LOCUS       sequence-486619-        6048 bp    DNA     circular SYN 17-SEP-2025
DEFINITION  Inducible Promoter (rhamnose-induced, BT3763/rhaR system), 
            replicates via RepA; ermF, C-terminal 3x-FLAG, R6K ori, RP4 oriT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6048)
  AUTHORS   Schnizlein MK, Dubey AA, Fiebig A, Crosson S
  TITLE     Genetic- and culture-based tools for studying Bacteroides fragilis.
  JOURNAL   Microbiol Resour Announc. 2025 May 8;14(5):e0000625. doi: 
            10.1128/mra.00006-25. Epub 2025 Mar 25.
  PUBMED    40130927
REFERENCE   2  (bases 1 to 6048)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microbiol 
            Resour Announc."; date: "2025-05-8"; volume: "14(5):e0000625. doi"; 
            pages: " 10.1128/mra.00006-25. Epub 2025 Mar 25"
COMMENT     Sequence Label: pMS07
FEATURES             Location/Qualifiers
     source          1..6048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..389
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             1759..1824
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(4778..4846,4847..5638)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4996..5015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5769..5878
                     /note="incP origin of transfer"
ORIGIN
        1 gatctgaaga tcagcagttc aacctgttga tagtacgtac taagctctca tgtttcacgt
       61 actaagctct catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc
      121 taacgtacta agctctcatg gctaacgtac taagctctca tgtttcacgt actaagctct
      181 catgtttgaa caataaaatt aatataaatc agcaacttaa atagcctcta aggttttaag
      241 ttttataaga aaaaaaagaa tatataaggc ttttaaagct tttaaggttt aacggttgtg
      301 gacaacaagc cagggatgta acgcactgag aagcccttag agcctctcaa agcaattttg
      361 agtgacacag gaacacttaa cggctgacat gggaattccc ctccacctta ttcttcccga
      421 ccgaacgagc ggtattcaag cggagtaagc ccggtacgtt ttttgaaaaa ggagaaaaaa
      481 tattcggtag actggaagcc cagaaaaaaa gaaatttctt ttactgactg ggaagttccc
      541 accaacattt gcttggcttt acgaagtttc agttcctgaa aatatttggc gggtgcatag
      601 ccggtatatt ccttaaaaac cctgcggaac catgagtaac tgatattcag tctcatcgcc
      661 agttcctccg gatcaatatt gccggataca ttttcattca taagtatctt tgcctgctcg
      721 atcttctgat cgacatcact catctcgaag accttattct tggaaatcga aagaatcata
      781 ccaatcatgt gaagtacgat tccggaaagg tattgctggg cagatatttt atccgcttct
      841 gccacttcca aagcgcggga aaacaaagat accagttctt cattaatacc gacttccaat
      901 atctgtcttt cctgagaaag aaaggcatcg cccacaatcg tatcaatcgc cggaccttca
      961 aaaccaatgt aatattccgt ccatccggtc tgccgcaatg gataataagt atgccactgt
     1021 ccgggaaaca acaccatcag ccgaccttta caaacttgtc tttcaggagt agaatcggaa
     1081 gaaaacaaac cgcgcccttt tgtaatatag accaattgat attcccgcaa tactcttcct
     1141 ttttcggcat tgaaatagta accggacgga tgctccttca gcggataagg agaatcggga
     1201 gggatagact gatagccaac cgtattcacc cacagaccga atttgcggtc catgtcatta
     1261 actatcaagt atttaaattc taatcccaaa ttattatcct cagtcataca ggcgtgtttt
     1321 catacaattg aattgaacaa aagtaggaat tattttcaaa aaacacacgc taaacagtaa
     1381 aaaaacatcc agagcacctg tccagatcct ccggatgctt attatttcgg agaaaacatg
     1441 cataaatcat gctttttttg cataaaaagt aaaatttata ctgatgtaag gtttggctat
     1501 gcagatttgt gtcaaaatgc acatcctttc tatcaaaatg cgtaaggaaa aggaggaagg
     1561 aaccgcctat ctttgcaatg taggtaaatg gataccttaa atatatagac aaaatacccg
     1621 ggcatgccag gatccgaatt cgagctcggc gcgcctgcag gtcgacaagc ttgcggccgc
     1681 ataatgctta agtccatatg gcagatctca attggatatc ggccggccac gcgatcgctg
     1741 acgtcggtac cctcgagtga ctacaaggac cacgacggcg actataagga ccacgacatc
     1801 gactacaagg atgacgacga caagtgaggg ccctataatt tggtgaattg gagttatttt
     1861 catatttttg catcccgaag agtttctctt aaagagagaa acatcttttg catacctttt
     1921 ccgaccgaat ttttatgtcg taaagagggg ctttgcaggg ggtggactca gaaagatgag
     1981 aatagatgac tattgtagtt gaaacacata gaaagttgct gatatacaga ccgatacgca
     2041 tatcgggatg aaccatgagt acgttctttt ctcaaaaaac ataaatattc gaaaagagat
     2101 gcaataaatt aaggagaggt tatagcttag catgacaaaa aagaaattgc ccgttcgttt
     2161 tacgggtcag cactttacta ttgataaagt gctaataaaa gatgcaataa gacaagcaaa
     2221 tataagtaat caggatacgg ttttagatat tggggcaggc aaggggtttc ttactgttca
     2281 tttattaaaa atcgccaaca atgttgttgc tattgaaaac gacacagctt tggttgaaca
     2341 tttacgaaaa ttattttctg atgcccgaaa tgttcaagtt gtcggttgtg attttaggaa
