LOCUS       sequence-476749-        6159 bp    DNA     circular SYN 23-JUN-2025
DEFINITION  Inducible Promoter (rhamnose-induced, BT3763/rhaR system), 
            integrates via IntN1; 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 6159)
  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 6159)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 23, 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: pMS06
FEATURES             Location/Qualifiers
     source          1..6159
                     /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(4889..4957,4958..5749)
                     /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(5107..5126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5880..5989
                     /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 ctcaacctta 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 aggcatgcct
     2941 catcggattc ttttaatatc gctactctct acgttcaacc aaaagaaata atgacttact
     3001 gctatatttt ttgcacgtgt ggggaaaatg tggggaaaat tcaagcaaaa gaaaaagcta
     3061 agtattgaac tatcaaatac ttagctttct ttcttgtacc cagaccccgc atttgaaata
     3121 attaaagtgg ggaaaatgtg ggtaaaaaga aaaatgcgga aaaacgccac aattacactg
     3181 tatttcaata tgttataatc ctattaaatt ttaatccaag tttaatcgaa ttgcaaaata
     3241 tttagcagat gtggggaaaa tgctggggaa aatatttata tttgcagcag agtaaaattc
     3301 aacctgaaag tctatgaaag taacctttat cattaaaaaa gcagccaaac gatatgatac
     3361 agaatccatg gctacaatct atgtccgttt tagaaacgga aggcagttag actccgttgc
     3421 tcctactcag ttagccatca atcccaatct atgggatgat aaagacgaat gtgtaaaaac
     3481 gaaagctgtc tgcaatgaag aaatgcgtac ccatataaat gaagagatac gccagttgaa
     3541 aacctatatc gagaaggtat atcaacaaga aaaggaagca atagacaaag aatggctaaa
     3601 aacaacactt gataaatttt accatcctga aaaatatttt ttgccggagg aagtggttat
     3661 caagcctacc attggagaac tattcgatga atttctaaac aagcaccctt tgtcggaagt
     3721 acgaaagaaa aatttccggg ttgtcaaaag agccttactg cgttatgaac tatatgtaag
     3781 ggctacaaag agaggacaaa agggctttat ccttgatgtg gatttggtaa cacctgacac
     3841 gcttcgggat atgtgggatt tctttcagaa cgaataccag tattatgaac tttacccgag
     3901 catttatgaa gccattcccg aaaagaggac accacagccc agaagcaaaa acacgctgat
     3961 agactgtttt tcaagaatac gcacattctt cctgtggtgc ttcgataaca aacgcaccac
     4021 aaacagacct ttcgacaagt ttccgataga ggagtgtaca tatggtacac cttattatat
     4081 aacactcgaa gaaagggaca ggatttttaa tgcagacctt tctgccaccc cacaactggc
     4141 aatacagagg gatatattca tatttcagac actgatagga tgcagggtga gcgacctgta
     4201 ccgaatgacc aaactaaatg tggtcaatga agccatagaa tatattccca agaaaaccaa
     4261 agaggggaat ccggttacgg tacgtgttcc acttaacgac aaagcgaaag aaatccttga
     4321 acgctacaaa gaatatgagg gaaaactgtt gccgttcata tccgagcaaa agtacaatga
     4381 tgccataaaa aagatattca aattagctgg agttgaccgc atcgtaacaa tcttagaccc
     4441 gttgacgcac aacgaaatca aacgacctat ttatgaagtg gcaagcagcc atctggcaag
     4501 acgtacgttt atcggcaata tctataaaaa agtgaaagac ccgaaccttg tttccgcact
     4561 gtcgggacac aaggagggaa gcaaagcttt cagacgatac agggatattg acgaagaaat
     4621 gaagaaagac cttgtaaaac tactggactg ataaaagtat caaacagttt catagatagt
     4681 taggctagca tatagcgact tttttacaaa atgctaaaat ttgttgattt ccgtcaagca
     4741 attgttgagc aaaaatgtct tttacgataa aatgatacct caatatcaac tgtttagcaa
     4801 aacgatattt ctcttaaaga gagaaacacc tttttgttca ccaatccccg acttttaatc
     4861 ccgcggccat gattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4921 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4981 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5041 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5101 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5161 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5221 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5281 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5341 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5401 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5461 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5521 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5581 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5641 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5701 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5761 agtttactca taacgcgtca attcgagggg gatcaattcc gtgataggtg ggctgccctt
     5821 cctggttggc ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc
     5881 cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggaaagt
     5941 gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg
     6001 ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa
     6061 aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga
     6121 aaacggaatt gatccggcca cgatgcgtcc ggcgtagag
//
