LOCUS       sequence-463827-        9905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Gordonibacter pamelaeae DSM 19378 (+)-Eldh2 in Eggerthella
            lenta DSM 2243 with a cumate inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9905)
  AUTHORS   Dong X, Bae M, Le C, Aguilar Ramos MA, Balskus EP
  TITLE     Enantiocomplementary Gut Bacterial Enzymes Metabolize Dietary 
            Polyphenols.
  JOURNAL   J Am Chem Soc. 2025 Mar 5;147(9):7231-7244. doi: 
            10.1021/jacs.4c09892. Epub 2025 Feb 24.
  PUBMED    39993729
REFERENCE   2  (bases 1 to 9905)
  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"; doi: 
            "10.1021/jacs.4c09892"; journalName: "J Am Chem Soc"; date: 
            "2025-03-5- 5"; volume: "147"; issue: "9"; pages: "7231-7244"
COMMENT     Sequence Label: pXD70EN21-Pct5-GPA_25800-(+)-Eldh2(GpDSM19378)
FEATURES             Location/Qualifiers
     source          1..9905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      24..95
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(276..864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(356..375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1035..1826,1827..1895))
                     /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     1658..1677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1896..2000)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2068..2086
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2243..2262
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     CDS             complement(2265..2876)
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    3214..3241
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     terminator      5932..5959
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6082..6876)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 agcttgctgg ggcttttctt tttcaaataa aacgaaaggc tcagtcgaaa gactgggcct
       61 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gaggagctca tgccggcggc
      121 acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg
      181 tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc
      241 gcacgcggcg catctcgggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
      721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     1081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     1141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     1201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     1261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     1321 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     1381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     1501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1621 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     1681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     2041 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
     2101 ggcgaaaaaa ccccgccgaa gcggggtttt ttgcgaaata taatgaccct cttgataacc
     2161 caagagggca ttttttagga acggttattt gattattaat ccggcggcaa ctgaaacgat
     2221 tcggatcctg tattactatt ctgaattcgc ggccgcttct agatctagcg cttgaatttc
     2281 gcgtaccgct ctcgcgcaat ttcgagtgtc gagttcctga cacgctcaaa gcgttctttg
