LOCUS       sequence-267063-        5040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide vector for allelic replacement in Bacteroides species 
            including B. fragilis strain 638R, erythromycin selection and 
            aTC-inducible ss-Bfe3 counterselection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5040)
  AUTHORS   Ito T, Gallegos R, Matano LM, Butler NL, Hantman N, Kaili M, Coyne 
            MJ, Comstock LE, Malamy MH, Barquera B
  TITLE     Genetic and Biochemical Analysis of Anaerobic Respiration in 
            Bacteroides fragilis and Its Importance In Vivo.
  JOURNAL   mBio. 2020 Feb 4;11(1). pii: mBio.03238-19. doi: 
            10.1128/mBio.03238-19.
  PUBMED    32019804
REFERENCE   2  (bases 1 to 5040)
  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: "mBio."; 
            date: "2020-02-4"; volume: "11(1). pii: mBio.03238-19. doi"; pages: 
            " 10.1128/mBio.03238-19"
COMMENT     Sequence Label: pLGB36
FEATURES             Location/Qualifiers
     source          1..5040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2131..2149
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2193..2211
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2482..3105
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2555..2574)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     rep_origin      3132..3520
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            3691..3800
                     /note="incP origin of transfer"
     CDS             complement(join(3931..4722,4723..4791))
                     /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     4554..4573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcttaaagag agaaacatct tttgcatacc ttttccgacc gaatttttat gtcgtaaaga
       61 ggggctttgc agggggtgga ctcagaaaga tgagaataga tgactattgt agttgaaaca
      121 catagaaagt tgctgatata cagaccgata cgcatatcgg gatgaaccat gagtacgttc
      181 ttttctcaaa aaacataaat attcgaaaag agatgcaata aattaaggag aggttataat
      241 gaacaaagta aatataaaag atagtcaaaa ttttattact tcaaaatatc acatagaaaa
      301 aataatgaat tgcataagtt tagatgaaaa agataacatc tttgaaatag gtgcagggaa
      361 aggtcatttt actgctggat tggtaaagag atgtaatttt gtaacggcga tagaaattga
      421 ttctaaatta tgtgaggtaa ctcgtaataa gctcttaaat tatcctaact atcaaatagt
      481 aaatgatgat atactgaaat ttacatttcc tagccacaat ccatataaaa tatttggcag
      541 cataccttac aacataagca caaatataat tcgaaaaatt gtttttgaaa gttcagccac
      601 aataagttat ttaatagtgg aatatggttt tgctaaaatg ttattagata caaacagatc
      661 actagcattg ctgttaatgg cagaggtaga tatttctata ttagcaaaaa ttcctaggta
      721 ttatttccat ccaaaaccta aagtggatag cacattaatt gtattaaaaa gaaagccagc
      781 aaaaatggca tttaaagaga gaaaaaaata tgaaactttt gtaatgaaat gggttaacaa
      841 agagtacgaa aaactgttta caaaaaatca atttaataaa gctttaaaac atgcgagaat
      901 atatgatata aacaatatta gtttcgaaca atttgtatcg ctatttaata gttataaaat
      961 atttaacggc taaaaacaat aggccacatg caactgttgg attcatacaa gcggtcggtt
     1021 cggaaataga acaggcacaa gtaaggctga ttgttgcagc caatgcacaa atgctgttcc
     1081 atcactggaa gataggcaat tagtcgacgg tatcgataag cttgatatcg aattcctgca
     1141 gcccggggga tccactcatc tgatactatg ccaatgtttc tcatactcat ctaatagctt
     1201 ctcatctcgt ttagccgcat tcttcataat ttcgtcctgt tcttccttgc ttcttccgaa
     1261 atcatttgct atttcataga acttcttcaa actgttgtat tgctcttgtg taatctttgt
     1321 tctgtcaata taccaactgc cgtcttcact ctgttttgca gtgtataatc cgtctctata
