LOCUS       sequence-475920-        6142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression Promoter (RpoD sigma factor, BT_RS06635), 
            integrates via IntN1; N-/C-terminal mNeonGreen 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 6142)
  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 6142)
  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: pMS10
FEATURES             Location/Qualifiers
     source          1..6142
                     /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             1742..1807
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(4872..4940,4941..5732)
                     /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(5090..5109)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5863..5972
                     /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 ccatggtcag
     1021 caaaggtgaa gaagacaata tggcgagcct gccggcgacg catgaactgc acattttcgg
     1081 tagcatcaac ggtgtggact tcgatatggt tggccagggt accggcaacc cgaatgacgg
     1141 ttatgaagaa ctgaatctga aaagtacgaa gggcgatctg caattttccc cgtggattct
     1201 ggtcccgcat atcggttacg gcttccacca gtacctgccg tatccggatg gtatgagccc
     1261 gtttcaagcg gccatggtgg acggttctgg ctatcaggtt catcgtacca tgcaattcga
     1321 agatggcgcg agtctgaccg tgaactatcg ctacacgtat gaaggttcac acattaaagg
     1381 cgaagcgcag gtcaaaggta ccggttttcc ggcggatggt ccggtgatga ccaactcact
     1441 gacggcagct gattggtgcc gttcgaaaaa gacctatccg aatgacaaaa ccattatcag
     1501 cacgtttaag tggtcttaca ccacgggtaa cggcaaacgt tatcgcagca ccgcacgtac
     1561 cacgtacacg ttcgctaaac cgatggcggc caactacctg aaaaatcagc cgatgtatgt
     1621 ttttcgcaaa accgaactga aacattccaa gacggaactg aacttcaagg aatggcaaaa
     1681 ggcgtttacg gatgtgattg gtatggacga actgtacaag acatgtggtc atgacactag
     1741 tgactacaag gaccacgacg gcgactataa ggaccacgac atcgactaca aggatgacga
     1801 cgacaagtga gggccctata atttggtgaa ttggagttat tttcatattt ttgcatcccg
     1861 aagagtttct cttaaagaga gaaacatctt ttgcatacct tttccgaccg aatttttatg
     1921 tcgtaaagag gggctttgca gggggtggac tcagaaagat gagaatagat gactattgta
     1981 gttgaaacac atagaaagtt gctgatatac agaccgatac gcatatcggg atgaaccatg
     2041 agtacgttct tttctcaaaa aacataaata ttcgaaaaga gatgcaataa attaaggaga
     2101 ggttatagct tagcatgaca aaaaagaaat tgcccgttcg ttttacgggt cagcacttta
     2161 ctattgataa agtgctaata aaagatgcaa taagacaagc aaatataagt aatcaggata
     2221 cggttttaga tattggggca ggcaaggggt ttcttactgt tcatttatta aaaatcgcca
     2281 acaatgttgt tgctattgaa aacgacacag ctttggttga acatttacga aaattatttt
     2341 ctgatgcccg aaatgttcaa gttgtcggtt gtgattttag gaattttgca gttccgaaat
     2401 ttcctttcaa agtggtgtca aatattcctt atggcattac ttccgatatt ttcaaaatcc
     2461 tgatgtttga gagtcttgga aattttctgg gaggttccat tgtccttcaa ttagaaccta
     2521 cacaaaagtt attttcgagg aagctttaca atccatatac cgttttctat catacttttt
     2581 ttgatttgaa acttgtctat gaggtaggtc ctgaaagttt cttgccaccg ccaactgtca
     2641 aatcagccct gttaaacatt aaaagaaaac acttattttt tgattttaag tttaaagcca
     2701 aatacttagc atttatttcc tatctgttag agaaacctga tttatctgta aaaacagctt
     2761 taaagtcgat tttcaggaaa agtcaggtca ggtcaatttc ggaaaaattc ggtttaaacc
     2821 ttaatgctca aattgtttgt ttgtctccaa gtcaatggtt aaactgtttt ttggaaatgc
     2881 tggaagttgt ccctgaaaaa tttcatcctt cgtaggcatg cctcatcgga ttcttttaat
     2941 atcgctactc tctacgttca accaaaagaa ataatgactt actgctatat tttttgcacg
     3001 tgtggggaaa atgtggggaa aattcaagca aaagaaaaag ctaagtattg aactatcaaa
     3061 tacttagctt tctttcttgt acccagaccc cgcatttgaa ataattaaag tggggaaaat
