LOCUS       sequence-252172-        5151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide vector for allelic replacement in Bacteroides, erythromycin 
            selection and aTC-inducible ss-Bfe1 counterselection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5151)
  AUTHORS   Garcia-Bayona L, Comstock LE
  TITLE     Streamlined Genetic Manipulation of Diverse Bacteroides and 
            Parabacteroides Isolates from the Human Gut Microbiota.
  JOURNAL   mBio. 2019 Aug 13;10(4). pii: mBio.01762-19. doi: 
            10.1128/mBio.01762-19.
  PUBMED    31409684
REFERENCE   2  (bases 1 to 5151)
  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: "2019-08-13"; volume: "10(4). pii: mBio.01762-19. doi"; pages:
            " 10.1128/mBio.01762-19"
COMMENT     Sequence Label: pLGB13
FEATURES             Location/Qualifiers
     source          1..5151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    63..81
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(2572..2640,2641..3432)
                     /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(2790..2809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            3563..3672
                     /note="incP origin of transfer"
     rep_origin      3843..4231
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(4258..4881)
                     /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     4789..4808
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
ORIGIN
        1 tctctatcac tgatagggat aaagtttgga agataaagct aaaagttctt atctttgcag
       61 tctccctatc agtgatagag acgaaataaa gacatataaa agaaaagaca catgaagaaa
      121 attttatctt tgcttgtgat ggctattgta gctatacagt tctcatttgc cggcgcttat
      181 agaaatgaat ataaaacgca aagtgaattt aatgtcatct tggaacgggg ggatgactat
      241 gaaggatttg tagttggctt gggatatact tggatgagta gcaaggtaat tctaccggta
      301 aatcaaaacg gatggagtcc aatatctcgg aatgtttcgg tggacgagag ttttcatacg
      361 atagtctcag aaaggaaata cgatacctcc caatatgcct atgaaaaaag tctgatgcaa
      421 gatccaacaa aagtttccga aaaggtccgt gacttaatag ttaaaaacaa aggtaataat
      481 atcactgaga taaatttagg ccaagaaaag caatatttgc ccacggataa tagtcagata
      541 agtattgtaa ttactgacac tggcagtcgg tacgaaattg tgattagtgc cacggataac
      601 tcaaatggaa agacttatga ggcaaagtat gagagtttga cggacttggt gtcagcggta
      661 cgcgattctg gtagcctgcc tgctgtaaat aaggaaggac ccaatctgga aggactggca
      721 gggttaggat tcggaattgc tgaaacagcc ggaaattggg ctgagaagat tatggataat
      781 cgaggtgcgt acttacctaa gcagatgcgt ttctcgccca aaacgcttcc gccgattata
      841 agattgcctt tagggaacta tcaagtcccc gctaaaggta tgagtagagt gcgtggagta
      901 ggtaaagctt tgggatgggc aggaatggtg cttactggct atcaagttgt gcgtgatgtg
      961 caaaatggtc gatttgccgt ggcaggtaca agaattgccg tagcaggttt agcttatggc
     1021 gttactttta ttccctatgt cggttgggtc ttagctatcg gtatcggagt ggctgattat
     1081 acttggggtg acgagttcta tgactggata gacaatagag cttctgaatt ggaaatgtgg
     1141 tgggacggtg taagattagc attatgagtg gatcccccgg gctgcaggaa ttcgatatca
     1201 agcttatcga taccgtcgac taattgccta tcttccagtg atggaacagc atttgtgcat
     1261 tggctgcaac aatcagcctt acttgtgcct gttctatttc cgaaccgacc gcttgtatga
     1321 atccaacagt tgcatgtggc ctattgtttt tagccgttaa atattttata actattaaat
     1381 agcgatacaa attgttcgaa actaatattg tttatatcat atattctcgc atgttttaaa
     1441 gctttattaa attgattttt tgtaaacagt ttttcgtact ctttgttaac ccatttcatt
     1501 acaaaagttt catatttttt tctctcttta aatgccattt ttgctggctt tctttttaat
     1561 acaattaatg tgctatccac tttaggtttt ggatggaaat aatacctagg aatttttgct
     1621 aatatagaaa tatctacctc tgccattaac agcaatgcta gtgatctgtt tgtatctaat
     1681 aacattttag caaaaccata ttccactatt aaataactta ttgtggctga actttcaaaa
