LOCUS       sequence-266457-        3246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible expression of WT S tRNA scaffold tRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3246)
  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 3246)
  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: pED14xS-WT
FEATURES             Location/Qualifiers
     source          1..3246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        827..861
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     CDS             888..2036
                     /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        2046..2137
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2138..2929
                     /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    3230..3246
                     /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 ttatcttact gcgcagatac gccctcgtca atcccttaat agcaaaatgc cttttgatcg
       61 gcgagaaagt cagcaggccg cttagttagc cgtgcactga tcatatcgtc aattattacc
      121 tccacgggga gagcctgagc aaactggcct caggcatttg agaagcacaa gggtccctaa
      181 gtctcctcag caaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgaccg
      241 gatgtcctct tactgacaga tgaggggcgg acgttgacac ttgaggggcc gactcacccg
      301 gcgcggcgtt gacagatgag gggcaggctc gatttcggcc ggcgacgtgg agctggccag
      361 cctcgcaaat cggcgaaaac gcctgatttt acgcgagttt cccacagatg atgtggacaa
      421 gcctggggat aagtgccctg cggtattgac acttgagggg cgcgactact gacagatgag
      481 gggcgcgatc cttgacactt gaggggcaga gtgctgacag atgaggggcg cacctattga
      541 catttgaggg gctgtccaca ggcagaaaat ccagcatttg caagggtttc cgcccgtttt
      601 tcggccaccg ctaacctgtc ttttaacctg cttttaaacc aatatttata aaccttgttt
      661 ttaaccaggg ctgcgccctg tgcgcgtgac cgcgcacgcc gaaggggggt gccccccctt
      721 ctcgaaccct cccggcccgc taacgcgggc ctcccatccc cccaggggct gcgcccctcg
      781 gccgcgaacg gcctcacccc aaaaatggag gtttctcttc tttcacttta cggctagctc
      841 agccctaggt attatgctag cagccccata cgatataagg agggtgaatg aatcggacgt
      901 ttgaccggaa ggcatacagg caagaactga tcgacgcggg gttttccgcc gaggatgccg
      961 aaaccatcgc aagccgcacc gtcatgcgtg cgccccgcga aaccttccag tccgtcggct
     1021 cgatggtcca gcaagctacg gccaagatcg agcgcgacag cgtgcaactg gctccccctg
     1081 ccctgcccgc gccatcggcc gccgtggagc gttcgcgtcg tctcgaacag gaggcggcag
     1141 gtttggcgaa gtcgatgacc atcgacacgc gaggaactat gacgaccaag aagcgaaaaa
     1201 ccgccggcga ggacctggca aaacaggtca gcgaggccaa gcaggccgcg ttgctgaaac
     1261 acacgaagca gcagatcaag gaaatgcagc tttccttgtt cgatattgcg ccgtggccgg
     1321 acacgatgcg agcgatgcca aacgacacgg cccgctctgc cctgttcacc acgcgcaaca
     1381 agaaaatccc gcgcgaggcg ctgcaaaaca aggtcatttt ccacgtcaac aaggacgtga
     1441 agatcaccta caccggcgtc gagctgcggg ccgacgatga cgaactggtg tggcagcagg
     1501 tgttggagta cgcgaagcgc acccctatcg gcgagccgat caccttcacg ttctacgagc
     1561 tttgccagga cctgggctgg tcgatcaatg gccggtatta cacgaaggcc gaggaatgcc
     1621 tgtcgcgcct acaggcgacg gcgatgggct tcacgtccga ccgcgttggg cacctggaat
     1681 cggtgtcgct gctgcaccgc ttccgcgtcc tggaccgtgg caagaaaacg tcccgttgcc
     1741 aggtcctgat cgacgaggaa atcgtcgtgc tgtttgctgg cgaccactac acgaaattca
     1801 tatgggagaa gtaccgcaag ctgtcgccga cggcccgacg gatgttcgac tatttcagct
     1861 cgcaccggga gccgtacccg ctcaagctgg aaaccttccg cctcatgtgc ggatcggatt
     1921 ccacccgcgt gaagaagtgg cgcgagcagg tcggcgaagc ctgcgaagag ttgcgaggca
     1981 gcggcctggt ggaacacgcc tgggtcaatg atgacctggt gcattgcaaa cgctagacgg
     2041 cctatttggc ctatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2101 ctgataaatg cttcaataat attgaaaaag gaagagtatg agggaagcgg tgatcgccga
     2161 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     2221 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     2281 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     2341 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     2401 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     2461 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     2521 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     2581 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     2641 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     2701 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     2761 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     2821 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     2881 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataaa cgccatgggc
     2941 attaaaaaat gccctcttgg gttatcaaga gggtcattat atttgctgag gagtgccgtt
     3001 aattaagtcg gatgaatggc agaagcttaa aaaaaatcct tagctttcgc taaggatctg
     3061 caggtcgacg gatcctgaga taagtcattg ataaagtggc ggagagaggg ggatttgaac
     3121 ccccggtaga gttgccccta ctccggtttt cgagaccggt ccgttcagcc gctccggcat
     3181 ctctccgttc agatggttgc catgataagg gacccatctt gtctgctaat tgttatccgc
     3241 tcacaa
//
