LOCUS       sequence-375745-        5987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli - Eggerthella lenta shuttle plasmid with kanamycin 
            resistance in E. lenta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5987)
  AUTHORS   Dong X, Guthrie BGH, Alexander M, Noecker C, Ramirez L, Glasser NR, 
            Turnbaugh PJ, Balskus EP
  TITLE     Genetic manipulation of the human gut bacterium Eggerthella lenta 
            reveals a widespread family of transcriptional regulators.
  JOURNAL   Nat Commun. 2022 Dec 9;13(1):7624. doi: 10.1038/s41467-022-33576-3.
  PUBMED    36494336
REFERENCE   2  (bases 1 to 5987)
  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: "Nat 
            Commun."; date: "2022-12-9"; volume: "13(1):7624. doi"; pages: " 
            10.1038/s41467-022-33576-3"
COMMENT     Sequence Label: pXD69m2-AphA
FEATURES             Location/Qualifiers
     source          1..5987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3026..3820
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     complement(3898..3916)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3984..4088
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4089..4157,4158..4949)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4307..4326)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5120..5708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5609..5628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5889..5960)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagcaggg gcattcgaga caaggagatt atagcatggc ggcaaacgct ggcgagacga
       61 aggtgaagct ctgggagggc atctgctacc cagagaacat gcgcgatgac tggcaggacg
      121 agatagacga cattcttcaa gtgcctttcg cttatgcggt tcacgacata gaccatgacc
      181 agaagagcaa gcagcgcaag acgcatgcgc acgtcattgt tgtctggggc ggcaacacga
      241 cacgcaaggc aatcatcaac gtcttgaacc gattgagcgc ggacggcaag aagtgctgtt
      301 cgtcagccga gccaatcaac aacatcaggc atgcgtatga ctatctgatt cacgacacgg
      361 cttcctgtcg caagaagggc aaggagctgt atccggtcga agcacgcatc gagggaaaca
      421 acttcgacat tggtcagctc gaacagctct cgactaagga caagcaggac atgctcttcg
      481 agttggtcgg cttcatcatg tgcgagaggt tcgagaccat caacgatttc acggctgcgg
      541 cgctgaggga gtttcccgag cagtaccgcg aaatcatcgt ggggtacaac tcgattctcg
      601 agcgctattg tcgcgggaac tatctgaacg ccgaacgtaa gcgcaagtgc gtggaacagt
      661 caggtgccaa agagtagtcg ggtcaatgca gcaatgcgtg actacggggc ggcgtggtct
      721 gcgctcgcgc agatctaagc cgactcgttc acggattgcg gaattgacgc gacggccaac
      781 cgaatagacc agacttgatt aagggtcgat ggacgctctg ctccatcgac cctttccttg
      841 cccgtgtctc gactggatga gacctgcaga atcgggcggc tcattatttt tcgctaacta
      901 cgtccgaaaa ataatgtgtc cgaaaaataa tgtgacaaat gacaaatgac aaatgaccac
      961 aggggggcca ctacgacggc ccccctaata gtgccgtgtg ccgtgtgccg aaccgcaggt
     1021 agattgaggg gtggttcgga ggggcgaagt ccctccgttc ggcgctcgcg ccgatggccc
     1081 gagggccgtg agcgaagccg cgaacgggca agggctgggg ggtctcgggg ggaacgcccg
     1141 agcgggtgca gggctggccc tgccacgtcc ttagcgtagc taaggtctaa cgtgctagac
     1201 tttagtcgat taagtctgta cggttcggag gtttccatgg ctcacatcgc gaaatacaag
     1261 gctccatcgg tcgggcacat gcttgcccac taccgccgcg accgcagcag cctggagcgc
     1321 gacaacatcg accccgagcg catcaagaac gacttggtgg tcggccacta cacgaacaag
     1381 gacggcaagc tcgttgtggg gcgcgtggag ccgcgcgatg gcgagccgaa ctggggcacg
     1441 gtggagagcc gcatcgagcg cgtgaacgag gcccagaagg ccgcagggaa gcgtgccacg
     1501 cgcaaggatg cggtcgttat ggctgacgtt gttgtgacgc ttcccgacaa tgtgcgcaag
     1561 ggcgatgagg acaggttctt ccgtctcacg tactggtacc tgtcgaagaa gttcggcatc
     1621 gagaacatga tgggcgggta cgtccacaag gacgaggtgt tgaaggatgg cacgcccgcc
     1681 cgcgaccaca tgcacgtgcc gttcacgccc atcctagatg gacggttcaa ctacaagcag
     1741 atgtgccccc gcagcttcta ccggagcatg cacaaggagc ttggggatta cctagagggg
     1801 cgcatgggct accgccctgc aatcgagctt gacgaggaga cgcgggcgca gcgtgtatac
     1861 acggacaagt ccgtggatat cgacaaggta cgctgtgcgg tcgataaggc cgtggttgaa
     1921 cctgcgaagg acgaagcggc gcgaatcgtc gcggcggcta gggaagaagc cgccgcttta
