LOCUS       sequence-266470-        3234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible expression of R-TAGA-Evo2 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-4
FEATURES             Location/Qualifiers
     source          1..3234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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)."
     promoter        844..878
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     CDS             905..2053
                     /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        2063..2154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2155..2946
                     /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"
ORIGIN
        1 ttgttatccg ctcacaatta tcttactgcg cagatacgcc ctcgtcaatc ccttaatagc
       61 aaaatgcctt ttgatcggcg agaaagtcag caggccgctt agttagccgt gcactgatca
      121 tatcgtcaat tattacctcc acggggagag cctgagcaaa ctggcctcag gcatttgaga
      181 agcacaaggg tccctaagtc tcctcagcaa aacgaaaggc ccagtctttc gactgagcct
      241 ttcgttttat ttgaccggat gtcctcttac tgacagatga ggggcggacg ttgacacttg
      301 aggggccgac tcacccggcg cggcgttgac agatgagggg caggctcgat ttcggccggc
      361 gacgtggagc tggccagcct cgcaaatcgg cgaaaacgcc tgattttacg cgagtttccc
      421 acagatgatg tggacaagcc tggggataag tgccctgcgg tattgacact tgaggggcgc
      481 gactactgac agatgagggg cgcgatcctt gacacttgag gggcagagtg ctgacagatg
      541 aggggcgcac ctattgacat ttgaggggct gtccacaggc agaaaatcca gcatttgcaa
      601 gggtttccgc ccgtttttcg gccaccgcta acctgtcttt taacctgctt ttaaaccaat
      661 atttataaac cttgttttta accagggctg cgccctgtgc gcgtgaccgc gcacgccgaa
      721 ggggggtgcc cccccttctc gaaccctccc ggcccgctaa cgcgggcctc ccatcccccc
      781 aggggctgcg cccctcggcc gcgaacggcc tcaccccaaa aatggaggtt tctcttcttt
      841 cactttacgg ctagctcagc cctaggtatt atgctagcag ccccatacga tataaggagg
      901 gtgaatgaat cggacgtttg accggaaggc atacaggcaa gaactgatcg acgcggggtt
      961 ttccgccgag gatgccgaaa ccatcgcaag ccgcaccgtc atgcgtgcgc cccgcgaaac
     1021 cttccagtcc gtcggctcga tggtccagca agctacggcc aagatcgagc gcgacagcgt
     1081 gcaactggct ccccctgccc tgcccgcgcc atcggccgcc gtggagcgtt cgcgtcgtct
     1141 cgaacaggag gcggcaggtt tggcgaagtc gatgaccatc gacacgcgag gaactatgac
     1201 gaccaagaag cgaaaaaccg ccggcgagga cctggcaaaa caggtcagcg aggccaagca
     1261 ggccgcgttg ctgaaacaca cgaagcagca gatcaaggaa atgcagcttt ccttgttcga
     1321 tattgcgccg tggccggaca cgatgcgagc gatgccaaac gacacggccc gctctgccct
     1381 gttcaccacg cgcaacaaga aaatcccgcg cgaggcgctg caaaacaagg tcattttcca
     1441 cgtcaacaag gacgtgaaga tcacctacac cggcgtcgag ctgcgggccg acgatgacga
     1501 actggtgtgg cagcaggtgt tggagtacgc gaagcgcacc cctatcggcg agccgatcac
     1561 cttcacgttc tacgagcttt gccaggacct gggctggtcg atcaatggcc ggtattacac
     1621 gaaggccgag gaatgcctgt cgcgcctaca ggcgacggcg atgggcttca cgtccgaccg
     1681 cgttgggcac ctggaatcgg tgtcgctgct gcaccgcttc cgcgtcctgg accgtggcaa
     1741 gaaaacgtcc cgttgccagg tcctgatcga cgaggaaatc gtcgtgctgt ttgctggcga
     1801 ccactacacg aaattcatat gggagaagta ccgcaagctg tcgccgacgg cccgacggat
     1861 gttcgactat ttcagctcgc accgggagcc gtacccgctc aagctggaaa ccttccgcct
     1921 catgtgcgga tcggattcca cccgcgtgaa gaagtggcgc gagcaggtcg gcgaagcctg
     1981 cgaagagttg cgaggcagcg gcctggtgga acacgcctgg gtcaatgatg acctggtgca
     2041 ttgcaaacgc tagacggcct atttggccta tttttctaaa tacattcaaa tatgtatccg
     2101 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagg
     2161 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
     2221 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     2281 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     2341 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     2401 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     2461 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     2521 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     2581 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     2641 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     2701 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     2761 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     2821 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     2881 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     2941 aaataaacgc catgggcatt aaaaaatgcc ctcttgggtt atcaagaggg tcattatatt
     3001 tgctgaggag tgccgttaat taagtcggat gaatggcaga agcttaaaaa aaatccttag
     3061 ctttcgctaa ggatctgcag gtcgacggat cctgctgaag gttacggaga atatggtgca
     3121 tccgggagga ttcgaacctc cgaccgatcg gtttagaagc cgagtgctct atccagctga
     3181 actacggatg catcgggaaa acttattcta caagggaccc atcttgtctg ctaa
//
