LOCUS       sequence-259953-        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"
     promoter        complement(6..97)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(107..1255)
                     /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        complement(1282..1316)
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     protein_bind    2143..2159
                     /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)."
     CDS             complement(join(2448..3234,1..5))
                     /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 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
       61 ggatacatat ttgaatgtat ttagaaaaat aggccaaata ggccgtctag cgtttgcaat
      121 gcaccaggtc atcattgacc caggcgtgtt ccaccaggcc gctgcctcgc aactcttcgc
      181 aggcttcgcc gacctgctcg cgccacttct tcacgcgggt ggaatccgat ccgcacatga
      241 ggcggaaggt ttccagcttg agcgggtacg gctcccggtg cgagctgaaa tagtcgaaca
      301 tccgtcgggc cgtcggcgac agcttgcggt acttctccca tatgaatttc gtgtagtggt
      361 cgccagcaaa cagcacgacg atttcctcgt cgatcaggac ctggcaacgg gacgttttct
      421 tgccacggtc caggacgcgg aagcggtgca gcagcgacac cgattccagg tgcccaacgc
      481 ggtcggacgt gaagcccatc gccgtcgcct gtaggcgcga caggcattcc tcggccttcg
      541 tgtaataccg gccattgatc gaccagccca ggtcctggca aagctcgtag aacgtgaagg
      601 tgatcggctc gccgataggg gtgcgcttcg cgtactccaa cacctgctgc cacaccagtt
      661 cgtcatcgtc ggcccgcagc tcgacgccgg tgtaggtgat cttcacgtcc ttgttgacgt
      721 ggaaaatgac cttgttttgc agcgcctcgc gcgggatttt cttgttgcgc gtggtgaaca
      781 gggcagagcg ggccgtgtcg tttggcatcg ctcgcatcgt gtccggccac ggcgcaatat
      841 cgaacaagga aagctgcatt tccttgatct gctgcttcgt gtgtttcagc aacgcggcct
      901 gcttggcctc gctgacctgt tttgccaggt cctcgccggc ggtttttcgc ttcttggtcg
      961 tcatagttcc tcgcgtgtcg atggtcatcg acttcgccaa acctgccgcc tcctgttcga
     1021 gacgacgcga acgctccacg gcggccgatg gcgcgggcag ggcaggggga gccagttgca
     1081 cgctgtcgcg ctcgatcttg gccgtagctt gctggaccat cgagccgacg gactggaagg
     1141 tttcgcgggg cgcacgcatg acggtgcggc ttgcgatggt ttcggcatcc tcggcggaaa
     1201 accccgcgtc gatcagttct tgcctgtatg ccttccggtc aaacgtccga ttcattcacc
     1261 ctccttatat cgtatggggc tgctagcata atacctaggg ctgagctagc cgtaaagtga
     1321 aagaagagaa acctccattt ttggggtgag gccgttcgcg gccgaggggc gcagcccctg
     1381 gggggatggg aggcccgcgt tagcgggccg ggagggttcg agaagggggg gcacccccct
     1441 tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg tttataaata
     1501 ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg cggaaaccct
     1561 tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc
     1621 atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg
     1681 cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac atcatctgtg
     1741 ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag ctccacgtcg
     1801 ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag tcggcccctc
     1861 aagtgtcaac gtccgcccct catctgtcag taagaggaca tccggtcaaa taaaacgaaa
     1921 ggctcagtcg aaagactggg cctttcgttt tgctgaggag acttagggac ccttgtgctt
     1981 ctcaaatgcc tgaggccagt ttgctcaggc tctccccgtg gaggtaataa ttgacgatat
     2041 gatcagtgca cggctaacta agcggcctgc tgactttctc gccgatcaaa aggcattttg
     2101 ctattaaggg attgacgagg gcgtatctgc gcagtaagat aattgtgagc ggataacaat
     2161 tagcagacaa gatgggtccc ttgtagaata agttttcccg atgcatccgt agttcagctg
     2221 gatagagcac tcggcttcta aaccgatcgg tcggaggttc gaatcctccc ggatgcacca
     2281 tattctccgt aaccttcagc aggatccgtc gacctgcaga tccttagcga aagctaagga
     2341 ttttttttaa gcttctgcca ttcatccgac ttaattaacg gcactcctca gcaaatataa
     2401 tgaccctctt gataacccaa gagggcattt tttaatgccc atggcgttta tttgccgact
     2461 accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag
     2521 gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga
     2581 tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg
     2641 ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca
     2701 gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc
     2761 tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt
     2821 tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata
     2881 cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa
     2941 cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg
     3001 ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt
     3061 tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg
     3121 ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg
     3181 atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttcc
//
