LOCUS       sequence-434062-        6065 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 6065)
  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 6065)
  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: pRE118
FEATURES             Location/Qualifiers
     source          1..6065
                     /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"
     primer_bind     complement(3273..3292)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3284..4099
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3284..4099
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3357..3376)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     oriT            5029..5138
                     /note="incP origin of transfer"
     CDS             5171..5542
                     /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 atcgacctgc aggggggggg gggaaagcca
     3181 cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     3241 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     3301 ggaaacgtct tgctcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     3361 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     3421 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3481 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3541 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc
     3601 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3661 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3721 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3781 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt
     3841 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3901 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3961 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     4021 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     4081 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     4141 gctgacttga cgggacggcg gctttgttga ataaatcgaa cttttgctga gttgaaggat
     4201 cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc
     4261 accaactggt ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg
     4321 gatgatgggg cgattcaggc ctggtatgag tcagcaacac cttcttcacg aggcagacct
     4381 cagcgccccc cccccccctg caggtcgatc ctttttgtcc ggtgttgggt tgaaggtgaa
     4441 gccggtcggg gccgcagcgg gggccggctt ttcagccttg cccccctgct tcggccgccg
     4501 tggctccggc gtcttgggtg ccggcgcggg ttccgcagcc ttggcctgcg gtgcgggcac
     4561 atcggcgggc ttggccttga tgtgccgcct ggcgtgcgag cggaacgtct cgtaggagaa
     4621 cttgaccttc cccgtttccc gcatgtgctc ccaaatggtg acgagcgcat agccggacgc
     4681 taacgccgcc tcgacatccg ccctcaccgc caggaacgca accgcagcct catcacgccg
     4741 gcgcttcttg gccgcgcggg attcaaccca ctcggccagc tcgtcggtgt agctctttgg
     4801 catcgtctct cgcctgtccc ctcagttcag taatttcctg catttgcctg tttccagtcg
     4861 gtagatattc cacaaaacag cagggaagca gcgcttttcc gctgcataac cctgcttcgg
     4921 ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg
     4981 ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg
     5041 tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc
     5101 ggtgctcaac gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga
     5161 gggcaagcgg atggctgatg aaaccaagcc aaccaggaag ggcagcccac ctatcaaggt
     5221 gtactgcctt ccagacgaac gaagagcgat tgaggaaaag gcggcggcgg ccggcatgag
     5281 cctgtcggcc tacctgctgg ccgtcggcca gggctacaaa atcacgggcg tcgtggacta
     5341 tgagcacgtc cgcgagctgg cccgcatcaa tggcgacctg ggccgcctgg gcggcctgct
     5401 gaaactctgg ctcaccgacg acccgcgcac ggcgcggttc ggtgatgcca cgatcctcgc
     5461 cctgctggcg aagatcgaag agaagcagga cgagcttggc aaggtcatga tgggcgtggt
     5521 ccgcccgagg gcagagccat gactttttta gccgctaaaa cggccggggg gtgcgcgtga
     5581 ttgccaagca cgtccccatg cgctccatca agaagagcga cttcgcggag ctggtgaagt
     5641 acatcaccga cgagcaaggc aagaccgagc gcctgggtca cgtgcgcgtc acgaactgcg
     5701 aggcaaacac cctgcccgct gtcatggccg aggtgatggc gacccagcac ggcaacaccc
     5761 gttccgaggc cgacaagacc tatcacctgc tggttagctt ccgcgcggga gagaagcccg
     5821 acgcggagac gttgcgcgcg attgaggacc gcatctgcgc tgggcttggc ttcgccgagc
     5881 atcagcgcgt cagtgccgtg catcacgaca ccgacaacct gcacatccat atcgccatca
     5941 acaagattca cccgacccga aacaccatcc atgagccgta tcgggcctac cgcgccctcg
     6001 ctgacctctg cgcgacgctc gaacgggact acgggcttga gcgtgacaat cacgaaacgc
     6061 ggcag
//
