LOCUS       sequence-266454-        3239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible expression of S-TAGA-Evo3 qtRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3239)
  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 3239)
  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-4
FEATURES             Location/Qualifiers
     source          1..3239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        332..366
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     CDS             393..1541
                     /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        1551..1642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1643..2434
                     /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    2728..2744
                     /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 gcagagtgct gacagatgag gggcgcacct attgacattt gaggggctgt
       61 ccacaggcag aaaatccagc atttgcaagg gtttccgccc gtttttcggc caccgctaac
      121 ctgtctttta acctgctttt aaaccaatat ttataaacct tgtttttaac cagggctgcg
      181 ccctgtgcgc gtgaccgcgc acgccgaagg ggggtgcccc cccttctcga accctcccgg
      241 cccgctaacg cgggcctccc atccccccag gggctgcgcc cctcggccgc gaacggcctc
      301 accccaaaaa tggaggtttc tcttctttca ctttacggct agctcagccc taggtattat
      361 gctagcagcc ccatacgata taaggagggt gaatgaatcg gacgtttgac cggaaggcat
      421 acaggcaaga actgatcgac gcggggtttt ccgccgagga tgccgaaacc atcgcaagcc
      481 gcaccgtcat gcgtgcgccc cgcgaaacct tccagtccgt cggctcgatg gtccagcaag
      541 ctacggccaa gatcgagcgc gacagcgtgc aactggctcc ccctgccctg cccgcgccat
      601 cggccgccgt ggagcgttcg cgtcgtctcg aacaggaggc ggcaggtttg gcgaagtcga
      661 tgaccatcga cacgcgagga actatgacga ccaagaagcg aaaaaccgcc ggcgaggacc
      721 tggcaaaaca ggtcagcgag gccaagcagg ccgcgttgct gaaacacacg aagcagcaga
      781 tcaaggaaat gcagctttcc ttgttcgata ttgcgccgtg gccggacacg atgcgagcga
      841 tgccaaacga cacggcccgc tctgccctgt tcaccacgcg caacaagaaa atcccgcgcg
      901 aggcgctgca aaacaaggtc attttccacg tcaacaagga cgtgaagatc acctacaccg
      961 gcgtcgagct gcgggccgac gatgacgaac tggtgtggca gcaggtgttg gagtacgcga
     1021 agcgcacccc tatcggcgag ccgatcacct tcacgttcta cgagctttgc caggacctgg
     1081 gctggtcgat caatggccgg tattacacga aggccgagga atgcctgtcg cgcctacagg
     1141 cgacggcgat gggcttcacg tccgaccgcg ttgggcacct ggaatcggtg tcgctgctgc
     1201 accgcttccg cgtcctggac cgtggcaaga aaacgtcccg ttgccaggtc ctgatcgacg
     1261 aggaaatcgt cgtgctgttt gctggcgacc actacacgaa attcatatgg gagaagtacc
     1321 gcaagctgtc gccgacggcc cgacggatgt tcgactattt cagctcgcac cgggagccgt
     1381 acccgctcaa gctggaaacc ttccgcctca tgtgcggatc ggattccacc cgcgtgaaga
     1441 agtggcgcga gcaggtcggc gaagcctgcg aagagttgcg aggcagcggc ctggtggaac
     1501 acgcctgggt caatgatgac ctggtgcatt gcaaacgcta gacggcctat ttggcctatt
     1561 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1621 ataatattga aaaaggaaga gtatgaggga agcggtgatc gccgaagtat cgactcaact
     1681 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     1741 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     1801 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     1861 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     1921 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     1981 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     2041 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     2101 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     2161 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     2221 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     2281 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     2341 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     2401 gaaaggcgag atcaccaagg tagtcggcaa ataaacgcca tgggcattaa aaaatgccct
     2461 cttgggttat caagagggtc attatatttg ctgaggagtg ccgttaatta agtcggatga
     2521 atggcagaag cttaaaaaaa aaatccttag ctttcgctaa ggatctgcag gtcgacggat
     2581 cctgggaagt cgggagataa gtcattgata aagtggcgga gagaggggga tttgaacccc
     2641 cggtagagtt gtccctactc cggggttaga atcccggtcc gttcagccgc tccggcatct
     2701 ctccaaggga cccatcttgt ctgctaattg ttatccgctc acaattatct tactgcgcag
     2761 atacgccctc gtcaatccct taatagcaaa atgccttttg atcggcgaga aagtcagcag
     2821 gccgcttagt tagccgtgca ctgatcatat cgtcaattat tacctccacg gggagagcct
     2881 gagcaaactg gcctcaggca tttgagaagc acaagggtcc ctaagtctcc tcagcaaaac
     2941 gaaaggccca gtctttcgac tgagcctttc gttttatttg accggatgtc ctcttactga
     3001 cagatgaggg gcggacgttg acacttgagg ggccgactca cccggcgcgg cgttgacaga
     3061 tgaggggcag gctcgatttc ggccggcgac gtggagctgg ccagcctcgc aaatcggcga
     3121 aaacgcctga ttttacgcga gtttcccaca gatgatgtgg acaagcctgg ggataagtgc
     3181 cctgcggtat tgacacttga ggggcgcgac tactgacaga tgaggggcgc gatccttga
//
