LOCUS       sequence-242372-        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"
     oriT            102..211
                     /note="incP origin of transfer"
     rep_origin      382..770
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(797..1420)
                     /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     1328..1347
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    1691..1709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1753..1771
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(4262..4330,4331..5122)
                     /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(4480..4499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caattcgagg gggatcaatt ccgtgatagg tgggctgccc ttcctggttg gcttggtttc
       61 atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc ctcgcagagc
      121 aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct
      181 cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa
      241 gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga
      301 aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaacggaa ttgatccggc
      361 cacgatgcgt ccggcgtaga ggatctgaag atcagcagtt caacctgttg atagtacgta
      421 ctaagctctc atgtttcacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
      481 aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc
      541 atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta
      601 aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc
      661 ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta
      721 gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca tgggaattcc
      781 cctccaccgc ggtggcttaa gacccacttt cacatttaag ttgtttttct aatccgcata
      841 tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga
      901 tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag
      961 cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga
     1021 gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt
     1081 cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc
     1141 gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc
     1201 tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg
     1261 cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta
     1321 gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta
     1381 atgcgctgtt aatcacttta cttttatcta atctagacat attcgtttaa tatcataaat
     1441 aatttatttt attttaaaat gcgcgggtgc aaaggtaaga ggttttattt taactaccaa
     1501 atgttttcgg aagttttttc gcttttcttt ttctatcgtt tctcagactc tcttagcgaa
     1561 agggaaagaa ggtaaagaag aaaaacaaaa cgccttttct ttttttgcac ccgctttcca
     1621 agagaagaaa gccttgttaa attgacttag tgtaaaagcg cagtactgct tgaccataag
     1681 aacaaaaaaa tctctatcac tgatagggat aaagtttgga agataaagct aaaagttctt
     1741 atctttgcag tctccctatc agtgatagag acgaaataaa gacatataaa agaaaagaca
     1801 catgaagaaa attttatctt tgcttgtgat ggctattgta gctatacagt tctcatttgc
     1861 cggcgcttat agaaatgaat ataaaacgca aagtgaattt aatgtcatct tggaacgggg
     1921 ggatgactat gaaggatttg tagttggctt gggatatact tggatgagta gcaaggtaat
     1981 tctaccggta aatcaaaacg gatggagtcc aatatctcgg aatgtttcgg tggacgagag
     2041 ttttcatacg atagtctcag aaaggaaata cgatacctcc caatatgcct atgaaaaaag
     2101 tctgatgcaa gatccaacaa aagtttccga aaaggtccgt gacttaatag ttaaaaacaa
     2161 aggtaataat atcactgaga taaatttagg ccaagaaaag caatatttgc ccacggataa
     2221 tagtcagata agtattgtaa ttactgacac tggcagtcgg tacgaaattg tgattagtgc
     2281 cacggataac tcaaatggaa agacttatga ggcaaagtat gagagtttga cggacttggt
     2341 gtcagcggta cgcgattctg gtagcctgcc tgctgtaaat aaggaaggac ccaatctgga
     2401 aggactggca gggttaggat tcggaattgc tgaaacagcc ggaaattggg ctgagaagat
     2461 tatggataat cgaggtgcgt acttacctaa gcagatgcgt ttctcgccca aaacgcttcc
     2521 gccgattata agattgcctt tagggaacta tcaagtcccc gctaaaggta tgagtagagt
     2581 gcgtggagta ggtaaagctt tgggatgggc aggaatggtg cttactggct atcaagttgt
     2641 gcgtgatgtg caaaatggtc gatttgccgt ggcaggtaca agaattgccg tagcaggttt
     2701 agcttatggc gttactttta ttccctatgt cggttgggtc ttagctatcg gtatcggagt
     2761 ggctgattat acttggggtg acgagttcta tgactggata gacaatagag cttctgaatt
     2821 ggaaatgtgg tgggacggtg taagattagc attatgagtg gatcccccgg gctgcaggaa
     2881 ttcgatatca agcttatcga taccgtcgac taattgccta tcttccagtg atggaacagc
     2941 atttgtgcat tggctgcaac aatcagcctt acttgtgcct gttctatttc cgaaccgacc
     3001 gcttgtatga atccaacagt tgcatgtggc ctattgtttt tagccgttaa atattttata
     3061 actattaaat agcgatacaa attgttcgaa actaatattg tttatatcat atattctcgc
     3121 atgttttaaa gctttattaa attgattttt tgtaaacagt ttttcgtact ctttgttaac
     3181 ccatttcatt acaaaagttt catatttttt tctctcttta aatgccattt ttgctggctt
     3241 tctttttaat acaattaatg tgctatccac tttaggtttt ggatggaaat aatacctagg
     3301 aatttttgct aatatagaaa tatctacctc tgccattaac agcaatgcta gtgatctgtt
     3361 tgtatctaat aacattttag caaaaccata ttccactatt aaataactta ttgtggctga
     3421 actttcaaaa acaatttttc gaattatatt tgtgcttatg ttgtaaggta tgctgccaaa
     3481 tattttatat ggattgtggc taggaaatgt aaatttcagt atatcatcat ttactatttg
     3541 atagttagga taatttaaga gcttattacg agttacctca cataatttag aatcaatttc
     3601 tatcgccgtt acaaaattac atctctttac caatccagca gtaaaatgac ctttccctgc
     3661 acctatttca aagatgttat ctttttcatc taaacttatg caattcatta ttttttctat
     3721 gtgatatttt gaagtaataa aattttgact atcttttata tttactttgt tcattataac
     3781 ctctccttaa tttattgcat ctcttttcga atatttatgt tttttgagaa aagaacgtac
     3841 tcatggttca tcccgatatg cgtatcggtc tgtatatcag caactttcta tgtgtttcaa
     3901 ctacaatagt catctattct catctttctg agtccacccc ctgcaaagcc cctctttacg
     3961 acataaaaat tcggtcggaa aaggtatgca aaagatgttt ctctctttaa gagaaactct
     4021 tcgggatgca aaaatatgaa aataactcca attcaccaaa ttatatagcg acttttttac
     4081 aaaatgctaa aatttgttga tttccgtcaa gcaattgttg agcaaaaatg tcttttacga
     4141 taaaatgata cctcaatatc aactgtttag caaaacgata tttctcttaa agagagaaac
     4201 acctttttgt tcaccaatcc ccgactttta atcccgcggc catgattgaa aaaggaagag
     4261 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4321 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4381 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4441 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4501 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4561 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4621 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4681 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4741 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4801 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4861 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4921 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4981 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5041 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5101 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcataacgcg t
//
