LOCUS       sequence-463796-        9956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Gordonibacter pamelaeae DSM 19378 (+)-Eldh1 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 9956)
  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 9956)
  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: pXD70EN20-Pct5-GPA_25810-(+)-Eldh1(GpDSM19378)
FEATURES             Location/Qualifiers
     source          1..9956
                     /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      5983..6010
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6133..6927)
                     /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 tgtcgaacga ttactacgaa agccaccgag
     3301 gcgaatggca gtggcaggac ggcgagtaca ccgtcacgcg cacgaccgcc tggtcggggc
     3361 ccggttgcca cgacggctgc ggcgtgctgt actacacgaa ggacggcaag ctggagaagg
     3421 tggagggcga ccccgacaac ccgttcaacc agggcgcgct gtgcatgcgc tgcctggaga
     3481 tgcccgagtt cgtgaacaac cccacgcgcc tcggccaccc catgaagcgt gccggcgagc
     3541 ggggcgagaa caaatgggag cgcgtgtcct gggaggaggc gctcgacctc atcgaggaaa
     3601 aggtacgcgc catctggaag gactacggcc ccgagtccat cgtctgtcat atcggcaccg
     3661 ggcgcaacaa ctgcgaccag ataccgtacc tgtgctacgc ggcgttcgga tcgccgaact
     3721 tctccatggg ctatctggcc ggcgactcct gcttctgccc gcgctccgcg tcgatggcgt
     3781 cgcagaacgg cgacttcatg atcgccgact gctcgcagca gttcgagaag cgctacgacg
     3841 agagcacggg ctggcgctgc cccgaagtca tcgtgaactg gggctgcaac gcggtcgtct
     3901 cgaactccga caactttttc ggcggctgga tcgtggactg catgcagcgc ggctccaaga
     3961 tgatcaccat cgacccgtcg ctcacctggc tggcctcccg ctcggcgtac tggctgcgcg
     4021 tgcggccggg caccgacgcg gctttggcca tggccatgct caacgtcatc atctccgagg
     4081 acctctacga ccacgatttc gtggccaagt ggtgctacgg cttcgacgag cttgccgagc
     4141 gcgtgcgcga ctggacgccc gagcgggctg cggaggtttg ctgggtcgag gccgaggaca
     4201 tcgtgggcgc cgcccgcctg tacgcccagg ccgaccccgc cgccatccag tgggggctca
     4261 agctcgacca gaccaccaac gccaccgcca ccgcgtgcgc cgtcaactcg ctgtgggcca
     4321 tcacgggcaa cgtggacaat cccggcggca acatcatcat ccgcaacgcc ttcgaccaga
     4381 acctctcgta cggctacggg taccacctgc tgccgcccga ggtgcaggcg aagcgcctgg
     4441 gggccgagtt cccgctgatg agccgtgcgg gctacagctc ctccgcccat gcggactccg
     4501 tgctgaaggc catcgagacc ggcgagccgt atcctatccg catggtgtgg atgacgtcca
     4561 cgaaccccat cgccaacatg ggcgccgacg cgccgcgcct gttccgcgcc ttgaagagcg
     4621 tcgatttcgt ggtggtgaac gacctgttca tgacgccaac cgccatcgcc ttcgccgatc
     4681 tggtgctgcc cgccgccatg tctcccgaac gcaactcgca gcgctgctgg tgggttccga
     4741 accgcacgct caccaaggtc acccagtacg aggagtgcgt ttcggacgac gacctggtca
     4801 ccatggtggg caaacgcctg caccccgaga acttcccctg ggacgacaac gtgggatgga
     4861 tcaaccacat cttcgagcac gagacgaacg acgagctgcc ctacaagagc ttcgaggagg
     4921 cgcagaaggt cgtgtggtcc tacccggcgt tcacctacct caagcacgag aagggcctgc
     4981 tgcgccccga cgggcagccc gggttcaaca cgccgtccgg tcgcatcgag ctgtacaaca
     5041 cgaacctcat gctgtggggc tacgatccgc tgcccgactt caaggagccc agctgctctc
     5101 cggtgagcag ccccgagctc gccgagaagt acccgctcgt cctgaccacc ggcgcgcggt
