LOCUS       sequence-486154-        6133 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Constitutive expression Promoter (RpoD sigma factor, BT_RS06635), 
            integrates via IntN1; N-/C-terminal mScarlet-I fusion, 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 6133)
  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 6133)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 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: pMS08
FEATURES             Location/Qualifiers
     source          1..6133
                     /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             1733..1798
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(4863..4931,4932..5723)
                     /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(5081..5100)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5854..5963
                     /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 agttgatatc agtgatctgg aagaagcaat
      601 gaaagctgct gttaagtctc cgaatcaggt attgttcctg acaggtgtat tcccatccgg
      661 taaacgcgga tactttgcag ttgatctgac tcaggaataa attataaatt aaggtaagaa
      721 gattgtagga taagctaatg aaatagaaaa aggatgccgt cacacaactt gtcggcattc
      781 ttttttgttt tattagttga aaatatagtg aaaaagttgc ctaaatatgt atgttaacaa
      841 attatttgtc gtaactttgc actccaaatc tgtttttaaa gacccgggca tgccaggatc
      901 cgaattcgag ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc
      961 catatggcag atctcaattg gatatcggcc ggccacgcga tcgctgacgt cggtacccat
     1021 gagtaaagga gaagctgtga ttaaagagtt catgcgcttc aaagttcaca tggagggttc
     1081 tatgaacggt cacgagttcg agatcgaagg cgaaggcgag ggccgtccgt atgaaggcac
     1141 ccagaccgcc aaactgaaag tgactaaagg cggcccgctg cctttttcct gggacatcct
     1201 gagcccgcaa tttatgtacg gttctagggc gttcatcaaa cacccagcgg atatcccgga
     1261 ctattataag cagtcttttc cggaaggttt caagtgggaa cgcgtaatga attttgaaga
     1321 tggtggtgcc gtgaccgtca ctcaggacac ctccctggag gatggcaccc tgatctataa
     1381 agttaaactg cgtggtacta attttccacc tgatggcccg gtgatgcaga aaaagacgat
     1441 gggttgggag gcgtctaccg aacgcttgta tccggaagat ggtgtgctga aaggcgacat
     1501 taaaatggcc ctgcgcctga aagatggcgg ccgctatctg gctgacttca aaaccacgta
     1561 caaagccaag aaacctgtgc agatgcctgg cgcgtacaat gtggaccgca aactggacat
     1621 cacctctcat aatgaagatt atacggtggt agagcaatat gagcgctccg agggtcgtca
     1681 ttctaccggt ggcatggatg aactatacaa aacatgtggt catgacacta gtgactacaa
     1741 ggaccacgac ggcgactata aggaccacga catcgactac aaggatgacg acgacaagtg
     1801 agggccctat aatttggtga attggagtta ttttcatatt tttgcatccc gaagagtttc
     1861 tcttaaagag agaaacatct tttgcatacc ttttccgacc gaatttttat gtcgtaaaga
     1921 ggggctttgc agggggtgga ctcagaaaga tgagaataga tgactattgt agttgaaaca
     1981 catagaaagt tgctgatata cagaccgata cgcatatcgg gatgaaccat gagtacgttc
     2041 ttttctcaaa aaacataaat attcgaaaag agatgcaata aattaaggag aggttatagc
     2101 tgagcatgac aaaaaagaaa ttgcccgttc gttttacggg tcagcacttt actattgata
     2161 aagtgctaat aaaagatgca ataagacaag caaatataag taatcaggat acggttttag
     2221 atattggggc aggcaagggg tttcttactg ttcatttatt aaaaatcgcc aacaatgttg
     2281 ttgctattga aaacgacaca gctttggttg aacatttacg aaaattattt tctgatgccc
     2341 gaaatgttca agttgtcggt tgtgatttta ggaattttgc agttccgaaa tttcctttca
     2401 aagtggtgtc aaatattcct tatggcatta cttccgatat tttcaaaatc ctgatgtttg
     2461 agagtcttgg aaattttctg ggaggttcca ttgtccttca attagaacct acacaaaagt
     2521 tattttcgag gaagctttac aatccatata ccgttttcta tcatactttt tttgatttga
     2581 aacttgtcta tgaggtaggt cctgaaagtt tcttgccacc gccaactgtc aaatcagccc
     2641 tgttaaacat taaaagaaaa cacttatttt ttgattttaa gtttaaagcc aaatacttag
     2701 catttatttc ctatctgtta gagaaacctg atttatctgt aaaaacagct ttaaagtcga
     2761 ttttcaggaa aagtcaggtc aggtcaattt cggaaaaatt cggtttaaac cttaatgctc
     2821 aaattgtttg tttgtctcca agtcaatggt taaactgttt tttggaaatg ctggaagttg
