LOCUS       sequence-463820-       10020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Gordonibacter pamelaeae 3C (–)-Eddh 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 10020)
  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 10020)
  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: pXD70EN25-Pct5-(–)-Eddh(Gp3C)
FEATURES             Location/Qualifiers
     source          1..10020
                     /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      6047..6074
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6197..6991)
                     /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 tggagtacaa caacgagtac aaccttgacc
     3301 tgtacccgaa gaagctgcac tggaaggagg gcgacctcac tgccacgcgc acgaccatgt
     3361 ggtcgggccc gggctgccac gagggatgcc aggtgatctt ctacacggac gacgacggca
     3421 agctcgtgaa ggttgagggc gatccgaaca gcccgttcaa ccaggggcgc ctctgcatgc
     3481 gctgcctgga gctgcccgag ctcgtgaacc acgaggagcg cctgcgctac cccatgaagc
     3541 gcgccgggaa gcgcggcgag gacaagtggg agcgcgtgag ctgggacaag gcctacgata
     3601 tcatcgagga aaaggtccgc gcgttccagg agaaggacgg ccccgagtcc atcatctcca
     3661 tgatcggcac cggccgcaac gtctcgcagg tcatcgcgca caaccagtac gcgaacttcg
     3721 gcggccccga cctcacgctc tgcttcctgt cgggcgactc gtgcatgctg ccgcgcacgg
     3781 cgctctgcta cgtggtcatg ggcaaccagt gggtggccga catgagccag ttccgccccg
     3841 accgctacga gaacgacccc gagtggcaga tccccgagtg catcgtcatc tggggcacga
     3901 acccggtcgt gtgcaactcc gacgccttcc tcggccactg gatcgtggag gccatgaagc
     3961 gcggcagcga gctcgtgacc atcgacccgc agctcacgtg gatcgcctcc aagtccaagt
     4021 actggctgcg cctgcgcccc ggcaccgacg cggcgctcgc cctcggcatg atgaacgtca
     4081 tcatcggcga ggacctcgtg gaccacgact tcatcgacaa gtggacctac ggctttgagg
     4141 cgcttgccga gcgcgtgaag gactatccgg ttgagaaggt ggccgagatc tgctgggtcg
     4201 agcccgagct catcgtggag gccgcgcgct tctacgcgaa ggcccacccc tccaccatcc
     4261 agtggggcct cgccgtggac atgtcgaaga tcggcacccc tacggcccac gccatcgcct
     4321 gcttggccgg catcacgggc aacatcgaca accccggcgg caacatcctc atcgaccagg
     4381 cctgcggcct ggagttcggc tacaactccg gcatcgagta cctcaaagag ggcatggtgg
     4441 agaagcgcct gggcaactcc aagtacaacc tgaagaagta cggcttcacg gcgtcctccc
     4501 agtccgacgc catcctcgag gcgtgcgaga ccggccttga cgagaagggc gacccgcgcc
     4561 cggtgaacat gctgtggatc cagtcctcca accccatcac gaacatgggc caggacgcgc
     4621 cgcgcatcta ccgcgccatg gagaaggtac cgttcatctg cgtggtcgac ctgttcaaga
     4681 cgccgttcat cgtggcctgc gccgacctcg tgctgccctg cgccatgtcc aacgagcgcg
     4741 actgcatccg cgtgtggttc gacccggtcc gctcgctcac gaagtgctcg agcttctacg
     4801 aggcgaagga agacgagcag atcatgatcg acctggggca ccgtttggca cccgagagct
     4861 ggcccgagtg ggtcaaggag ccgcgcgact acatgaactg gcgcctccag gtgggcaact
     4921 gcggcttcac catggaggag ctggagaacg accatgccgg catgatctac aacgacttcc
     4981 gctactacaa gtacgagaag ggcctgctgc gtcccgacgg ccagcccggc ttcatgaccc
     5041 ctacgggccg ctacgagttc tacatcagcc tgttcgacag ctggggcgtg gacgcgctgc
     5101 cgttctacga ggagcctccc acctcgccgg tctccacgcc ggagctggcc gaggagtacc