     3121 gtgggtaaaa agaaaaatgc ggaaaaacgc cacaattaca ctgtatttca atatgttata
     3181 atcctattaa attttaatcc aagtttaatc gaattgcaaa atatttagca gatgtgggga
     3241 aaatgctggg gaaaatattt atatttgcag cagagtaaaa ttcaacctga aagtctatga
     3301 aagtaacctt tatcattaaa aaagcagcca aacgatatga tacagaatcc atggctacaa
     3361 tctatgtccg ttttagaaac ggaaggcagt tagactccgt tgctcctact cagttagcca
     3421 tcaatcccaa tctatgggat gataaagacg aatgtgtaaa aacgaaagct gtctgcaatg
     3481 aagaaatgcg tacccatata aatgaagaga tacgccagtt gaaaacctat atcgagaagg
     3541 tatatcaaca agaaaaggaa gcaatagaca aagaatggct aaaaacaaca cttgataaat
     3601 tttaccatcc tgaaaaatat tttttgccgg aggaagtggt tatcaagcct accattggag
     3661 aactattcga tgaatttcta aacaagcacc ctttgtcgga agtacgaaag aaaaatttcc
     3721 gggttgtcaa aagagcctta ctgcgttatg aactatatgt aagggctaca aagagaggac
     3781 aaaagggctt tatccttgat gtggatttgg taacacctga cacgcttcgg gatatgtggg
     3841 atttctttca gaacgaatac cggtattatg aactttaccc gagcatttat gaagccattc
     3901 ccgaaaagag gacaccacag cccagaagca aaaacacgct gatagactgt ttttcaagaa
     3961 tacgcacatt cttcctgtgg tgcttcgata acaaacgcac cacaaacaga cctttcgaca
     4021 agtttccgat agaggagtgt acatatggta caccttatta tataacactc gaagaaaggg
     4081 acaggatttt taatgcagac ctttctgcca ccccacaact ggcaatacag agggatatat
     4141 tcatatttca gacactgata ggatgcaggg tgagcgacct gtaccgaatg accaaactaa
     4201 atgtggtcaa tgaagccata gaatatattc ccaagaaaac caaagagggg aatccggtta
     4261 cggtacgtgt tccacttaac gacaaagcga aagaaatcct tgaacgctac aaagaatatg
     4321 agggaaaact gttgccgttc atatccgagc aaaagtacaa tgatgccata aaaaagatat
     4381 tcaaattagc tggagttgac cgcatcgtaa caatcttaga cccgttgacg cacaacgaaa
     4441 tcaaacgacc tatttatgaa gtggcaagca gccatctggc aagacgtacg tttatcggca
     4501 atatctataa aaaagtgaaa gacccgaacc ttgtttccgc actgtcggga cacaaggagg
     4561 gaagcaaagc tttcagacga tacagggata ttgacgaaga aatgaagaaa gaccttgtaa
     4621 aactactgga ctgataaaag tatcaaacag tttcatagat agttaggcta gcatatagcg
     4681 acttttttac aaaatgctaa aatttgttga tttccgtcaa gcaattgttg agcaaaaatg
     4741 tcttttacga taaaatgata cctcaatatc aactgtttag caaaacgata tttctcttaa
     4801 agagagaaac acctttttgt tcaccaatcc ccgactttta atcccgcggc catgattgaa
     4861 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4921 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4981 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5041 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5101 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5161 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5221 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5281 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5341 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5401 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5461 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5521 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5581 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5641 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5701 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcataacgcg
     5761 tcaattcgag ggggatcaat tccgtgatag gtgggctgcc cttcctggtt ggcttggttt
     5821 catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag
     5881 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     5941 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     6001 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     6061 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaacgga attgatccgg
     6121 ccacgatgcg tccggcgtag ag
//
