LOCUS       sequence-266435-        3240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible expression of Y-TAGA-Evo1 qtRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3240)
  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 3240)
  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: pED14xY-3
FEATURES             Location/Qualifiers
     source          1..3240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        542..576
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     CDS             603..1751
                     /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        1761..1852
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1853..2644
                     /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    2939..2955
                     /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 gggccgactc acccggcgcg gcgttgacag atgaggggca ggctcgattt cggccggcga
       61 cgtggagctg gccagcctcg caaatcggcg aaaacgcctg attttacgcg agtttcccac
      121 agatgatgtg gacaagcctg gggataagtg ccctgcggta ttgacacttg aggggcgcga
      181 ctactgacag atgaggggcg cgatccttga cacttgaggg gcagagtgct gacagatgag
      241 gggcgcacct attgacattt gaggggctgt ccacaggcag aaaatccagc atttgcaagg
      301 gtttccgccc gtttttcggc caccgctaac ctgtctttta acctgctttt aaaccaatat
      361 ttataaacct tgtttttaac cagggctgcg ccctgtgcgc gtgaccgcgc acgccgaagg
      421 ggggtgcccc cccttctcga accctcccgg cccgctaacg cgggcctccc atccccccag
      481 gggctgcgcc cctcggccgc gaacggcctc accccaaaaa tggaggtttc tcttctttca
      541 ctttacggct agctcagccc taggtattat gctagcagcc ccatacgata taaggagggt
      601 gaatgaatcg gacgtttgac cggaaggcat acaggcaaga actgatcgac gcggggtttt
      661 ccgccgagga tgccgaaacc atcgcaagcc gcaccgtcat gcgtgcgccc cgcgaaacct
      721 tccagtccgt cggctcgatg gtccagcaag ctacggccaa gatcgagcgc gacagcgtgc
      781 aactggctcc ccctgccctg cccgcgccat cggccgccgt ggagcgttcg cgtcgtctcg
      841 aacaggaggc ggcaggtttg gcgaagtcga tgaccatcga cacgcgagga actatgacga
      901 ccaagaagcg aaaaaccgcc ggcgaggacc tggcaaaaca ggtcagcgag gccaagcagg
      961 ccgcgttgct gaaacacacg aagcagcaga tcaaggaaat gcagctttcc ttgttcgata
     1021 ttgcgccgtg gccggacacg atgcgagcga tgccaaacga cacggcccgc tctgccctgt
     1081 tcaccacgcg caacaagaaa atcccgcgcg aggcgctgca aaacaaggtc attttccacg
     1141 tcaacaagga cgtgaagatc acctacaccg gcgtcgagct gcgggccgac gatgacgaac
     1201 tggtgtggca gcaggtgttg gagtacgcga agcgcacccc tatcggcgag ccgatcacct
     1261 tcacgttcta cgagctttgc caggacctgg gctggtcgat caatggccgg tattacacga
     1321 aggccgagga atgcctgtcg cgcctacagg cgacggcgat gggcttcacg tccgaccgcg
     1381 ttgggcacct ggaatcggtg tcgctgctgc accgcttccg cgtcctggac cgtggcaaga
     1441 aaacgtcccg ttgccaggtc ctgatcgacg aggaaatcgt cgtgctgttt gctggcgacc
     1501 actacacgaa attcatatgg gagaagtacc gcaagctgtc gccgacggcc cgacggatgt
     1561 tcgactattt cagctcgcac cgggagccgt acccgctcaa gctggaaacc ttccgcctca
     1621 tgtgcggatc ggattccacc cgcgtgaaga agtggcgcga gcaggtcggc gaagcctgcg
     1681 aagagttgcg aggcagcggc ctggtggaac acgcctgggt caatgatgac ctggtgcatt
     1741 gcaaacgcta gacggcctat ttggcctatt tttctaaata cattcaaata tgtatccgct
     1801 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgaggga
     1861 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     1921 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     1981 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     2041 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     2101 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     2161 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     2221 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     2281 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     2341 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     2401 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     2461 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     2521 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     2581 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     2641 ataaacgcca tgggcattaa aaaatgccct cttgggttat caagagggtc attatatttg
     2701 ctgaggagtg ccgttaatta agtcggatga atggcagaag cttaaaaaaa atccttagct
     2761 ttcgctaagg atctgcaggt cgacggatcc tccatcgcgt ggccgaaatt ggtggtgggg
     2821 gaaggattcg aaccttcgaa gtctgtgacg gcagatttag agttctgctc cctttggccg
     2881 ctcgggaacc ccaccggact tgatggtgcc gactaaaggg acccatcttg tctgctaatt
     2941 gttatccgct cacaattatc ttactgcgca gatacgccct cgtcaatccc ttaatagcaa
     3001 aatgcctttt gatcggcgag aaagtcagca ggccgcttag ttagccgtgc actgatcata
     3061 tcgtcaatta ttacctccac ggggagagcc tgagcaaact ggcctcaggc atttgagaag
     3121 cacaagggtc cctaagtctc ctcagcaaaa cgaaaggccc agtctttcga ctgagccttt
     3181 cgttttattt gaccggatgt cctcttactg acagatgagg ggcggacgtt gacacttgag
//
