LOCUS       sequence-273919-       10978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pNBU2-derived shuttle vector for integration at attBT2 sites in 
            Bacteroides chromosomes, inulin selection for use in anbitiotic 
            resistant strains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10978)
  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 10978)
  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: pLGB28
FEATURES             Location/Qualifiers
     source          1..10978
                     /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"
     oriT            560..669
                     /note="incP origin of transfer"
     CDS             complement(join(800..1591,1592..1660))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAVPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1423..1442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(10930..10946)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10930..10946)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc
       61 tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag
      121 ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg
      181 atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag
      241 tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga
      301 gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat
      361 caacaggttg aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatcaa
      421 ttccgttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
      481 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
      541 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
      601 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
      661 gctggccggc taccgccggc gtaacagatg agggcaagcg gatggctgat gaaaccaagc
      721 caaccaggaa gggcagccca cctatcacgg aattgatccc cctcgaattg acgcgttatg
      781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
      841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtactgcaa
     1021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatcat
     1681 ggccgcggga ttaaaagtcg gggattggtg aacaaaaagg tgtttctctc tttaagagaa
     1741 atatcgtttt gctaaacagt tgatattgag gtatcatttt atcgtaaaag acatttttgc
     1801 tcaacaattg cttgacggaa atcaacaaat tttagcattt tgtaaaaaag tcgctatata
     1861 atttggtgaa ttggagttat tttcatattt ttgcatcccg aagagtttct cttaaagagt
     1921 gatctggaag aagcaatgaa agctgctgtt aagtctccga atcaggtatt gttcctgaca
     1981 ggtgtattcc catccggtaa acgcggatac tttgcagttg atctgactca ggaataaatt
     2041 ataaattaag gtaagaagat tgtaggataa gctaatgaaa tagaaaaagg atgccgtcac
     2101 acaacttgtc ggcattcttt tttgttttat tagttgaaaa tatagtgaaa aagttgccta
     2161 aatatgtatg ttaacaaatt atttgtcgta actttgcact ccaaatctgt ttttaacata
     2221 tggcactaat gcttagtatt atgaagaaca aaaaacttct ttgtagcgtt tgtttcctgt
     2281 ttactttcat gtcagtcctt tgggggcaaa acataacagt gaaaggaaac gttacttcaa
     2341 agactgatgg tcaaccaatt attggtgctt ccgtagtgca aaatgattct aaatcaactg
     2401 gaactatcac agatttggat ggtaacttta ctcttagtgt accaacgaat gcattattgg
     2461 caatttctta tataggatat aaagaagtaa ttattgccgc taaaccttca ttgaagatag
     2521 tgatggaaga ggatgctaag atgattgacg aagtggttgt gacaggttat atggctgaaa
     2581 agaaagccag tctgacaggt tcggtagctg tagtgaagat gaaagaggtg gctgacatac
     2641 ctactggtaa tgtgatgtca ggtttgcagg gacgtgtagc cggtatgaac gtaacaactg
     2701 atggtaaacc cggaggtggt aacacagata ctaagttacg tggtattact accatcaaca