     5161 cgctcgtgct caccacgggc aagcgctcct tcgagttctt ccactccgag tggcgccagg
     5221 cgaacacgac gagccgcgag ctgcacccgg tgccctactt cgacatccat cccgacacgg
     5281 cggcgaaata cggcgtggcc gagggccagt ggacctggat cgagaaccag atgggcaagt
     5341 gccgccaggt ggcgcgcttc aacgacacgc tcgatccccg cgtcatctcc accgagcacg
     5401 gctggtggtt ccccgagcag gaggcggccg agccgagcct gttcggcgtg ttcgactgca
     5461 acccgaacaa cctcattccc atgtgcgaga acggcccgtc cggctacggc gcgcccatca
     5521 agtgcggcat ggcgaaggtg tacccctgca cgccggacaa catgaggccc gaggaccagc
     5581 ccacctacca ggtgacggcc acgaacggct acacgcacgg cccctcgcac aagagcgacg
     5641 agcctagctt ctacgaccag aacaagtaag ggggagacat catggcacaa accggcgaat
     5701 acggccttct gatcgactac cagtactgct cgggctgcca cgcctgcgag gtggcctgca
     5761 agaaggagca tgacctgccc aagggcgact tcggcatcaa gatcgtgcag gacggcccgc
     5821 gccagtgctc cgacggcgtg tgggagtacg actacctgcc cgtgcccacg cacctgtgcg
     5881 acctgtgcgc cgtgcgcgtg gaggccggca agcttcccac ctgcgtgcac cactgccagg
     5941 ccggcgtcat ggtctacggc acgctggagg agctcgccgc caaggctaag gcggccgcga
     6001 aggacaagat ggtgatcttc gcgaagtaga ctagtcgact cacactggct caccttcggg
     6061 tgggcctttc tgcgtttata tactagagag agaatataaa aagccagatt attaatccgg
     6121 cttttttatt atttaattcc tgttataaaa aaaggatcaa ttttgaactc tctcccaaag
     6181 ttgatccctt aacgatctaa aacaattcat ccagtaaaat ataatatttt attttctccc
     6241 aatcaggctt gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat
     6301 cctccctgat cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc
     6361 tcccaagatc aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca
     6421 ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca
     6481 gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca
     6541 gatcgttatt cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg
     6601 gacaatccga tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa
     6661 tcttttcagg gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac
     6721 tcatgagcag attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca
     6781 gctttccttc cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt
     6841 tataccggct gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata
     6901 ccttagcagg agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt
     6961 tcaattccgg tgatattctc attttagcca tttattattt ccttcctctt ttctacagta
     7021 tttaaagata ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc
     7081 attctaaaac cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta
     7141 taacatagta tcgacggagc cgattttgaa accacaatta tgatagaatt tacaagctat
     7201 aaggttattg tcctgggttt caagcattag tccatgcaag tttttatgct ttgcccattc
     7261 tatagatata ttgataagaa gactggaatg cccgctccgc cgggattatt cgaaagcgca
     7321 tcgtccaagc gagaccgtct gcattcggaa atctcggccg gttcgcgccg aggaagccca
     7381 tctggctgca tcccaaatcg acacacaacg cacggaggcc ccggcgatcg ccggggcctc
     7441 cgaatcatgc gcatcgtgga agcgaagctc cgcaaaaaac cccgcttcag cggggttttt
     7501 tcgcgtcgca cctccttgtc gtgtattaca ggtactacta taacacatca tctagcccgc
     7561 ccgcgatacg tcctctgcgg tgcagcgtgg tcgcggttcc acgcatccgc cgcccgctgc