     2881 tccctgaaaa atttcatcct tcgtaggcat gcctcatcgg attcttttaa tatcgctact
     2941 ctctacgttc aaccaaaaga aataatgact tactgctata ttttttgcac gtgtggggaa
     3001 aatgtgggga aaattcaagc aaaagaaaaa gctaagtatt gaactatcaa atacttagct
     3061 ttctttcttg tacccagacc ccgcatttga aataattaaa gtggggaaaa tgtgggtaaa
     3121 aagaaaaatg cggaaaaacg ccacaattac actgtatttc aatatgttat aatcctatta
     3181 aattttaatc caagtttaat cgaattgcaa aatatttagc agatgtgggg aaaatgctgg
     3241 ggaaaatatt tatatttgca gcagagtaaa attcaacctg aaagtctatg aaagtaacct
     3301 ttatcattaa aaaagcagcc aaacgatatg atacagaatc catggctaca atctatgtcc
     3361 gttttagaaa cggaaggcag ttagactccg ttgctcctac tcagttagcc atcaatccca
     3421 atctatggga tgataaagac gaatgtgtaa aaacgaaagc tgtctgcaat gaagaaatgc
     3481 gtacccatat aaatgaagag atacgccagt tgaaaaccta tatcgagaag gtatatcaac
     3541 aagaaaagga agcaatagac aaagaatggc taaaaacaac acttgataaa ttttaccatc
     3601 ctgaaaaata ttttttgccg gaggaagtgg ttatcaagcc taccattgga gaactattcg
     3661 atgaatttct aaacaagcac cctttgtcgg aagtacgaaa gaaaaatttc cgggttgtca
     3721 aaagagcctt actgcgttat gaactatatg taagggctac aaagagagga caaaagggct
     3781 ttatccttga tgtggatttg gtaacacctg acacgcttcg ggatatgtgg gatttctttc
     3841 agaacgaata ccagtattat gaactttacc cgagcattta tgaagccatt cccgaaaaga
     3901 ggacaccaca gcccagaagc aaaaacacgc tgatagactg tttttcaaga atacgcacat
     3961 tcttcctgtg gtgcttcgat aacaaacgca ccacaaacag acctttcgac aagtttccga
     4021 tagaggagtg tacatatggt acaccttatt atataacact cgaagaaagg gacaggattt
     4081 ttaatgcaga cctttctgcc accccacaac tggcaataca gagggatata ttcatatttc
     4141 agacactgat aggatgcagg gtgagcgacc tgtaccgaat gaccaaacta aatgtggtca
     4201 atgaagccat agaatatatt cccaagaaaa ccaaagaggg gaatccggtt acggtacgtg
     4261 ttccacttaa cgacaaagcg aaagaaatcc ttgaacgcta caaagaatat gagggaaaac
     4321 tgttgccgtt catatccgag caaaagtaca atgatgccat aaaaaagata ttcaaattag
     4381 ctggagttga ccgcatcgta acaatcttag acccgttgac gcacaacgaa atcaaacgac
     4441 ctatttatga agtggcaagc agccatctgg caagacgtac gtttatcggc aatatctata
     4501 aaaaagtgaa agacccgaac cttgtttccg cactgtcggg acacaaggag ggaagcaaag
     4561 ctttcagacg atacagggat attgacgaag aaatgaagaa agaccttgta aaactactgg
     4621 actgataaaa gtatcaaaca gtttcataga tagttaggct agcatatagc gactttttta
     4681 caaaatgcta aaatttgttg atttccgtca agcaattgtt gagcaaaaat gtcttttacg
     4741 ataaaatgat acctcaatat caactgttta gcaaaacgat atttctctta aagagagaaa
     4801 cacctttttg ttcaccaatc cccgactttt aatcccgcgg ccatgattga aaaaggaaga
     4861 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4921 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4981 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5041 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5101 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5161 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5221 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5281 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5341 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5401 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5461 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5521 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5581 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5641 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5701 cctcactgat taagcattgg taactgtcag accaagttta ctcataacgc gtcaattcga
     5761 gggggatcaa ttccgtgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     5821 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     5881 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     5941 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     6001 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     6061 ctataatgac cccgaagcag ggttatgcag cggaaaacgg aattgatccg gccacgatgc
     6121 gtccggcgta gag
//