     2761 attcttctcc tctttatgta atagacggtg ttcagacaca tgataatgtg gcttctatta
     2821 tctcatccaa tgatgttgaa tccattcagg tgttgaaaga tgcagcttcg gctgccattt
     2881 atggtgcgca agctgccaat ggtgtcatta ttatcaccac aaaacgtgct aaggaaggcg
     2941 atgtaaaagt aagttttgat atgtcgctca ctgcccagac ttttactggt ggtattgata
     3001 tgcttgacgc ttaccaatgg ggtgatgtgt attggcaggc ttataaaaat acttttggaa
     3061 ccaatcccaa tagtgtagta tatggaaatg gagaaaaggc ccaactgcaa gaatattatt
     3121 ttgaccagaa tggagtgaaa ataaagtcgg gcaacactga ttgggcaaag gaaatatttg
     3181 gtacggcttt aatgcaaaat tataatcttt ctctttctaa agggtttaag gatgggaatg
     3241 tggctcttac cttaaattat atgaatcagg atggtctttg ccgtaataca gactacgaac
     3301 gttttaattc gcgtttagcc tctaattttc gctttgtaga tggtaaagtg cgtgtgggtg
     3361 aaagtatttc ggtgaatcac tggttggagc atcttaatcc tgccggtata gaagaattgg
     3421 ttattgccca gcatccggcc attcctgtgt atgatgaaaa tggagggtat gccggtggct
     3481 atgtggatat attgggggat aagccgaatg ccattcgctt gacagacaat gaagccaata
     3541 atcgtcacaa aagttggcgt atctttggca atgtttattt ggaagtggaa cccattaaaa
     3601 atctagtgtt gaagagtcaa tttggtttga actatactac cggtttcaac tcaactttcg
     3661 tccccaaatg gagtgaagcc gatcgtaagg tagatgtaaa tgaactgacc gtacgacaag
     3721 ataatagcgt gcaatgggta tggtcgaaca ctgccaatta tagtatagac tttgggaaga
     3781 acaatatcag tgcagtgata ggaactgaag ctaagaaaga gaatggtgaa catttacagg
     3841 gatatggtcg cggattggtg atagaagatc tcgattatcg ttacttagac actgtaactg
     3901 aaggaaaaag tgtgagtaac aatgcttcta cctatgctat ggtctcttac tttggtaagg
     3961 tgaactatgc gtttgatgat aaatacctta tatcaggaac agtgcgtcgt gatgcttctt
     4021 cacgtttcgg taaagggaat aattctgcta tctttccttc agtatctgcc ggttggagag
     4081 tatcgcgtga aaaatttatg gaatgtacgc aaaactggct ttcagatctt aaattgcgtg
     4141 catcgtgggg gatcaatggt aatgatcaga ttgataacaa tgctacttac aacctctatt
     4201 acactaacat gtccaatggc tcatacaatt tcaatgggga tggtacaacg gtattgcccg
     4261 gagtacagaa agaccgttcg ggtaacaacg acctgaaatg ggaagagacc aaacaactga
     4321 actttggtat tgatgctgct ttctttgaca accgtttggg actgacactg gattattttc
     4381 aaaaaaagac tactggtatg ttaatccaga aaccttatat aggagtgatc ggtgaaggtg
     4441 gatataaatg gtacaatgct gccagtatgg ataatagcgg tgtggaagca acctttactt
     4501 ggcgagatga aataaaggat ttcaagtatg atatttcgtt caacctttcc tattataaaa
     4561 atgaaattac cgaacttccg gatgtaatca aatatacttg gggaggtggt aatggtgtgg
     4621 acaaaactat tgtaggtcaa ccttacggat catggatgtc ctataaagct gatggtgtat
     4681 tcaaaacaaa acaggaggta tatgaatacc taagcaaata tgatgtacag attggtgctc
     4741 ccggcgtagg acgtattcgc tacaaagacc ttaacggtga taacattatc aatactgctg
     4801 atatggattg gcaagggagt gaccaaccta gatttattgg tggattgaac atcggtgcat
     4861 cttacaaagg atttgacctt tctgtattct tcaatggtat gattcgcgat gcatggaaca
     4921 attccaagtt ttataccgac ctttttcaag gttggacagg taatcactct gtacgtttga
     4981 tggatgcaat gcatgcttgg aatgcttatg agcaaacggg agtttataat tgtgatatcc
     5041 ctgcattgac tgtagttgat aataatgtag aaacacgcag ttccactttc tttattgaag
     5101 atggttctta tgtgaagttg aaaactctaa ctgtgggtta tactttcccg gataagtgga
     5161 ttaagaaagc gcatctttcc aatttgcgtt tgtatctgca agcacagaac gtttttacgc
     5221 tgactaatta tacgggtgcc gacccggaag gattgggcta tccatatccc caacctcgta
     5281 cctatacatt cggtctatca cttggttttt aatttttatc ctaaatacag aacattatga
     5341 aattgaaata tatctttgca gtaggtgtga tggcactatt ggcaagttgc gatgagagtc
     5401 gttttctaga tattaaacct cagggtacac tcaatgacga tttgttatca tcggccgatg
     5461 gtgtagagct tttggttact tcagcttatg caggattgaa aggtcctaat cgtgatatgc