     1381 aaatacttcc aaaacacgaa gagccatatc ccgcatacct atcggtgctg tattaccgag
     1441 tgagcgtcct atttgattat tcaacaaatc tattgtctgg tcagcatcga gttgggttct
     1501 gaaccttctg ttgcgaagat tcacatcagg attttcttca tgggcatttc ctatagaagt
     1561 tgctatttct atcccatgcc tattggtgat atagctttgc cacaatgtat ggcggaaagc
     1621 attgctttcc gcgccgtctc cccctgatag tacgtcatta cgggttgcga atcttaccgc
     1681 atttgttgaa atgttggtac ttccttttat ccaataccct atttctgcag ctatcaaagg
     1741 atgggataaa gcgaattttg ccgttctcgc tacagcggat aaacctttag aagaaggttc
     1801 actcgctaaa ggtggaaaac tcaatattat tgtttcttca gaaatgtcca attcctgaac
     1861 ctttattgta tcgaaataat agcaggcagg gtttgcatag agcaaacgtt ttactgtagg
     1921 tgtaataaaa cgaatatagg catatcctac acctataaca aaaccttcat ataatatgcc
     1981 tttgctttta ttatattcaa attctcccgg agagatatat accttcctgt attttttgcc
     2041 ggcaaatgag aactgtatag ctacaatagc catcacaagc aaagataaaa ttttcttcat
     2101 gtgtcttttc ttttatatgt ctttatttcg tctctatcac tgatagggag actgcaaaga
     2161 taagaacttt tagctttatc ttccaaactt tatccctatc agtgatagag atttttttgt
     2221 tcttatggtc aagcagtact gcgcttttac actaagtcaa tttaacaagg ctttcttctc
     2281 ttggaaagcg ggtgcaaaaa aagaaaaggc gttttgtttt tcttctttac cttctttccc
     2341 tttcgctaag agagtctgag aaacgataga aaaagaaaag cgaaaaaact tccgaaaaca
     2401 tttggtagtt aaaataaaac ctcttacctt tgcacccgcg cattttaaaa taaaataaat
     2461 tatttatgat attaaacgaa tatgtctaga ttagataaaa gtaaagtgat taacagcgca
     2521 ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag
     2581 ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac
     2641 gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa
     2701 agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat
     2761 cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc
     2821 gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca
     2881 ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc
     2941 gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct
     3001 atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc
     3061 atatgcggat tagaaaaaca acttaaatgt gaaagtgggt cttaagccac cgcggtggag
     3121 gggaattccc atgtcagccg ttaagtgttc ctgtgtcact caaaattgct ttgagaggct
     3181 ctaagggctt ctcagtgcgt tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa
     3241 agctttaaaa gccttatata ttcttttttt tcttataaaa cttaaaacct tagaggctat
     3301 ttaagttgct gatttatatt aattttattg ttcaaacatg agagcttagt acgtgaaaca
     3361 tgagagctta gtacgttagc catgagagct tagtacgtta gccatgaggg tttagttcgt
     3421 taaacatgag agcttagtac gtgaaacatg agagcttagt acgtgaaaca tgagagctta
     3481 gtacgtacta tcaacaggtt gaactgctga tcttcagatc ctctacgccg gacgcatcgt
     3541 ggccggatca attccgtttt ccgctgcata accctgcttc ggggtcatta tagcgatttt
     3601 ttcggtatat ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga
     3661 ctttccttgg tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc
     3721 gagcgggtgt tccttcttca ctgtccctta ttcgcacctg gcggtgctca acgggaatcc
     3781 tgctctgcga ggctggccgg ctaccgccgg cgtaacagat gagggcaagc ggatggctga
     3841 tgaaaccaag ccaaccagga agggcagccc acctatcacg gaattgatcc ccctcgaatt
     3901 gacgcgttat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4801 ttttcaatca tggccgcggg attaaaagtc ggggattggt gaacaaaaag gtgtttctct
     4861 ctttaagaga aatatcgttt tgctaaacag ttgatattga ggtatcattt tatcgtaaaa
     4921 gacatttttg ctcaacaatt gcttgacgga aatcaacaaa ttttagcatt ttgtaaaaaa
     4981 gtcgctatat aatttggtga attggagtta ttttcatatt tttgcatccc gaagagtttc
//