     2341 tccttctgcc ataggctacg aacagcaagc ccacgcaccg aattgaatat caaccaaagg
     2401 atatcttctg catcatcacg cgaaagacca cggctcacca gaacaccaag ccacatatcc
     2461 tcgacgacaa accgattcct ctctaccgtg cgctgaatac cctcgcgtaa cgctgggtcc
     2521 cggtcggcag ccacaatcaa atcaaggctg atagagaagt catcgtcgag gaaaaattcg
     2581 gcggcgtcgt ccagcatttg ctggatgacg tcatcctctg gcttcaattt cgctaatcga
     2641 gcccgactgc gttcggtgat ctgttcgtaa agccattcaa aagtggcaag cagaagctca
     2701 agctttgtcg ggaaatgatg gctctgcgct cctctggaga caccagcagc accgggcaca
     2761 tctgcgatcc ggaatcccgc gtaacctttt tcccgtaaaa cccccagggc cgctgcaatc
     2821 aacttgccct gggtctccat tgcgcgctct gcctgtgttc ttctctttgg actcatgatc
     2881 accatttttt ttactaaagc ttgatctgca atttgaataa taaccactcc tttgtttatc
     2941 caccgaacta agttggtgtt ttttgaagct tgaattagat atttaaaagt atcatatcta
     3001 atattataac taaattttct aaaaaaaaca ttgaaggaac ggttatttga ttattaatcc
     3061 ggctttttta ttatttaggc aactgaaacg attcggatcc tgtattacta ttctattttg
     3121 tcaaaataat tttattgaca acgtcttatt aacgttgata taatttaaat tttatttgac
     3181 aaaaatgggc tcgtgttgta caataaatgt agtaacaaac agacaatctg gtctgtttgt
     3241 attatgatta actaataagg aggacaaaca tggatcaggc caatagctac tacgaggccc
     3301 ataagggcga ggtggagtgg cgcgacggcg actacacggt gacgcgcacg acggtgtggt
     3361 cggggcccgg ttgccacgac ggctgcggcg tgctgtacta cacgaaggac ggcaagctgg
     3421 agaaggtgga gggcgacccc gactgcccct tcaaccaggg cacgctgtgc atgcgctgcc
     3481 tggagatgcc cgaagccgtg aaccatcccg accggccgac cggtccgctc aagcgggcgg
     3541 gcgagcgcgg cgagaacaag tgggagtccg tcacctggga cgaggcctac gacattatcg
     3601 aggagaaggt gcgcgccatc tggaaggact acggcgccga gtccatcgcg tgcatgatcg
     3661 gcaccggtcg caacaactgc tggcagatcc cgtacctgtg ctatgcggcg ttcggctcgc
     3721 cgaacttctg cctgggattc ctctcgggcg agtcgtgctt ccagccacgc tcggcggcca
     3781 tggcctccat gaacggcgac ttcctcatcg ccgacatgtc acagcagttc gagagccgct
     3841 acgacaccga gaacaccgag tgggttccgc ctgagatgat cgtggtctgg gggtgcaacc
     3901 ccatcgcctc gaactccgac aacttctacg gaggctgggt cgtcgactgc atgcagcgcg
     3961 gcagccggtt gatcaccgtc gacccgtcgc tcacctggtt ggcggcccgt tccgactatt
     4021 ggctgcgcct gcgccccggc accgacggcg cgctcgccct ggccatgctc aacgtcatca
     4081 tctccgagga cctctacgac cgcgagttcg tcgagaagtg gtgctacggc ttcgacgagc
     4141 ttgccgagcg cgtcaaggaa tggacgccgg agcgggcgga ggcggtgtgc tgggttccga
     4201 aggacaccat aatcggggcg gcgcgcgcct acgcgaaggc caagcccgcc accatccact
     4261 ggggcctgcc cattgaccag gcggttgtcg gcatccccac ggcgcaggcc atcaacgcgc
     4321 tgtgggccgt caccggcaac acggacgtgc cgggcggcaa catcatcatc cgcaacgcgt
     4381 tcgagcagaa cgtgtcgtac aactacggat accagaccct tcccgccgag gtgcaggcca
     4441 agcgcatcgg cgcggagtac ccgctcatgt ccaaggccgg gttcagctcc acggcccact
     4501 cggactccat cctgcaggcc atcgagaccg gcaagccgta tcccatccgc atgctctggc
     4561 tttcctccac gaaccctatc gccaacatgg cgtccgacgc gccgcgcgtc taccgggcgg
     4621 tgcgcaccct cgacttcgtc gtggtgtccg acccgttcct cacccccacg gccgtggcgt
     4681 tcgccgacgt gttcctgccg gcgtccatga gctgcgagcg caacacgcag cgcgtatggt
     4741 gggtgccgct gcgcggcatg aagaaggtga cgcagttcgc cgacgtgcgt tccgacgaca
     4801 ccatcgtcac ggacctcgga cgccgcctgc atcccgagaa cttcccctgg accgatgaca
