LOCUS       sequence-266458-        3234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible expression of R-TAGA-Evo1 qtRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3234)
  AUTHORS   DeBenedictis EA, Carver GD, Chung CZ, Soll D, Badran AH
  TITLE     Multiplex suppression of four quadruplet codons via tRNA directed 
            evolution.
  JOURNAL   Nat Commun. 2021 Sep 29;12(1):5706. doi: 10.1038/s41467-021-25948-y.
  PUBMED    34588441
REFERENCE   2  (bases 1 to 3234)
  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: "2021-09-29"; volume: "12(1):5706. doi"; pages: " 
            10.1038/s41467-021-25948-y"
COMMENT     Sequence Label: pED14xR-3
FEATURES             Location/Qualifiers
     source          1..3234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        550..584
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     CDS             611..1759
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     promoter        1769..1860
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1861..2652
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     protein_bind    2941..2957
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 cacttgaggg gccgactcac ccggcgcggc gttgacagat gaggggcagg ctcgatttcg
       61 gccggcgacg tggagctggc cagcctcgca aatcggcgaa aacgcctgat tttacgcgag
      121 tttcccacag atgatgtgga caagcctggg gataagtgcc ctgcggtatt gacacttgag
      181 gggcgcgact actgacagat gaggggcgcg atccttgaca cttgaggggc agagtgctga
      241 cagatgaggg gcgcacctat tgacatttga ggggctgtcc acaggcagaa aatccagcat
      301 ttgcaagggt ttccgcccgt ttttcggcca ccgctaacct gtcttttaac ctgcttttaa
      361 accaatattt ataaaccttg tttttaacca gggctgcgcc ctgtgcgcgt gaccgcgcac
      421 gccgaagggg ggtgcccccc cttctcgaac cctcccggcc cgctaacgcg ggcctcccat
      481 ccccccaggg gctgcgcccc tcggccgcga acggcctcac cccaaaaatg gaggtttctc
      541 ttctttcact ttacggctag ctcagcccta ggtattatgc tagcagcccc atacgatata
      601 aggagggtga atgaatcgga cgtttgaccg gaaggcatac aggcaagaac tgatcgacgc
      661 ggggttttcc gccgaggatg ccgaaaccat cgcaagccgc accgtcatgc gtgcgccccg
      721 cgaaaccttc cagtccgtcg gctcgatggt ccagcaagct acggccaaga tcgagcgcga
      781 cagcgtgcaa ctggctcccc ctgccctgcc cgcgccatcg gccgccgtgg agcgttcgcg
      841 tcgtctcgaa caggaggcgg caggtttggc gaagtcgatg accatcgaca cgcgaggaac
      901 tatgacgacc aagaagcgaa aaaccgccgg cgaggacctg gcaaaacagg tcagcgaggc
      961 caagcaggcc gcgttgctga aacacacgaa gcagcagatc aaggaaatgc agctttcctt
     1021 gttcgatatt gcgccgtggc cggacacgat gcgagcgatg ccaaacgaca cggcccgctc
     1081 tgccctgttc accacgcgca acaagaaaat cccgcgcgag gcgctgcaaa acaaggtcat
     1141 tttccacgtc aacaaggacg tgaagatcac ctacaccggc gtcgagctgc gggccgacga
     1201 tgacgaactg gtgtggcagc aggtgttgga gtacgcgaag cgcaccccta tcggcgagcc
     1261 gatcaccttc acgttctacg agctttgcca ggacctgggc tggtcgatca atggccggta
     1321 ttacacgaag gccgaggaat gcctgtcgcg cctacaggcg acggcgatgg gcttcacgtc
     1381 cgaccgcgtt gggcacctgg aatcggtgtc gctgctgcac cgcttccgcg tcctggaccg
     1441 tggcaagaaa acgtcccgtt gccaggtcct gatcgacgag gaaatcgtcg tgctgtttgc
     1501 tggcgaccac tacacgaaat tcatatggga gaagtaccgc aagctgtcgc cgacggcccg
     1561 acggatgttc gactatttca gctcgcaccg ggagccgtac ccgctcaagc tggaaacctt
     1621 ccgcctcatg tgcggatcgg attccacccg cgtgaagaag tggcgcgagc aggtcggcga
     1681 agcctgcgaa gagttgcgag gcagcggcct ggtggaacac gcctgggtca atgatgacct
     1741 ggtgcattgc aaacgctaga cggcctattt ggcctatttt tctaaataca ttcaaatatg
     1801 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1861 atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
     1921 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
     1981 ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
     2041 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
     2101 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
     2161 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
     2221 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
     2281 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
     2341 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
     2401 ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
     2461 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
     2521 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
     2581 tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta
     2641 gtcggcaaat aaacgccatg ggcattaaaa aatgccctct tgggttatca agagggtcat
     2701 tatatttgct gaggagtgcc gttaattaag tcggatgaat ggcagaagct taaaaaaaat
     2761 ccttagcttt cgctaaggat ctgcaggtcg acggatcctg ctgaaggtta cggagaatat
     2821 ggtgcatccg ggaggattcg aacctccgac cgatcggttt agaagccgag tactctatcc
     2881 agctgagcta cggatgcatc gggaaaactt attctacaag ggacccatct tgtctgctaa
     2941 ttgttatccg ctcacaatta tcttactgcg cagatacgcc ctcgtcaatc ccttaatagc
     3001 aaaatgcctt ttgatcggcg agaaagtcag caggccgctt agttagccgt gcactgatca
     3061 tatcgtcaat tattacctcc acggggagag cctgagcaaa ctggcctcag gcatttgaga
     3121 agcacaaggg tccctaagtc tcctcagcaa aacgaaaggc ccagtctttc gactgagcct
     3181 ttcgttttat ttgaccggat gtcctcttac tgacagatga ggggcggacg ttga
//
