LOCUS       sequence-242370-        6609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide vector for allelic replacement in Bacteroides, tetracycline 
            selection and rhamnose-inducible ss-Bfe1 counterselection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6609)
  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 6609)
  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: pLGB30
FEATURES             Location/Qualifiers
     source          1..6609
                     /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             join(5720..5788,5789..6580)
                     /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(5938..5957)
                     /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 tcatgtttaa 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 cctccacctt attcttcccg accgaacgag cggtattcaa gcggagtaag cccggtacgt
      841 tttttgaaaa aggagaaaaa atattcggta gactggaagc ccagaaaaaa agaaatttct
      901 tttactgact gggaagttcc caccaacatt tgcttggctt tacgaagttt cagttcctga
      961 aaatatttgg cgggtgcata gccggtatat tccttaaaaa ccctgcggaa ccatgagtaa
     1021 ctgatattca gtctcatcgc cagttcctcc ggatcaatat tgccggatac attttcattc
     1081 ataagtatct ttgcctgctc gatcttctga tcgacatcac tcatctcgaa gaccttattc
     1141 ttggaaatcg aaagaatcat accaatcatg tgaagtacga ttccggaaag gtattgctgg
     1201 gcagatattt tatccgcttc tgccacttcc aaagcgcggg aaaacaaaga taccagttct
     1261 tcattaatac cgacttccaa tatctgtctt tcctgagaaa gaaaggcatc gcccacaatc
     1321 gtatcaatcg ccggaccttc aaaaccaatg taatattccg tccatccggt ctgccgcaat
     1381 ggataataag tatgccactg tccgggaaac aacaccatca gccgaccttt acaaacttgt
     1441 ctttcaggag tagaatcgga agaaaacaaa ccgcgccctt ttgtaatata gaccaattga
     1501 tattcccgca atactcttcc tttttcggca ttgaaatagt aaccggacgg atgctccttc
     1561 agcggataag gagaatcggg agggatagac tgatagccaa ccgtattcac ccacagaccg
     1621 aatttgcggt ccatgtcatt aactatcaag tatttaaatt ctaatcccaa attattatcc
     1681 tcagtcatac aggcgtgttt tcatacaatt gaattgaaca aaagtaggaa ttattttcaa
     1741 aaaacacacg ctaaacagta aaaaaacatc cagagcacct gtccagatcc tccggatgct
     1801 tattatttcg gagaaaacat gcataaatca tgcttttttt gcataaaaag taaaatttat
     1861 actgatgtaa ggtttggcta tgcagatttg tgtcaaaatg cacatccttt ctatcaaaat
     1921 gcgtaaggaa aaggaggaag gaaccgccta tctttgcaat gtaggtaaat ggatacctta
     1981 aatatataga caaaatacca tgaagaaaat tttatctttg cttgtgatgg ctattgtagc
     2041 tatacagttc tcatttgccg gcgcttatag aaatgaatat aaaacgcaaa gtgaatttaa
     2101 tgtcatcttg gaacgggggg atgactatga aggatttgta gttggcttgg gatatacttg
     2161 gatgagtagc aaggtaattc taccggtaaa tcaaaacgga tggagtccaa tatctcggaa
     2221 tgtttcggtg gacgagagtt ttcatacgat agtctcagaa aggaaatacg atacctccca
     2281 atatgcctat gaaaaaagtc tgatgcaaga tccaacaaaa gtttccgaaa aggtccgtga
     2341 cttaatagtt aaaaacaaag gtaataatat cactgagata aatttaggcc aagaaaagca
     2401 atatttgccc acggataata gtcagataag tattgtaatt actgacactg gcagtcggta
     2461 cgaaattgtg attagtgcca cggataactc aaatggaaag acttatgagg caaagtatga
     2521 gagtttgacg gacttggtgt cagcggtacg cgattctggt agcctgcctg ctgtaaataa
     2581 ggaaggaccc aatctggaag gactggcagg gttaggattc ggaattgctg aaacagccgg
     2641 aaattgggct gagaagatta tggataatcg aggtgcgtac ttacctaagc agatgcgttt
     2701 ctcgcccaaa acgcttccgc cgattataag attgccttta gggaactatc aagtccccgc
     2761 taaaggtatg agtagagtgc gtggagtagg taaagctttg ggatgggcag gaatggtgct
     2821 tactggctat caagttgtgc gtgatgtgca aaatggtcga tttgccgtgg caggtacaag
     2881 aattgccgta gcaggtttag cttatggcgt tacttttatt ccctatgtcg gttgggtctt
     2941 agctatcggt atcggagtgg ctgattatac ttggggtgac gagttctatg actggataga
     3001 caatagagct tctgaattgg aaatgtggtg ggacggtgta agattagcat tatgagtgga
     3061 tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacta attgcctatc
     3121 ttccagtgat ggaacagcat ttgtgcattg gctgcaacaa tcagccttac ttgtgcctgt
     3181 tctatttccg aaccgaccgc ttgtatgaat ccaacagttg catgtggcct attgtttgga
     3241 gataagtata tatcccaact gtattgcctt atagaaattt cctgaccgct ccattatttt