     4861 tcggctggac gaacgatatc ctcgtcaacg acacgccagg ttacgacggc gacttcgagc
     4921 agctgtgcaa ggacgtgtac gtctatcccg cgttcgagta ccgcaagcac gagaagggcc
     4981 tgctgcgccc cgacgggcag cccggtttca acacgccgtc cggccgcatc gagctctaca
     5041 acagcatctt cgcgctgtgg ggctacgacc cgctgccgca gtgggaggag ccatcctgct
     5101 cgccgtacgc ctcgccggag ctgtacgagc agtacccctt cgtgctgacg acgggcgcgc
     5161 gctcggtgga gttcttccat tccgagcatc gccagctgga gacgagccgc gagttccacc
     5221 ccgatccgct gttcgagatg agcccgaagg ccgccgaggc cgcagggctg tccgaggggg
     5281 attggtgctg gctggagaac catcgcggcc gctgccgcca gaagctgcac gtgaacccct
     5341 cgctcgacga ccgcgtggtg cgcgccgagc acggctggtg gttcccggag caggaggccg
     5401 cggaacctgt cctgttcgga gtgttcgact ccaacatcaa caacctgctg ccgcagtgcg
     5461 agaacgggcc gaccggttac ggtgccccgt acaagaacca gctgtgccgt gtgtacgcct
     5521 gcacggagga gaacagcgcc gagctgccca cggcatgcgt gacgaggaag gacgggtact
     5581 cgaaatgagc gctaacgcat tgatgatcga ctacgggttc tgcacgggct gccacgcctg
     5641 cgaggtggcc tgcaagaagc atctgggcct gcccgcgggc cagcacggca tccgcgtgct
     5701 ccaggacggc ccccgcgagc tcgcgggcgg agggtgggag tacaacttcc tgcccatgcc
     5761 cacgtcgctc tgcgacctgt gcgcctcccg cctggccgag ggcaaggacg ccgcctgcgt
     5821 ccaccactgc ccggcccact gcatggcgct cgggccggcc gaggagctgg cggcgagggc
     5881 tgccgagaag acgcggtgcg tggtcttctc ctagactagt cgactcacac tggctcacct
     5941 tcgggtgggc ctttctgcgt ttatatacta gagagagaat ataaaaagcc agattattaa
     6001 tccggctttt ttattattta attcctgtta taaaaaaagg atcaattttg aactctctcc
     6061 caaagttgat cccttaacga tctaaaacaa ttcatccagt aaaatataat attttatttt
     6121 ctcccaatca ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc
     6181 gatatcctcc ctgatcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc
     6241 gcttctccca agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc
     6301 tcccaggtcg ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc
     6361 atacagctcg cgcggatctt taaatggagt gtcttcttcc cagttttcgc aatccacatc
     6421 ggccagatcg ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt
     6481 atagggacaa tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc
     6541 gataatcttt tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc
     6601 ctcactcatg agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac
     6661 aggcagcttt ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt
     6721 ccctttatac cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt
     6781 atatacctta gcaggagaca ttccttccgt atcttttacg cagcggtatt tttcgatcag
     6841 ttttttcaat tccggtgata ttctcatttt agccatttat tatttccttc ctcttttcta
     6901 cagtatttaa agatacccca agaagctaat tataacaaga cgaactccaa ttcactgttc
     6961 cttgcattct aaaaccttaa ataccagaaa acagcttttt caaagttgtt ttcaaagttg
     7021 gcgtataaca tagtatcgac ggagccgatt ttgaaaccac aattatgata gaatttacaa
     7081 gctataaggt tattgtcctg ggtttcaagc attagtccat gcaagttttt atgctttgcc
     7141 cattctatag atatattgat aagaagactg gaatgcccgc tccgccggga ttattcgaaa
     7201 gcgcatcgtc caagcgagac cgtctgcatt cggaaatctc ggccggttcg cgccgaggaa
     7261 gcccatctgg ctgcatccca aatcgacaca caacgcacgg aggccccggc gatcgccggg
     7321 gcctccgaat catgcgcatc gtggaagcga agctccgcaa aaaaccccgc ttcagcgggg