     2401 ttttgcagtt ccgaaatttc ctttcaaagt ggtgtcaaat attccttatg gcattacttc
     2461 cgatattttc aaaatcctga tgtttgagag tcttggaaat tttctgggag gttccattgt
     2521 ccttcaatta gaacctacac aaaagttatt ttcgaggaag ctttacaatc catataccgt
     2581 tttctatcat actttttttg atttgaaact tgtctatgag gtaggtcctg aaagtttctt
     2641 gccaccgcca actgtcaaat cagccctgtt aaacattaaa agaaaacact tattttttga
     2701 ttttaagttt aaagccaaat acttagcatt tatttcctat ctgttagaga aacctgattt
     2761 atctgtaaaa acagctttaa agtcgatttt caggaaaagt caggtcaggt caatttcgga
     2821 aaaattcggt ttaaacctta atgctcaaat tgtttgtttg tctccaagtc aatggttaaa
     2881 ctgttttttg gaaatgctgg aagttgtccc tgaaaaattt catccttcgt aggcatgcgg
     2941 cgctccccga ccgatgattt ttaagtgact gattttgtgc tgttttgggg gtatattaag
     3001 aataaaagaa atagaataag ttaagtactt gatacacaat atagggcatt ttccatattg
     3061 gaaattctca ttttccaatc tagaaaatac cgatttccta atatgatact aaataggaaa
     3121 ataatatttc ccttaatatt gtttttatgg aaaataatag tttactttgt ggagaataat
     3181 atttcccaaa aacatatcaa aatggaaaat aaaaaagcag ttaagttaac cgattttcaa
     3241 aagaacgaag aaaatccttt tatgaaacaa gctatagaag gtattgaaaa tcatgttgtt
     3301 aaaaagtata agagtaatag tggtggcgat aagagagctg tagtagcttt agccgacact
     3361 gaaactggag aagtgtttaa gacttcgttt atccgtcaaa tagaagtaga tgaagaacaa
     3421 ttcactaaat tgtatctttc taactttgct gcattctttg acctatcaca agcagctatt
     3481 cgggtttttg gttactttat gacctgcatg aaacccaaaa atgatttaat cattttcaat
     3541 agaaaaaaat gcctagaata taccaaatac aaaacagaca aagccgttta taaaggactt
     3601 gcagaacttg taaaagctga aatcatagcc cgaggaccag ccgataatct ttggtttatt
     3661 aatcctctga tagtattcaa tggtgaccga gtgacatttg ctaaaacata cgttcggaaa
     3721 aagactttag ctgcccaaaa gaaagaagaa gcagagaaac gacaattatc acttggcttt
     3781 gatgaacagt aacactccat tgagtgaagc tgccgtttgg tcgctccccc tttgggggcg
     3841 gggggggata gataaagttc ctctatgtaa agttataatg ggggatgaaa ggcaaggttc
     3901 gctaacctta ccccgattgt ttatgcttcc cagcattcaa tctatccccg tttatgggtt
     3961 tctctttcac cctacaaaga taacctcatg ggggaaaaat gtccaagaat atggggtaaa
     4021 aactatcaag tcggtagaaa ataagtatct ttgaagtaca tttttagtag aggtactggt
     4081 gtgcctaggc gaagcagata ggcaaaaata aagccgctgg agatgtcgga tgtccgtcaa
     4141 ttaactccag cggcttgtat tttggtaact ctcccgtatc ggtcagctac cgtttggtca
     4201 gcaacaaagg tagtacttta ttatcgcaaa tccaaatgtt tttcccttta tttgtgcatt
     4261 tacaaaacat tcactaactt cgcacctatg cagtatggtg atagcacact agcaccctaa
     4321 agagtgcgtg tgtaagggga caagccccct taaccccctg tcagccggct gtcaccggct
     4381 gttatcgatt gtcacaaaaa tgttaactta tgggagcaac aagtattcat gtacaagcag
     4441 tgaagccggg gagcgagatt cacaacttta gggaaaaaga gttggactat gttcgtcccg
     4501 aacttagtca tttgaatgaa agctgggttg gagatagcat ttcccatcgg ctggagagtg
     4561 caaaacaaag atgctagcat atagcgactt ttttacaaaa tgctaaaatt tgttgatttc
     4621 cgtcaagcaa ttgttgagca aaaatgtctt ttacgataaa atgatacctc aatatcaact
     4681 gtttagcaaa acgatatttc tcttaaagag agaaacacct ttttgttcac caatccccga
     4741 cttttaatcc cgcggccatg attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4801 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4861 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4921 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4981 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5041 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5101 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5161 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5221 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5281 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5341 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5401 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5461 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5521 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5581 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5641 gtcagaccaa gtttactcat aacgcgtcaa ttcgaggggg atcaattccg tgataggtgg
     5701 gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc
     5761 ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata
     5821 agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg
     5881 ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata
     5941 tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta
     6001 tgcagcggaa aacggaattg atccggccac gatgcgtccg gcgtagag
//