     5161 ccttcgagtt cttccattcc gagcatcgcc aggagggcac gacgtcgcgc gagctgcacc
     5221 ccgacccgct gttcgagatg agccctgccg ccgccgaggc cgcgggcctc tccgagggcg
     5281 attgggcctg gatcgagaac cagcgcggcc gctgccgcca gaagctgcac gtgaacccct
     5341 cgctcgacga ccgcgtggtg cgcgccgagc acggctggtg gttccccgag accgaggggg
     5401 cggagccgaa cctgtacggc gtgttcgact ccaacatcaa caacctcacc ccgcagtgcg
     5461 agaacggcga caccggcttc ggcgccccgt acgcgaacca gctgtgccgc gtgtacccgt
     5521 gcactgagga gaacagccgt gtcctgccgg gcgagcaggt caccaggcgc gggggctatt
     5581 ccaaggtaat cgacgagtcc aacggcgacg tgaaggtggg tgctatctat ggctaacgca
     5641 ttgatgatcg actacgagtt ctgcacgggc tgccacgcct gcgaggtggc ctgcaagaag
     5701 catctgggcc tgcccgcggg ccagcacggc atccgcgtgc tccaggacgg cccccgcgag
     5761 ctcgcgggcg gagggtggga gtacaacttc ctgcccatgc ccacgtcgct ctgcgacctg
     5821 tgcgcctccc gcctggccga gggcaaggac gccgcctgcg tccaccactg cccggcccac
     5881 tgcatggcgc tcgggccggc cgaggagctg gcggcacggt tcgacgggaa ggtcaaccgc
     5941 gcactgttcg ttcccgcgca gtaaaactag tcgactcaca ctggctcacc ttcgggtggg
     6001 cctttctgcg tttatatact agagagagaa tataaaaagc cagattatta atccggcttt
     6061 tttattattt aattcctgtt ataaaaaaag gatcaatttt gaactctctc ccaaagttga
     6121 tcccttaacg atctaaaaca attcatccag taaaatataa tattttattt tctcccaatc
     6181 aggcttgatc cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc
     6241 cctgatcgac cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc
     6301 aagatcaata aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc
     6361 gccgtgggaa aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc
     6421 gcgcggatct ttaaatggag tgtcttcttc ccagttttcg caatccacat cggccagatc
     6481 gttattcagt aagtaatcca attcggctaa gcggctgtct aagctattcg tatagggaca
     6541 atccgatatg tcgatggagt gaaagagcct gatgcactcc gcatacagct cgataatctt
     6601 ttcagggctt tgttcatctt catactcttc cgagcaaagg acgccatcgg cctcactcat
     6661 gagcagattg ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt
     6721 tccttccagc catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata
     6781 ccggctgtcc gtcattttta aatataggtt ttcattttct cccaccagct tatatacctt
     6841 agcaggagac attccttccg tatcttttac gcagcggtat ttttcgatca gttttttcaa
     6901 ttccggtgat attctcattt tagccattta ttatttcctt cctcttttct acagtattta
     6961 aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt ccttgcattc
     7021 taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt ggcgtataac
     7081 atagtatcga cggagccgat tttgaaacca caattatgat agaatttaca agctataagg
     7141 ttattgtcct gggtttcaag cattagtcca tgcaagtttt tatgctttgc ccattctata
     7201 gatatattga taagaagact ggaatgcccg ctccgccggg attattcgaa agcgcatcgt
     7261 ccaagcgaga ccgtctgcat tcggaaatct cggccggttc gcgccgagga agcccatctg
     7321 gctgcatccc aaatcgacac acaacgcacg gaggccccgg cgatcgccgg ggcctccgaa
     7381 tcatgcgcat cgtggaagcg aagctccgca aaaaaccccg cttcagcggg gttttttcgc
     7441 gtcgcacctc cttgtcgtgt attacaggta ctactataac acatcatcta gcccgcccgc
     7501 gatacgtcct ctgcggtgca gcgtggtcgc ggttccacgc atccgccgcc cgctgcgcat