     7381 ttttttcgcg tcgcacctcc ttgtcgtgta ttacaggtac tactataaca catcatctag
     7441 cccgcccgcg atacgtcctc tgcggtgcag cgtggtcgcg gttccacgca tccgccgccc
     7501 gctgcgcatc gcgcagcacg tcattaaggc tctgttgcct tggctgttcc ctgcgcatca
     7561 gatattcgag cgggcgcgat attcgacttc taagctcttc cactttgatt cgcattcccg
     7621 caactcctgc tctgatgcgg ctcgcgagtc catcgcactt aacaacgatg ttatcgagga
     7681 ttgcgccgcg ctcagctttt cccgcagctt cccagccccg aacctcggta gcgatgggtt
     7741 gaagctctcc aatgtctcgc gcaactccgt ccgctcgttg ccgaagacac tccaatcggt
     7801 cttgctcatc ttcgatatcg agcttcacgt cctcgagctg gctgtcaagc tcggcaaggt
     7861 caccctcctt gtcggcaatc tcggtaacaa gctctgcctt acgcgtttcg gcatcattca
     7921 gtaaagcggc ggcttcttcc ctagccgccg cgacgattcg cgccgcttcg tccttcgcag
     7981 gttcaaccac ggccttatcg accgcacagc gtaccttgtc gatatccacg gacttgtccg
     8041 tgtatacacg ctgcgcccgc gtctcctcgt caagctcgat tgcagggcgg tagcccatgc
     8101 gcccctctag gtaatcccca agctccttgt gcatgctccg gtagaagctg cgggggcaca
     8161 tctgcttgta gttgaaccgt ccatctagga tgggcgtgaa cggcacgtgc atgtggtcgc
     8221 gggcgggcgt gccatccttc aacacctcgt ccttgtggac gtacccgccc atcatgttct
     8281 cgatgccgaa cttcttcgac aggtaccagt acgtgagacg gaagaacctg tcctcatcgc
     8341 ccttgcgcac attgtcggga agcgtcacaa caacgtcagc cataacgacc gcatccttgc
     8401 gcgtggcacg cttccctgcg gccttctggg cctcgttcac gcgctcgatg cggctctcca
     8461 ccgtgcccca gttcggctcg ccatcgcgcg gctccacgcg ccccacaacg agcttgccgt
     8521 ccttgttcgt gtagtggccg accaccaagt cgttcttgat gcgctcgggg tcgatgttgt
     8581 cgcgctccag gctgctgcgg tcgcggcggt agtgggcaag catgtgcccg accgatggag
     8641 ccttgtattt cgcgatgtga gccatggaaa cctccgaacc gtacagactt aatcgactaa
     8701 agtctagcac gttagacctt agctacgcta aggacgtggc agggccagcc ctgcacccgc
     8761 tcgggcgttc cccccgagac cccccagccc ttgcccgttc gcggcttcgc tcacggccct
     8821 cgggccatcg gcgcgagcgc cgaacggagg gacttcgccc ctccgaacca cccctcaatc
     8881 tacctgcggt tcggcacacg gcacacggca ctattagggg ggccgtcgta gtggcccccc
     8941 tgtggtcatt tgtcatttgt catttgtcac attatttttc ggacacatta tttttcggac
     9001 gtagttagcg aaaaataatg agccgcccga ttctgcaggt ctcatccagt cgagacacgg
     9061 gcaaggaaag ggtcgatgga gcagagcgtc catcgaccct taatcaagtc tggtctattc
     9121 ggttggccgt cgcgtcaatt ccgcaatccg tgaacgagtc ggcttagatc tgcgcgagcg
     9181 cagaccacgc cgccccgtag tcacgcattg ctgcattgac ccgactactc tttggcacct
     9241 gactgttcca cgcacttgcg cttacgttcg gcgttcagat agttcccgcg acaatagcgc
     9301 tcgagaatcg agttgtaccc cacgatgatt tcgcggtact gctcgggaaa ctccctcagc
     9361 gccgcagccg tgaaatcgtt gatggtctcg aacctctcgc acatgatgaa gccgaccaac
     9421 tcgaagagca tgtcctgctt gtccttagtc gagagctgtt cgagctgacc aatgtcgaag
     9481 ttgtttccct cgatgcgtgc ttcgaccgga tacagctcct tgcccttctt gcgacaggaa
     9541 gccgtgtcgt gaatcagata gtcatacgca tgcctgatgt tgttgattgg ctcggctgac
     9601 gaacagcact tcttgccgtc cgcgctcaat cggttcaaga cgttgatgat tgccttgcgt
     9661 gtcgtgttgc cgccccagac aacaatgacg tgcgcatgcg tcttgcgctg cttgctcttc
     9721 tggtcatggt ctatgtcgtg aaccgcataa gcgaaaggca cttgaagaat gtcgtctatc
     9781 tcgtcctgcc agtcatcgcg catgttctct gggtagcaga tgccctccca gagcttcacc
     9841 ttcgtctcgc cagcgtttgc cgccatgcta taatctcctt gtctcgaatg cccctgctgc
     9901 cgcca
//