     5521 actgggtgcc gatgaccaat tggacgtatg gtgaagtacg ttcggataat gcctacaaag
     5581 gtggtggcgg tgtaaatgat ggagactttt gtacgctgct tgaaaccttt aaaatagatg
     5641 ctacatgggg caatgccgac gaaaaatggt ttcaacttta ttgttgcttg caacgttgta
     5701 acagcgcatt gcgggtaatg aatacgcttg atgagaacgc tttgccgatg attaaaagtc
     5761 gtaaagccga aatgaaagtg attcgtgcac acttcttctt tgaactggtt cgccttttca
     5821 agcaagtacc ttattttgat gagaatgtgg aaaatgatga ctataagtac attcccaaca
     5881 atcagtttac ccgtgaagaa cttctgggta aaatagcaca agaatttctt gatgctgcta
     5941 acgatttgcc tgatactcaa tctcaagtag ggcgtgtcac taagaacatt gcttatgctt
     6001 atgctgccaa agctagactt tatcaggctt accgacaaga tgataataac caagtggttg
     6061 ctattgataa acaattgtta gctgaagtgg tgactctttg cgacaaagta ggtgcgcaag
     6121 gaaaggcata tactcttttg aacgattttc agaaacttga tcaattggaa tatgaaaatg
     6181 gggtagagtc tgtgtttgct gtacaatatt cgatggatga tgaaaccgat ggtgccggga
     6241 atataaactg gagtaatctg ttgaatgctc ctaaaggtcc ttatggtggc gacggatttt
     6301 tcttacccag tcagaacttg atcaatgcct ataagacaga taactatgga ttgcctttgt
     6361 ttgatacttt caacgccgat gactatggaa tatacgaagg aagtgagctc accaacattg
     6421 atgcaactgt ggatcctcgt ttggatttcg ttaccggtcg tcctggtatc acttggaaaa
     6481 catataccgt tgagccttgt aaagcaaact ggatacgtaa cagcggtgaa tatggcttta
     6541 attgtacaaa gcgcttctac atctcgcctg aaagtgaaga tatgttccaa ggttggcctt
     6601 ggggagcttc tcatctcaac tggcagataa ttcgctatgc tgatgtgttg ttatggaagg
     6661 ctgaagcctt gatagagatt ggcgaagcta ccggactgga agaggctcgc aaaattatta
     6721 accgtattcg tctgcgtgcc aaaaatagtc cttatgtaaa agactttgaa cattccgaac
     6781 aggatgcagc taattatctg ataggcgaat atcctgctga gggctggagt caggactttg
     6841 cccgcaaggc attacgttgg gaacgccgta tggagtttgc tatggaaggt gaccgtttct
     6901 tcgacttggt acgttggggg attgctgcgg aaaccatgaa cagctacatt agtgtggagc
     6961 agaataagcg cgtttattat cgtggtgctc gctttgttcg tggcagagat gaatatttgc
     7021 ctattcccaa tgcacaatat aacttctctg aaggcaatta tgtgcagaat cccggatacg
     7081 gtaagtttaa ctagctctaa taccatctga catgaatgta ttattctcgt ttaaacaagt
     7141 acagaccttt ttgactatcc tggttatgat gattttctca tttttggatg cttcggccga
     7201 tacttcttcc ttattaataa aagatttggg tgaaggacat tgcctggtac agataaatac
     7261 aaatcaacgt tatttgcttt tacctgtaga ggaggtaatg cctgatgtac gtgtcagcat
     7321 gatcgttaat aataaagagg tgaatgttgc ggatgttcgt ctggcagtga acagagtgga
     7381 ctattttgtt ccgctggatc tttcggctta tgctggtaaa aacatactgt tgaaatttaa
     7441 attaggctcg aatgatccta tacgggggaa actttctgct gtttgttgca aggaaatgaa
     7501 attgtccgac acgttcgata cgggcaaccg tgaaaagttc cgtcctactt atcatttctc
     7561 acctctttat gggtggatga atgatcctaa tggaatggta tataaagacg gtgaatacca
     7621 ccttttctat caatacaatc cttatggttc caaatgggga aatatgagtt ggggacatgc
     7681 tatcagccaa gatcttgtaa actggaaaca tcttcctgta gctattgctc ccgatgcatt
     7741 ggggacaatt ttcagtggct cggcagttgt cgattttgat aatacagccg gttttggtgc
     7801 aggtgccatt atcgccatct atacacaaaa cagtgaccga caagtacaaa gcattgctta
     7861 cagtactgac aacggacgta ctttcaccaa atatgagaat aatccggtac tgacgtccga
     7921 agcgcgtgac ttccgcgacc cgaaagtatt ttggtatgaa aacaccaaac gctggatcat
     7981 ggtattagca gtagggcagg agatgcagat attctcttct cctaatctga aagattggac
     8041 ctttgaaagt cgtttcggtg aagggcaagg tgcacatggt ggtgtttggg aatgtcccga
     8101 tttgtttgaa cttccagtag aaggtaccaa tgataagaaa tgggtgctgc tttgtaattt
     8161 gaatcccgga ggtcctttcg gaggaagtgc tactcaatat tttgtaggct ccttcaacgg
