LOCUS       sequence-477853-        5748 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 5748)
  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 5748)
  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: pRE112
FEATURES             Location/Qualifiers
     source          1..5748
                     /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(3221..3880)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIHSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3795..3814
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3881..3983)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     oriT            4712..4821
                     /note="incP origin of transfer"
     CDS             4854..5225
                     /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 atcgggccct atcacttatt caggcgtagc
     3181 aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact
     3241 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     3301 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     3361 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     3421 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     3481 aataggccag gttttcaccg taacacgcca catcttgcga atgtatgtgt agaaactgcc
     3541 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     3601 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     3661 ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     3721 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     3781 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     3841 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     3901 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     3961 tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc
     4021 cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta
     4081 ttagggcccg atcctttttg tccggtgttg ggttgaaggt gaagccggtc ggggccgcag
     4141 cgggggccgg cttttcagcc ttgcccccct gcttcggccg ccgtggctcc ggcgtcttgg
     4201 gtgccggcgc gggttccgca gccttggcct gcggtgcggg cacatcggcg ggcttggcct
     4261 tgatgtgccg cctggcgtgc gagcggaacg tctcgtagga gaacttgacc ttccccgttt
     4321 cccgcatgtg ctcccaaatg gtgacgagcg catagccgga cgctaacgcc gcctcgacat
     4381 ccgccctcac cgccaggaac gcaaccgcag cctcatcacg ccggcgcttc ttggccgcgc
     4441 gggattcaac ccactcggcc agctcgtcgg tgtagctctt tggcatcgtc tctcgcctgt
     4501 cccctcagtt cagtaatttc ctgcatttgc ctgtttccag tcggtagata ttccacaaaa
     4561 cagcagggaa gcagcgcttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
     4621 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
     4681 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
     4741 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
     4801 ctgctctgcg aggctggccg gctaccgccg gcgtaacaga tgagggcaag cggatggctg
     4861 atgaaaccaa gccaaccagg aagggcagcc cacctatcaa ggtgtactgc cttccagacg
     4921 aacgaagagc gattgaggaa aaggcggcgg cggccggcat gagcctgtcg gcctacctgc
     4981 tggccgtcgg ccagggctac aaaatcacgg gcgtcgtgga ctatgagcac gtccgcgagc
     5041 tggcccgcat caatggcgac ctgggccgcc tgggcggcct gctgaaactc tggctcaccg
     5101 acgacccgcg cacggcgcgg ttcggtgatg ccacgatcct cgccctgctg gcgaagatcg
     5161 aagagaagca ggacgagctt ggcaaggtca tgatgggcgt ggtccgcccg agggcagagc
     5221 catgactttt ttagccgcta aaacggccgg ggggtgcgcg tgattgccaa gcacgtcccc
     5281 atgcgctcca tcaagaagag cgacttcgcg gagctggtga agtacatcac cgacgagcaa
     5341 ggcaagaccg agcgcctggg tcacgtgcgc gtcacgaact gcgaggcaaa caccctgccc
     5401 gctgtcatgg ccgaggtgat ggcgacccag cacggcaaca cccgttccga ggccgacaag
     5461 acctatcacc tgctggttag cttccgcgcg ggagagaagc ccgacgcgga gacgttgcgc
     5521 gcgattgagg accgcatctg cgctgggctt ggcttcgccg agcatcagcg cgtcagtgcc
     5581 gtgcatcacg acaccgacaa cctgcacatc catatcgcca tcaacaagat tcacccgacc
     5641 cgaaacacca tccatgagcc gtatcgggcc taccgcgccc tcgctgacct ctgcgcgacg
     5701 ctcgaacggg actacgggct tgagcgtgac aatcacgaaa cgcggcag
//