     1741 acaatttttc gaattatatt tgtgcttatg ttgtaaggta tgctgccaaa tattttatat
     1801 ggattgtggc taggaaatgt aaatttcagt atatcatcat ttactatttg atagttagga
     1861 taatttaaga gcttattacg agttacctca cataatttag aatcaatttc tatcgccgtt
     1921 acaaaattac atctctttac caatccagca gtaaaatgac ctttccctgc acctatttca
     1981 aagatgttat ctttttcatc taaacttatg caattcatta ttttttctat gtgatatttt
     2041 gaagtaataa aattttgact atcttttata tttactttgt tcattataac ctctccttaa
     2101 tttattgcat ctcttttcga atatttatgt tttttgagaa aagaacgtac tcatggttca
     2161 tcccgatatg cgtatcggtc tgtatatcag caactttcta tgtgtttcaa ctacaatagt
     2221 catctattct catctttctg agtccacccc ctgcaaagcc cctctttacg acataaaaat
     2281 tcggtcggaa aaggtatgca aaagatgttt ctctctttaa gagaaactct tcgggatgca
     2341 aaaatatgaa aataactcca attcaccaaa ttatatagcg acttttttac aaaatgctaa
     2401 aatttgttga tttccgtcaa gcaattgttg agcaaaaatg tcttttacga taaaatgata
     2461 cctcaatatc aactgtttag caaaacgata tttctcttaa agagagaaac acctttttgt
     2521 tcaccaatcc ccgactttta atcccgcggc catgattgaa aaaggaagag tatgagtatt
     2581 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2641 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2701 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2761 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2821 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2881 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2941 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3001 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3061 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3421 aagcattggt aactgtcaga ccaagtttac tcataacgcg tcaattcgag ggggatcaat
     3481 tccgtgatag gtgggctgcc cttcctggtt ggcttggttt catcagccat ccgcttgccc
     3541 tcatctgtta cgccggcggt agccggccag cctcgcagag caggattccc gttgagcacc
     3601 gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca
     3661 cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg
     3721 caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc
     3781 ccgaagcagg gttatgcagc ggaaaacgga attgatccgg ccacgatgcg tccggcgtag
     3841 aggatctgaa gatcagcagt tcaacctgtt gatagtacgt actaagctct catgtttcac
     3901 gtactaagct ctcatgtttc acgtactaag ctctcatgtt taacgaacta aaccctcatg
     3961 gctaacgtac taagctctca tggctaacgt actaagctct catgtttcac gtactaagct
     4021 ctcatgtttg aacaataaaa ttaatataaa tcagcaactt aaatagcctc taaggtttta
     4081 agttttataa gaaaaaaaag aatatataag gcttttaaag cttttaaggt ttaacggttg
     4141 tggacaacaa gccagggatg taacgcactg agaagccctt agagcctctc aaagcaattt
     4201 tgagtgacac aggaacactt aacggctgac atgggaattc ccctccaccg cggtggctta
     4261 agacccactt tcacatttaa gttgtttttc taatccgcat atgatcaatt caaggccgaa
     4321 taagaaggct ggctctgcac cttggtgatc aaataattcg atagcttgtc gtaataatgg
     4381 cggcatacta tcagtagtag gtgtttccct ttcttcttta gcgacttgat gctcttgatc
     4441 ttccaatacg caacctaaag taaaatgccc cacagcgctg agtgcatata atgcattctc
     4501 tagtgaaaaa ccttgttggc ataaaaaggc taattgattt tcgagagttt catactgttt
     4561 ttctgtaggc cgtgtaccta aatgtacttt tgctccatcg cgatgactta gtaaagcaca
     4621 tctaaaactt ttagcgttat tacgtaaaaa atcttgccag ctttcccctt ctaaagggca
     4681 aaagtgagta tggtgcctat ctaacatctc aatggctaag gcgtcgagca aagcccgctt
     4741 attttttaca tgccaataca atgtaggctg ctctacacct agcttctggg cgagtttacg
     4801 ggttgttaaa ccttcgattc cgacctcatt aagcagctct aatgcgctgt taatcacttt
     4861 acttttatct aatctagaca tattcgttta atatcataaa taatttattt tattttaaaa
     4921 tgcgcgggtg caaaggtaag aggttttatt ttaactacca aatgttttcg gaagtttttt
     4981 cgcttttctt tttctatcgt ttctcagact ctcttagcga aagggaaaga aggtaaagaa
     5041 gaaaaacaaa acgccttttc tttttttgca cccgctttcc aagagaagaa agccttgtta
     5101 aattgactta gtgtaaaagc gcagtactgc ttgaccataa gaacaaaaaa a
//