     8221 aaaagagttt gtaaacgaat tcccttccaa aaccaaatgg atggactggg gaaaagacca
     8281 ttacgccacc gtaacttgga gcgatgctcc cgacaaccgc cgtatcgcta tcgcatggat
     8341 gagcaactgg caatacgcaa acgatgttcc tacctcccaa taccgtagcc cgaattccgt
     8401 tccccgtgac ttgagcctgt ttaccgtaga tggtgagact tacctccaat cagctccctc
     8461 tcccgaactg ctggctttgc gtgatgcttc caagaaacgc tcgttcaaag taaacggaac
     8521 acgcacaatc aaagaaatga ttcccggcaa tgacggtgca tacgaaatag aattgacaat
     8581 cgaaaatcaa cgtgcagacg taatcggctt ccgtctctac aatgacaaag gcgaagaagt
     8641 ggatatgcag tatgacatga aagaaaagaa attctcaatg gatcgtcgca agagcggaga
     8701 ggtcagcttc aatgagaatt tcccgatgtt gacctggaca gcaattgagc aaggtggaaa
     8761 cgaggaacaa ggcggcaaag aggcattgaa actccgtctg tttgtggata aatccagtgt
     8821 ggaagccttt ggaaatggtg gacgttttgt gatgaccaat caggtattcc cttcggaacc
     8881 ttacaatcat atcaactttt acagtaaagg aggagcttat aaagtagatt cgtttgtggt
     8941 tcataagttg aagccataaa caataggcca catgcaactg taaatgttta cgcgtcctcg
     9001 gtaccaacaa atactttcag gacgatgtaa agtcctgaga aatcaaactg aatggataga
     9061 taaatattag tcttttctca tcggattctt ttaatatcgc tactctttta ttatgtagtg
     9121 gttgtattat cttaaataaa acaaaaacac ccgactttac attcttgggt gtaaaacagg
     9181 gtgtttgttt tgcaatctgc tgattttcag cgttttttgc ggagagacag gctctcgaac
     9241 ttatactttc acagaatatc ataaaactgc aaacgactga taatcaagtg taaattacaa
     9301 tatagagtaa aagtaaatgt ttctaaatgt cttttgctat ctcaataatt gggtgtatat
     9361 ttgggtgtag aatttcaaac gcacccaatt atgaatatca agcgcaacat catttttgca
     9421 ttggagagcc ggaaaaagaa cggtgtgcca atcgtagaga acgtacccat ccgtatgcgt
     9481 gtcatctttg ccagccaacg tatcgagttt acaacgggct accggattga cgtagccaaa
     9541 tgggatgcag ataagcagcg ggtaaagaac ggatgtacca acaagctaaa gcaaagtgca
     9601 gccgaaatca atacggactt gctgaaatac tatgccgaaa tccagaatat tttcaaggaa
     9661 tttgaggtgc aggaggtcat gccaacgacc caacagttga aggaagcttt caacatgaga
     9721 atgaaagaca ccagtgaaga acagccggaa gaagcccctg tcagcttttg ggaggtgttc
     9781 gatgagtttg taaaagagtg cggtaaccag aataactgga cggcatccac ctatgaaaaa
     9841 tttgcagcag tgaggaacca cctcaaagag ttcaaggagg atgcaacgtt caactatttc
     9901 aacgagtttg gattgaacga atacgtcaac ttcctgcgtg acaccaagga tatgagaaac
     9961 agcaccatcg gcaagcaaat gggattcctc aaatggttcc tgcgctggag cttcaagaaa
    10021 ggacatcatc agaacattgc atacgatacg ttcaaaccga aactgaaaac cacctcgaaa
    10081 aaagtaatct tcctgacttg ggatgaactg aacaagctga aagactacca gatacccaag
    10141 gataagcaat acctggaacg tgtgcgtgat gttttcctgt tctgctgctt tacgagtttg
    10201 cggtattcgg atgttcgcaa tctgaaaaga agcgatgtga agtccgacca catcgaaata
    10261 accacagtca agactgccga cagcctgacg attgaactga acaaatacag caaagccata
    10321 ctggacaaat acaaggacat ccatttcgag aattacatgg ctctgcccgt catcagcaac
    10381 cagaagatga acgattacct gaaagagctg ggcgaactgg cagaaatcaa cgagcctgta
    10441 cgggaaacct actacaaggg aaatgaacgt attgatgaag tcacacccaa atacgctttg
    10501 ctcagtaccc atgcaggaaa aaggacattc atctgcaatg cgctggctct cggaatcccg
    10561 gcacaggtgg tcatgaaatg gacgggacac agcgactaca aagctatgaa accctacatt
    10621 gacatagcgg atgatattaa ggcaaatgcc atgaacaagt ttaatcaact ataaaagtat
    10681 caaacagttt catagatagt taccaatgac tatatttgta gaaattatca tcacaatatg
    10741 agcaacaagg aatccaacgt agagaaaacg aattttgatg cattcataca agcggtcggt
    10801 tcggaaatag aacaggcaca agtaaggctg attgttgcag ccaatgcaca aatgctgttc
    10861 catcactgga agataggcaa ttagtcgacg gtatcgataa gcttgatatc gaattcctgc
    10921 agcccggggg atccactagt tctagagcgg ccgccaccgc ggtggagggg aattccca
//
