2)"; pages: "149-57"
COMMENT     Sequence Label: pDMS197
FEATURES             Location/Qualifiers
     source          1..6187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      99..487
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     promoter        515..960
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(961..1047,1048..2382)
                     /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"
     CDS             complement(3251..4441)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     4070..4089
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(4489..4494,4495..4511,4512..4517))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     oriT            5151..5260
                     /note="incP origin of transfer"
     CDS             5293..5664
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 cgcgtttccg agaaccgcgc gaacgacatg gagcggcacg cgggcgtgga aagcctggtc
       61 ggctggatcg atccggccac gatgcgtccg gcgtagagga tctgaagatc agcagttcaa
      121 cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
      181 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc
      241 taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa
      301 tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata
      361 tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac
      421 gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg
      481 gctgacatgg gaattctgat cctttttaac ccatcacata tacctgccgt tcactattat
      541 ttagtgaaat gagatattat gatattttct gaattgtgat taaaaaggca actttatgcc
      601 catgcaacag aaactataaa aaatacagag aatgaaaaga aacagataga ttttttagtt
      661 ctttaggccc gtagtctgca aatcctttta tgattttcta tcaaacaaaa gaggaaaata
      721 gaccagttgc aatccaaacg agagtctaat agaatgaggt cgaaaagtaa atcgcgcggg
      781 tttgttactg ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg tatactttgg
      841 cgtcacccct tacatatttt aggtcttttt ttattgtgcg taactaactt gccatcttca
      901 aacaggaggg ctggaagaag cagaccgcta acacagtaca taaaaaagga gacatgaacg
      961 atgaacatca aaaagtttgc aaaacaagca acagtattaa cctttactac cgcactgctg
     1021 gcaggaggcg caactcaagc gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca
     1081 tacggcattt cccatattac acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat
     1141 gaaaaatatc aagttcctga gttcgattcg tccacaatta aaaatatctc ttctgcaaaa
     1201 ggcctggacg tttgggacag ctggccatta caaaacgctg acggcactgt cgcaaactat
     1261 cacggctacc acatcgtctt tgcattagcc ggagatccta aaaatgcgga tgacacatcg
     1321 atttacatgt tctatcaaaa agtcggcgaa acttctattg acagctggaa aaacgctggc
     1381 cgcgtcttta aagacagcga caaattcgat gcaaatgatt ctatcctaaa agaccaaaca
     1441 caagaatggt caggttcagc cacatttaca tctgacggaa aaatccgttt attctacact
     1501 gatttctccg gtaaacatta cggcaaacaa acactgacaa ctgcacaagt taacgtatca
     1561 gcatcagaca gctctttgaa catcaacggt gtagaggatt ataaatcaat ctttgacggt
     1621 gacggaaaaa cgtatcaaaa tgtacagcag ttcatcgatg aaggcaacta cagctcaggc
     1681 gacaaccata cgctgagaga tcctcactac gtagaagata aaggccacaa atacttagta
     1741 tttgaagcaa acactggaac tgaagatggc taccaaggcg aagaatcttt atttaacaaa
     1801 gcatactatg gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc
     1861 gataaaaaac gcacggctga gttagcaaac ggcgctctcg gtatgattga gctaaacgat
     1921 gattacacac tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt aacagatgaa
     1981 attgaacgcg cgaacgtctt taaaatgaac ggcaaatggt atctgttcac tgactcccgc
     2041 ggatcaaaaa tgacgattga cggcattacg tctaacgata tttacatgct tggttatgtt
     2101 tctaattctt taactggccc atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg
     2161 gatcttgatc ctaacgatgt aacctttact tactcacact tcgctgtacc tcaagcgaaa
     2221 ggaaacaatg tcgtgattac aagctatatg acaaacagag gattctacgc agacaaacaa
     2281 tcaacgtttg cgcccagctt cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa
     2341 gacagcatcc ttgaacaagg acaattaaca gttaacaaat aaaaacgcaa aagaaaatgc
     2401 cgatatccta ttggcatttt cttttatttc ttatcaacat aaaggtgaat cccatatgaa
     2461 ctatataaaa gcaggcaaat ggctaaccgt attcctaacc ttttggtaat gactccaact
     2521 tattgatagt gttttatgtt cagataatgc ccgatgactt tgtcatgcag ctccaccgat
     2581 tttgagaacg acagcgactt ccgtcccagc cgtgccaggt gctgcctcag attcaggtta