     1981 ctgaatgatg ccgaaacgcg taaggcagag cttgttaccg agattgccga caaggagggt
     2041 gaccttgccg agcttgacag ccagctcgag gacgtgaagc tcgatatcga agatgagcaa
     2101 gaccgattgg agtgtcttcg gcaacgagcg gacggagttg cgcgagacat tggagagctt
     2161 caacccatcg ctaccgaggt tcggggctgg gaagctgcgg gaaaagctga gcgcggcgca
     2221 atcctcgata acatcgttgt taagtgcgat ggactcgcga gccgcatcag agcaggagtt
     2281 gcgggaatgc gaatcaaagt ggaagagctt agaagtcgaa tatcgcgccc gctcgaatat
     2341 ctgatgcgca gggaacagcc aaggcaacag agccttaatg acgtgctgcg cgatgcgcag
     2401 cgggcggcgg atgcgtggaa ccgcgaccac gctgcaccgc agaggacgta tcgcgggcgg
     2461 gctagatgat gtgttatagt agtacctgta atacacgaca aggaggtgcg acgcgaaaaa
     2521 accccgctga agcggggttt tttgcggagc ttcgcttcca cgatgcgcat gattcggagg
     2581 ccccggcgat cgccggggcc tccgtgcgtt gtgtgtcgat ttgggatgca gccagatggg
     2641 cttcctcggc gcgaaccggc cgagatttcc gaatgcagac ggtctcgctt ggacgatgcg
     2701 ctttcgaata atcccggcgg agcgggcatt ccagtcttct tatcaatata tctatagaat
     2761 gggcaaagca taaaaacttg catggactaa tgcttgaaac ccaggacaat aaccttatag
     2821 cttgtaaatt ctatcataat tgtggtttca aaatcggctc cgtcgatact atgttatacg
     2881 ccaactttga aaacaacttt gaaaaagctg ttttctggta tttaaggttt tagaatgcaa
     2941 ggaacagtga attggagttc gtcttgttat aattagcttc ttggggtatc tttaaatact
     3001 gtagaaaaga ggaaggaaat aataaatggc taaaatgaga atatcaccgg aattgaaaaa
     3061 actgatcgaa aaataccgct gcgtaaaaga tacggaagga atgtctcctg ctaaggtata
     3121 taagctggtg ggagaaaatg aaaacctata tttaaaaatg acggacagcc ggtataaagg
     3181 gaccacctat gatgtggaac gggaaaagga catgatgcta tggctggaag gaaagctgcc
     3241 tgttccaaag gtcctgcact ttgaacggca tgatggctgg agcaatctgc tcatgagtga
     3301 ggccgatggc gtcctttgct cggaagagta tgaagatgaa caaagccctg aaaagattat
     3361 cgagctgtat gcggagtgca tcaggctctt tcactccatc gacatatcgg attgtcccta
     3421 tacgaatagc ttagacagcc gcttagccga attggattac ttactgaata acgatctggc
     3481 cgatgtggat tgcgaaaact gggaagaaga cactccattt aaagatccgc gcgagctgta
     3541 tgatttttta aagacggaaa agcccgaaga ggaacttgtc ttttcccacg gcgacctggg
     3601 agacagcaac atctttgtga aagatggcaa agtaagtggc tttattgatc ttgggagaag
     3661 cggcagggcg gacaagtggt atgacattgc cttctgcgtc cggtcgatca gggaggatat
     3721 cggggaagaa cagtatgtcg agctattttt tgacttactg gggatcaagc ctgattggga
     3781 gaaaataaaa tattatattt tactggatga attgttttag atcgttaagg gatcaacttt
     3841 gggagagagt tcaaaattga tccttttttt ataacaggaa ttcttgaaga cgaaagggcc
     3901 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3961 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4021 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4081 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4141 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4201 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4261 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4321 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4381 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4441 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4501 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4561 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4621 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4681 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4741 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4801 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4861 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4921 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4981 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5041 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5101 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5701 tatggaaaaa cgccagcaac gcggcccgag atgcgccgcg tgcggctgct ggagatggcg
     5761 gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat
     5821 tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcatgagctc
     5881 ctctagtaga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac
     5941 tgagcctttc gttttatttg aaaaagaaaa gccccagcaa gcttggc
//
