LOCUS       sequence-266459-        3274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible expression of W-TAGA-Evo1 qtRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3274)
  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 3274)
  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: pED14xW-3
FEATURES             Location/Qualifiers
     source          1..3274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        408..442
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     CDS             469..1617
                     /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        1627..1718
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1719..2510
                     /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"
     terminator      complement(2667..2710)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     protein_bind    2839..2855
                     /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 agcctgggga taagtgccct gcggtattga cacttgaggg gcgcgactac tgacagatga
       61 ggggcgcgat ccttgacact tgaggggcag agtgctgaca gatgaggggc gcacctattg
      121 acatttgagg ggctgtccac aggcagaaaa tccagcattt gcaagggttt ccgcccgttt
      181 ttcggccacc gctaacctgt cttttaacct gcttttaaac caatatttat aaaccttgtt
      241 tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc cgaagggggg tgccccccct
      301 tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc ccccaggggc tgcgcccctc
      361 ggccgcgaac ggcctcaccc caaaaatgga ggtttctctt ctttcacttt acggctagct
      421 cagccctagg tattatgcta gcagccccat acgatataag gagggtgaat gaatcggacg
      481 tttgaccgga aggcatacag gcaagaactg atcgacgcgg ggttttccgc cgaggatgcc
      541 gaaaccatcg caagccgcac cgtcatgcgt gcgccccgcg aaaccttcca gtccgtcggc
      601 tcgatggtcc agcaagctac ggccaagatc gagcgcgaca gcgtgcaact ggctccccct
      661 gccctgcccg cgccatcggc cgccgtggag cgttcgcgtc gtctcgaaca ggaggcggca
      721 ggtttggcga agtcgatgac catcgacacg cgaggaacta tgacgaccaa gaagcgaaaa
      781 accgccggcg aggacctggc aaaacaggtc agcgaggcca agcaggccgc gttgctgaaa
      841 cacacgaagc agcagatcaa ggaaatgcag ctttccttgt tcgatattgc gccgtggccg
      901 gacacgatgc gagcgatgcc aaacgacacg gcccgctctg ccctgttcac cacgcgcaac
      961 aagaaaatcc cgcgcgaggc gctgcaaaac aaggtcattt tccacgtcaa caaggacgtg
     1021 aagatcacct acaccggcgt cgagctgcgg gccgacgatg acgaactggt gtggcagcag
     1081 gtgttggagt acgcgaagcg cacccctatc ggcgagccga tcaccttcac gttctacgag
     1141 ctttgccagg acctgggctg gtcgatcaat ggccggtatt acacgaaggc cgaggaatgc
     1201 ctgtcgcgcc tacaggcgac ggcgatgggc ttcacgtccg accgcgttgg gcacctggaa
     1261 tcggtgtcgc tgctgcaccg cttccgcgtc ctggaccgtg gcaagaaaac gtcccgttgc
     1321 caggtcctga tcgacgagga aatcgtcgtg ctgtttgctg gcgaccacta cacgaaattc
     1381 atatgggaga agtaccgcaa gctgtcgccg acggcccgac ggatgttcga ctatttcagc
     1441 tcgcaccggg agccgtaccc gctcaagctg gaaaccttcc gcctcatgtg cggatcggat
     1501 tccacccgcg tgaagaagtg gcgcgagcag gtcggcgaag cctgcgaaga gttgcgaggc
     1561 agcggcctgg tggaacacgc ctgggtcaat gatgacctgg tgcattgcaa acgctagacg
     1621 gcctatttgg cctatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1681 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagggaagcg gtgatcgccg
     1741 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     1801 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     1861 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     1921 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     1981 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     2041 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     2101 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     2161 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     2221 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     2281 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     2341 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     2401 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     2461 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa acgccatggg
     2521 cattaaaaaa tgccctcttg ggttatcaag agggtcatta tatttgctga ggagtgccgt
     2581 taattaagtc ggatgaatgg cagaagctta aaaaaaatcc ttagctttcg ctaaggatct
     2641 acaggtcgac ggatcctgag agggttattt cagataaaaa aaatccttag ctttcgctaa
     2701 ggatgatttc tggcgggggc ggagagactc gaactcccaa cacccggtat tagaagaccg
     2761 gtgttctacc aattgaacta cgcccctaat tagggtggcg gaacggacgg gactcgaacc
     2821 cgcgatcttg tctgctaatt gttatccgct cacaattatc ttactgcgca gatacgccct
     2881 cgtcaatccc ttaatagcaa aatgcctttt gatcggcgag aaagtcagca ggccgcttag
     2941 ttagccgtgc actgatcata tcgtcaatta ttacctccac ggggagagcc tgagcaaact
     3001 ggcctcaggc atttgagaag cacaagggtc cctaagtctc ctcagcaaaa cgaaaggccc
     3061 agtctttcga ctgagccttt cgttttattt gaccggatgt cctcttactg acagatgagg
     3121 ggcggacgtt gacacttgag gggccgactc acccggcgcg gcgttgacag atgaggggca
     3181 ggctcgattt cggccggcga cgtggagctg gccagcctcg caaatcggcg aaaacgcctg
     3241 attttacgcg agtttcccac agatgatgtg gaca
//