     3301 gatgacattg atttttggaa catgaataaa agtttatctt tttcgttcat gcggatatta
     3361 tcagaataac cgccttttgt tatttgatac ccgcatggct taaccataaa aatgcctaag
     3421 cccttagtgt atgaacttac ttctgatgca tagtctttac ttgtatttaa tggaactttc
     3481 cctttaatat gacaccactc attattgcaa ctgatgtctt caatctcaga catcattttt
     3541 tgcaaatctg taatagcttt ggaacttgcc gcttggggta tctgcaactc aaaatagagc
     3601 atcggttcga gaatgtccac acctgactgt tgcaaggcca gcctgaagac ataaggggtc
     3661 agctgtctga aatcagcagg tgtacttacc gggctataat actcggcttg agtaaaagtt
     3721 actttcagat cagtcacttc ccatccatgt aacccggatt ggcaagacat acgaatccct
     3781 tcaaaaacgg cattttgaaa agaatggttc agataaccat aggagatgtc actttcgatt
     3841 tgcaaccctg tccctaacgg taagggttca agagtcagcc ctattgtggc ccaataaggg
     3901 ttgggcggca cttcgatctg aataatctta ttgacctttt ttacaggtcg ttctttgtat
     3961 atagtcttga tctcatcaaa atggaccttt acggaaaatc gttcttccag caatgtctgt
     4021 atgatttcct tttgggttaa accatataac gagatttcca attcatcact atatgagttt
     4081 atggaaaagg acaaagacgg gtcttcaatc cacaatgtat tcagagcgga tatcaccttg
     4141 cttctctctt cgggcctgtc tggccggacg gaggatttga gagcgggatg ctgatgcgat
     4201 aatccttgaa tcaaacaagg ttcagcacct aaataatttc cgattcgaaa atcatccata
     4261 tcctctacaa tcgcgatatc attggcgccc acttcatcaa catttatctc tctgccctga
     4321 ttgatagttt ttagattttt aatcttgatg aatttttccg aatcgttgat tcttacaacg
     4381 tctcgaagtc tcagacttcc gtcaattatt tttagaaaac ttcttttatg tcctttgggg
     4441 tcatgctcta tcttataaag ataagatgaa agtctgtttg agaccgatgc cggaggaagt
     4501 ataaaagaag tgatggcgtc caacaactca ttgataccga tattgaacat tgctgatcca
     4561 tgtagcaccg gatagacttt ggcttttgcc acaagagcga ttatcgtatt ccaataatca
     4621 gccggtgaaa tttcgctatc cgccaaatat cgttctaata tattgtcgtc atggttgcat
     4681 acaaattctt tgtattcttc ctttatatat gtttgggagc aaaccggata aaccgatcca
     4741 tcgacaacat tttgcataaa caggacatct tgagacagat ttgcttttat atccagatac
     4801 aaacgctcca aattcacacc ggctcggtca atcttattga taaatataat tgtcgggatt
     4861 tgcagcttct gtaaagtatt gaacagcaac tttgtctgcg cttgtatgcc ttcctttgcg
     4921 gataagatga ggactgctcc atcaagcatt ttgaatgtcc gctccacttc cgcaataaaa
     4981 tccatgtgtc ccggagtgtc aatgatattg catttcacac cattccagat aatagatgtc
     5041 gtagaagccc gaacagtaat tcctctacgt ttctctatat ccatagagtc cgttatggtg
     5101 tcaccattat ccacacagcc gcacttttcc gttgctccac tggcaaacag cagattctcg
     5161 gttacggaag tttttcctgc atcaatgtga gcaagaattc ctaaatttat aatattcatt
     5221 tggattaagc aataatatac tacaatagat gcattgtcga aacgcacctt ttaatacctc
     5281 ctcgtagcat atgagaacta caggattcct aactcgtatt aatatgtata ttattactgc
     5341 cgcataacga ttacaaaatt acacaaaaaa atatatctaa caaaagtagg aggatttttt
     5401 aactttttac catagacatt ttattaggga aacgtaggtt caactggtaa gtacgtttct
     5461 ctctttaaga gaaactcttc gggatgcaaa aatatgaaaa taactccaat tcaccaaatt
     5521 atatagcgac ttttttacaa aatgctaaaa tttgttgatt tccgtcaagc aattgttgag
     5581 caaaaatgtc ttttacgata aaatgatacc tcaatatcaa ctgtttagca aaacgatatt
     5641 tctcttaaag agagaaacac ctttttgttc accaatcccc gacttttaat cccgcggcca
     5701 tgattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5761 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5821 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5881 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5941 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6001 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6061 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6121 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6181 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6241 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6301 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6361 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6421 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6481 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6541 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6601 ataacgcgt
//