     7561 cgcgcagcac gtcattaagg ctctgttgcc ttggctgttc cctgcgcatc agatattcga
     7621 gcgggcgcga tattcgactt ctaagctctt ccactttgat tcgcattccc gcaactcctg
     7681 ctctgatgcg gctcgcgagt ccatcgcact taacaacgat gttatcgagg attgcgccgc
     7741 gctcagcttt tcccgcagct tcccagcccc gaacctcggt agcgatgggt tgaagctctc
     7801 caatgtctcg cgcaactccg tccgctcgtt gccgaagaca ctccaatcgg tcttgctcat
     7861 cttcgatatc gagcttcacg tcctcgagct ggctgtcaag ctcggcaagg tcaccctcct
     7921 tgtcggcaat ctcggtaaca agctctgcct tacgcgtttc ggcatcattc agtaaagcgg
     7981 cggcttcttc cctagccgcc gcgacgattc gcgccgcttc gtccttcgca ggttcaacca
     8041 cggccttatc gaccgcacag cgtaccttgt cgatatccac ggacttgtcc gtgtatacac
     8101 gctgcgcccg cgtctcctcg tcaagctcga ttgcagggcg gtagcccatg cgcccctcta
     8161 ggtaatcccc aagctccttg tgcatgctcc ggtagaagct gcgggggcac atctgcttgt
     8221 agttgaaccg tccatctagg atgggcgtga acggcacgtg catgtggtcg cgggcgggcg
     8281 tgccatcctt caacacctcg tccttgtgga cgtacccgcc catcatgttc tcgatgccga
     8341 acttcttcga caggtaccag tacgtgagac ggaagaacct gtcctcatcg cccttgcgca
     8401 cattgtcggg aagcgtcaca acaacgtcag ccataacgac cgcatccttg cgcgtggcac
     8461 gcttccctgc ggccttctgg gcctcgttca cgcgctcgat gcggctctcc accgtgcccc
     8521 agttcggctc gccatcgcgc ggctccacgc gccccacaac gagcttgccg tccttgttcg
     8581 tgtagtggcc gaccaccaag tcgttcttga tgcgctcggg gtcgatgttg tcgcgctcca
     8641 ggctgctgcg gtcgcggcgg tagtgggcaa gcatgtgccc gaccgatgga gccttgtatt
     8701 tcgcgatgtg agccatggaa acctccgaac cgtacagact taatcgacta aagtctagca
     8761 cgttagacct tagctacgct aaggacgtgg cagggccagc cctgcacccg ctcgggcgtt
     8821 ccccccgaga ccccccagcc cttgcccgtt cgcggcttcg ctcacggccc tcgggccatc
     8881 ggcgcgagcg ccgaacggag ggacttcgcc cctccgaacc acccctcaat ctacctgcgg
     8941 ttcggcacac ggcacacggc actattaggg gggccgtcgt agtggccccc ctgtggtcat
     9001 ttgtcatttg tcatttgtca cattattttt cggacacatt atttttcgga cgtagttagc
     9061 gaaaaataat gagccgcccg attctgcagg tctcatccag tcgagacacg ggcaaggaaa
     9121 gggtcgatgg agcagagcgt ccatcgaccc ttaatcaagt ctggtctatt cggttggccg
     9181 tcgcgtcaat tccgcaatcc gtgaacgagt cggcttagat ctgcgcgagc gcagaccacg
     9241 ccgccccgta gtcacgcatt gctgcattga cccgactact ctttggcacc tgactgttcc
     9301 acgcacttgc gcttacgttc ggcgttcaga tagttcccgc gacaatagcg ctcgagaatc
     9361 gagttgtacc ccacgatgat ttcgcggtac tgctcgggaa actccctcag cgccgcagcc
     9421 gtgaaatcgt tgatggtctc gaacctctcg cacatgatga agccgaccaa ctcgaagagc
     9481 atgtcctgct tgtccttagt cgagagctgt tcgagctgac caatgtcgaa gttgtttccc
     9541 tcgatgcgtg cttcgaccgg atacagctcc ttgcccttct tgcgacagga agccgtgtcg
     9601 tgaatcagat agtcatacgc atgcctgatg ttgttgattg gctcggctga cgaacagcac
     9661 ttcttgccgt ccgcgctcaa tcggttcaag acgttgatga ttgccttgcg tgtcgtgttg
     9721 ccgccccaga caacaatgac gtgcgcatgc gtcttgcgct gcttgctctt ctggtcatgg
     9781 tctatgtcgt gaaccgcata agcgaaaggc acttgaagaa tgtcgtctat ctcgtcctgc
     9841 cagtcatcgc gcatgttctc tgggtagcag atgccctccc agagcttcac cttcgtctcg
     9901 ccagcgtttg ccgccatgct ataatctcct tgtctcgaat gcccctgctg ccgcca
//
