LOCUS       sequence-475919-        5773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression Promoter (RpoD sigma factor, BT_RS06635), 
            integrates via IntN1; N-/C-terminal iLOV 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 5773)
  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 5773)
  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: "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: pMS09
FEATURES             Location/Qualifiers
     source          1..5773
                     /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             1373..1438
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(4503..4571,4572..5363)
                     /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(4721..4740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5494..5603
                     /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 gatagagaag aatttcgtca tcactgatcc taggcttccc gataatccca ttatctttgc
     1081 atcagacggc tttcttgaat tgacagagta ttcgcgcgag gaaatattgg ggagaaatgc
     1141 ccggtttctt caggggccag agacagatca agcgactgtc cagaagataa gagacgcaat
     1201 tagagatcag agggagacta ctgtgcagtt gataaactac actaaaagcg gaaagaaatt
     1261 ctggaactta ctccacctgc aacctgtgcg tgatcagaag ggagagcttc aatacttcat
     1321 cggtgtgcag ctcgatggaa gtgatcatgt aacatgtggt catgacacta gtgactacaa
     1381 ggaccacgac ggcgactata aggaccacga catcgactac aaggatgacg acgacaagtg
     1441 agggccctat aatttggtga attggagtta ttttcatatt tttgcatccc gaagagtttc
     1501 tcttaaagag agaaacatct tttgcatacc ttttccgacc gaatttttat gtcgtaaaga
     1561 ggggctttgc agggggtgga ctcagaaaga tgagaataga tgactattgt agttgaaaca
     1621 catagaaagt tgctgatata cagaccgata cgcatatcgg gatgaaccat gagtacgttc
     1681 ttttctcaaa aaacataaat attcgaaaag agatgcaata aattaaggag aggttatagc
     1741 tgagcatgac aaaaaagaaa ttgcccgttc gttttacggg tcagcacttt actattgata
     1801 aagtgctaat aaaagatgca ataagacaag caaatataag taatcaggat acggttttag
     1861 atattggggc aggcaagggg tttcttactg ttcatttatt aaaaatcgcc aacaatgttg
     1921 ttgctattga aaacgacaca gctttggttg aacatttacg aaaattattt tctgatgccc
     1981 gaaatgttca agttgtcggt tgtgatttta ggaattttgc agttccgaaa tttcctttca
     2041 aagtggtgtc aaatattcct tatggcatta cttccgatat tttcaaaatc ctgatgtttg
     2101 agagtcttgg aaattttctg ggaggttcca ttgtccttca attagaacct acacaaaagt
     2161 tattttcgag gaagctttac aatccatata ccgttttcta tcatactttt tttgatttga
     2221 aacttgtcta tgaggtaggt cctgaaagtt tcttgccacc gccaactgtc aaatcagccc
     2281 tgttaaacat taaaagaaaa cacttatttt ttgattttaa gtttaaagcc aaatacttag
     2341 catttatttc ctatctgtta gagaaacctg atttatctgt aaaaacagct ttaaagtcga
     2401 ttttcaggaa aagtcaggtc aggtcaattt cggaaaaatt cggtttaaac cttaatgctc
     2461 aaattgtttg tttgtctcca agtcaatggt taaactgttt tttggaaatg ctggaagttg
     2521 tccctgaaaa atttcatcct tcgtaggcat gcctcatcgg attcttttaa tatcgctact
     2581 ctctacgttc aaccaaaaga aataatgact tactgctata ttttttgcac gtgtggggaa
     2641 aatgtgggga aaattcaagc aaaagaaaaa gctaagtatt gaactatcaa atacttagct
     2701 ttctttcttg tacccagacc ccgcatttga aataattaaa gtggggaaaa tgtgggtaaa
     2761 aagaaaaatg cggaaaaacg ccacaattac actgtatttc aatatgttat aatcctatta
     2821 aattttaatc caagtttaat cgaattgcaa aatatttagc agatgtgggg aaaatgctgg
     2881 ggaaaatatt tatatttgca gcagagtaaa attcaacctg aaagtctatg aaagtaacct
     2941 ttatcattaa aaaagcagcc aaacgatatg atacagaatc catggctaca atctatgtcc
     3001 gttttagaaa cggaaggcag ttagactccg ttgctcctac tcagttagcc atcaatccca
     3061 atctatggga tgataaagac gaatgtgtaa aaacgaaagc tgtctgcaat gaagaaatgc
     3121 gtacccatat aaatgaagag atacgccagt tgaaaaccta tatcgagaag gtatatcaac
     3181 aagaaaagga agcaatagac aaagaatggc taaaaacaac acttgataaa ttttaccatc
     3241 ctgaaaaata ttttttgccg gaggaagtgg ttatcaagcc taccattgga gaactattcg
     3301 atgaatttct aaacaagcac cctttgtcgg aagtacgaaa gaaaaatttc cgggttgtca
     3361 aaagagcctt actgcgttat gaactatatg taagggctac aaagagagga caaaagggct
     3421 ttatccttga tgtggatttg gtaacacctg acacgcttcg ggatatgtgg gatttctttc
     3481 agaacgaata ccagtattat gaactttacc cgagcattta tgaagccatt cccgaaaaga
     3541 ggacaccaca gcccagaagc aaaaacacgc tgatagactg tttttcaaga atacgcacat
     3601 tcttcctgtg gtgcttcgat aacaaacgca ccacaaacag acctttcgac aagtttccga
     3661 tagaggagtg tacatatggt acaccttatt atataacact cgaagaaagg gacaggattt
     3721 ttaatgcaga cctttctgcc accccacaac tggcaataca gagggatata ttcatatttc
     3781 agacactgat aggatgcagg gtgagcgacc tgtaccgaat gaccaaacta aatgtggtca
     3841 atgaagccat agaatatatt cccaagaaaa ccaaagaggg gaatccggtt acggtacgtg
     3901 ttccacttaa cgacaaagcg aaagaaatcc ttgaacgcta caaagaatat gagggaaaac
     3961 tgttgccgtt catatccgag caaaagtaca atgatgccat aaaaaagata ttcaaattag
     4021 ctggagttga ccgcatcgta acaatcttag acccgttgac gcacaacgaa atcaaacgac
     4081 ctatttatga agtggcaagc agccatctgg caagacgtac gtttatcggc aatatctata
     4141 aaaaagtgaa agacccgaac cttgtttccg cactgtcggg acacaaggag ggaagcaaag
     4201 ctttcagacg atacagggat attgacgaag aaatgaagaa agaccttgta aaactactgg
     4261 actgataaaa gtatcaaaca gtttcataga tagttaggct agcatatagc gactttttta
     4321 caaaatgcta aaatttgttg atttccgtca agcaattgtt gagcaaaaat gtcttttacg
     4381 ataaaatgat acctcaatat caactgttta gcaaaacgat atttctctta aagagagaaa
     4441 cacctttttg ttcaccaatc cccgactttt aatcccgcgg ccatgattga aaaaggaaga
     4501 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4561 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4621 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4681 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4741 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4801 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4861 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4921 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4981 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5041 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5101 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5161 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5221 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5281 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5341 cctcactgat taagcattgg taactgtcag accaagttta ctcataacgc gtcaattcga
     5401 gggggatcaa ttccgtgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     5461 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     5521 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     5581 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     5641 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     5701 ctataatgac cccgaagcag ggttatgcag cggaaaacgg aattgatccg gccacgatgc
     5761 gtccggcgta gag
//