     7621 gcatcgcgca gcacgtcatt aaggctctgt tgccttggct gttccctgcg catcagatat
     7681 tcgagcgggc gcgatattcg acttctaagc tcttccactt tgattcgcat tcccgcaact
     7741 cctgctctga tgcggctcgc gagtccatcg cacttaacaa cgatgttatc gaggattgcg
     7801 ccgcgctcag cttttcccgc agcttcccag ccccgaacct cggtagcgat gggttgaagc
     7861 tctccaatgt ctcgcgcaac tccgtccgct cgttgccgaa gacactccaa tcggtcttgc
     7921 tcatcttcga tatcgagctt cacgtcctcg agctggctgt caagctcggc aaggtcaccc
     7981 tccttgtcgg caatctcggt aacaagctct gccttacgcg tttcggcatc attcagtaaa
     8041 gcggcggctt cttccctagc cgccgcgacg attcgcgccg cttcgtcctt cgcaggttca
     8101 accacggcct tatcgaccgc acagcgtacc ttgtcgatat ccacggactt gtccgtgtat
     8161 acacgctgcg cccgcgtctc ctcgtcaagc tcgattgcag ggcggtagcc catgcgcccc
     8221 tctaggtaat ccccaagctc cttgtgcatg ctccggtaga agctgcgggg gcacatctgc
     8281 ttgtagttga accgtccatc taggatgggc gtgaacggca cgtgcatgtg gtcgcgggcg
     8341 ggcgtgccat ccttcaacac ctcgtccttg tggacgtacc cgcccatcat gttctcgatg
     8401 ccgaacttct tcgacaggta ccagtacgtg agacggaaga acctgtcctc atcgcccttg
     8461 cgcacattgt cgggaagcgt cacaacaacg tcagccataa cgaccgcatc cttgcgcgtg
     8521 gcacgcttcc ctgcggcctt ctgggcctcg ttcacgcgct cgatgcggct ctccaccgtg
     8581 ccccagttcg gctcgccatc gcgcggctcc acgcgcccca caacgagctt gccgtccttg
     8641 ttcgtgtagt ggccgaccac caagtcgttc ttgatgcgct cggggtcgat gttgtcgcgc
     8701 tccaggctgc tgcggtcgcg gcggtagtgg gcaagcatgt gcccgaccga tggagccttg
     8761 tatttcgcga tgtgagccat ggaaacctcc gaaccgtaca gacttaatcg actaaagtct
     8821 agcacgttag accttagcta cgctaaggac gtggcagggc cagccctgca cccgctcggg
     8881 cgttcccccc gagacccccc agcccttgcc cgttcgcggc ttcgctcacg gccctcgggc
     8941 catcggcgcg agcgccgaac ggagggactt cgcccctccg aaccacccct caatctacct
     9001 gcggttcggc acacggcaca cggcactatt aggggggccg tcgtagtggc ccccctgtgg
     9061 tcatttgtca tttgtcattt gtcacattat ttttcggaca cattattttt cggacgtagt
     9121 tagcgaaaaa taatgagccg cccgattctg caggtctcat ccagtcgaga cacgggcaag
     9181 gaaagggtcg atggagcaga gcgtccatcg acccttaatc aagtctggtc tattcggttg
     9241 gccgtcgcgt caattccgca atccgtgaac gagtcggctt agatctgcgc gagcgcagac
     9301 cacgccgccc cgtagtcacg cattgctgca ttgacccgac tactctttgg cacctgactg
     9361 ttccacgcac ttgcgcttac gttcggcgtt cagatagttc ccgcgacaat agcgctcgag
     9421 aatcgagttg taccccacga tgatttcgcg gtactgctcg ggaaactccc tcagcgccgc
     9481 agccgtgaaa tcgttgatgg tctcgaacct ctcgcacatg atgaagccga ccaactcgaa
     9541 gagcatgtcc tgcttgtcct tagtcgagag ctgttcgagc tgaccaatgt cgaagttgtt
     9601 tccctcgatg cgtgcttcga ccggatacag ctccttgccc ttcttgcgac aggaagccgt
     9661 gtcgtgaatc agatagtcat acgcatgcct gatgttgttg attggctcgg ctgacgaaca
     9721 gcacttcttg ccgtccgcgc tcaatcggtt caagacgttg atgattgcct tgcgtgtcgt
     9781 gttgccgccc cagacaacaa tgacgtgcgc atgcgtcttg cgctgcttgc tcttctggtc
     9841 atggtctatg tcgtgaaccg cataagcgaa aggcacttga agaatgtcgt ctatctcgtc
     9901 ctgccagtca tcgcgcatgt tctctgggta gcagatgccc tcccagagct tcaccttcgt
     9961 ctcgccagcg tttgccgcca tgctataatc tccttgtctc gaatgcccct gctgccgcca
//