     2641 tgccgctcaa ttcgctgcgt atatcgcttg ctgattacgt gcagctttcc cttcaggcgg
     2701 gattcataca gcggccagcc atccgtcatc catatcacca cgtcaaaggg tgacagcagg
     2761 ctcataagac gccccagcgt cgccatagtg cgttcaccga atacgtgcgc aacaaccgtc
     2821 ttccggagac tgtcatacgc gtaaaacagc cagcgctggc gcgatttagc cccgacatag
     2881 ccccactgtt cgtccatttc cgcgcagacg atgacgtcac tgcccggctg tatgcgcgag
     2941 gttaccgact gcggcctgag ttttttaagt gacgtaaaat cgtgttgagg ccaacgccca
     3001 taatgcgggc tgttgcccgg catccaacgc cattcatggc catatcaatg attttctggt
     3061 gcgtaccggg ttgagaagcg gtgtaagtga actgcatgaa ttcccgggag agctcgatat
     3121 cgcatgcggt acctctagaa gaagcttggg atccaagggt tggtttgcgc attcacagtt
     3181 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     3241 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     3301 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     3361 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     3421 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     3481 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     3541 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     3601 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     3661 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     3721 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     3781 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     3841 tgactgggtt gaaggctctc aagggcatcg gtcgacgctc tcccttatgc gactcctgca
     3901 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     3961 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     4021 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     4081 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     4141 cgtagaggat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata gtggctccaa
     4201 gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct ccgagaacgg
     4261 gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag tgactggcga
     4321 tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata accaagccta
     4381 tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca
     4441 tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat
     4501 cgatgataag ctgtcaaaca tgagaattga tcctttttgt ccggtgttgg gttgaaggtg
     4561 aagccggtcg gggccgcagc gggggccggc ttttcagcct tgcccccctg cttcggccgc
     4621 cgtggctccg gcgtcttggg tgccggcgcg ggttccgcag ccttggcctg cggtgcgggc
     4681 acatcggcgg gcttggcctt gatgtgccgc ctggcgtgcg agcggaacgt ctcgtaggag
     4741 aacttgacct tccccgtttc ccgcatgtgc tcccaaatgg tgacgagcgc atagccggac
     4801 gctaacgccg cctcgacatc cgccctcacc gccaggaacg caaccgcagc ctcatcacgc
     4861 cggcgcttct tggccgcgcg ggattcaacc cactcggcca gctcgtcggt gtagctcttt
     4921 ggcatcgtct ctcgcctgtc ccctcagttc agtaatttcc tgcatttgcc tgtttccagt
     4981 cggtagatat tccacaaaac agcagggaag cagcgctttt ccgctgcata accctgcttc
     5041 ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt
     5101 tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata
     5161 ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg
     5221 gcggtgctca acgggaatcc tgctctgcga ggctggccgg ctaccgccgg cgtaacagat
     5281 gagggcaagc ggatggctga tgaaaccaag ccaaccagga agggcagccc acctatcaag
     5341 gtgtactgcc ttccagacga acgaagagcg attgaggaaa aggcggcggc ggccggcatg
     5401 agcctgtcgg cctacctgct ggccgtcggc cagggctaca aaatcacggg cgtcgtggac
     5461 tatgagcacg tccgcgagct ggcccgcatc aatggcgacc tgggccgcct gggcggcctg
     5521 ctgaaactct ggctcaccga cgacccgcgc acggcgcggt tcggtgatgc cacgatcctc
     5581 gccctgctgg cgaagatcga agagaagcag gacgagcttg gcaaggtcat gatgggcgtg
     5641 gtccgcccga gggcagagcc atgacttttt tagccgctaa aacggccggg gggtgcgcgt
     5701 gattgccaag cacgtcccca tgcgctccat caagaagagc gacttcgcgg agctggtgaa
     5761 gtacatcacc gacgagcaag gcaagaccga gcgcctgggt cacgtgcgcg tcacgaactg
     5821 cgaggcaaac accctgcccg ctgtcatggc cgaggtgatg gcgacccagc acggcaacac
     5881 ccgttccgag gccgacaaga cctatcacct gctggttagc ttccgcgcgg gagagaagcc
     5941 cgacgcggag acgttgcgcg cgattgagga ccgcatctgc gctgggcttg gcttcgccga
     6001 gcatcagcgc gtcagtgccg tgcatcacga caccgacaac ctgcacatcc atatcgccat
     6061 caacaagatt cacccgaccc gaaacaccat ccatgagccg tatcgggcct accgcgccct
     6121 cgctgacctc tgcgcgacgc tcgaacggga ctacgggctt gagcgtgaca atcacgaaac
     6181 gcggcag
//
