LOCUS       sequence-266471-        3237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible expression of S-TAGA-Evo2 qtRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3237)
  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 3237)
  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-3
FEATURES             Location/Qualifiers
     source          1..3237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        442..476
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     CDS             503..1651
                     /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        1661..1752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1753..2544
                     /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    2836..2852
                     /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 attttacgcg agtttcccac agatgatgtg gacaagcctg gggataagtg ccctgcggta
       61 ttgacacttg aggggcgcga ctactgacag atgaggggcg cgatccttga cacttgaggg
      121 gcagagtgct gacagatgag gggcgcacct attgacattt gaggggctgt ccacaggcag
      181 aaaatccagc atttgcaagg gtttccgccc gtttttcggc caccgctaac ctgtctttta
      241 acctgctttt aaaccaatat ttataaacct tgtttttaac cagggctgcg ccctgtgcgc
      301 gtgaccgcgc acgccgaagg ggggtgcccc cccttctcga accctcccgg cccgctaacg
      361 cgggcctccc atccccccag gggctgcgcc cctcggccgc gaacggcctc accccaaaaa
      421 tggaggtttc tcttctttca ctttacggct agctcagccc taggtattat gctagcagcc
      481 ccatacgata taaggagggt gaatgaatcg gacgtttgac cggaaggcat acaggcaaga
      541 actgatcgac gcggggtttt ccgccgagga tgccgaaacc atcgcaagcc gcaccgtcat
      601 gcgtgcgccc cgcgaaacct tccagtccgt cggctcgatg gtccagcaag ctacggccaa
      661 gatcgagcgc gacagcgtgc aactggctcc ccctgccctg cccgcgccat cggccgccgt
      721 ggagcgttcg cgtcgtctcg aacaggaggc ggcaggtttg gcgaagtcga tgaccatcga
      781 cacgcgagga actatgacga ccaagaagcg aaaaaccgcc ggcgaggacc tggcaaaaca
      841 ggtcagcgag gccaagcagg ccgcgttgct gaaacacacg aagcagcaga tcaaggaaat
      901 gcagctttcc ttgttcgata ttgcgccgtg gccggacacg atgcgagcga tgccaaacga
      961 cacggcccgc tctgccctgt tcaccacgcg caacaagaaa atcccgcgcg aggcgctgca
     1021 aaacaaggtc attttccacg tcaacaagga cgtgaagatc acctacaccg gcgtcgagct
     1081 gcgggccgac gatgacgaac tggtgtggca gcaggtgttg gagtacgcga agcgcacccc
     1141 tatcggcgag ccgatcacct tcacgttcta cgagctttgc caggacctgg gctggtcgat
     1201 caatggccgg tattacacga aggccgagga atgcctgtcg cgcctacagg cgacggcgat
     1261 gggcttcacg tccgaccgcg ttgggcacct ggaatcggtg tcgctgctgc accgcttccg
     1321 cgtcctggac cgtggcaaga aaacgtcccg ttgccaggtc ctgatcgacg aggaaatcgt
     1381 cgtgctgttt gctggcgacc actacacgaa attcatatgg gagaagtacc gcaagctgtc
     1441 gccgacggcc cgacggatgt tcgactattt cagctcgcac cgggagccgt acccgctcaa
     1501 gctggaaacc ttccgcctca tgtgcggatc ggattccacc cgcgtgaaga agtggcgcga
     1561 gcaggtcggc gaagcctgcg aagagttgcg aggcagcggc ctggtggaac acgcctgggt
     1621 caatgatgac ctggtgcatt gcaaacgcta gacggcctat ttggcctatt tttctaaata
     1681 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1741 aaaaggaaga gtatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta
     1801 gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc
     1861 gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta
     1921 aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc
     1981 cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc
     2041 attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac
     2101 attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca
     2161 aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg
     2221 gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg
     2281 cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc
     2341 gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg
     2401 ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa
     2461 gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag
     2521 atcaccaagg tagtcggcaa ataaacgcca tgggcattaa aaaatgccct cttgggttat
     2581 caagagggtc attatatttg ctgaggagtg ccgttaatta agtcggatga atggcagaag
     2641 cttaaaaaaa atccttagct ttcgctaagg atctgcaggt cgacggatcc tgggaagtcg
     2701 ggagataagt cattgataaa gtggcggaga gagggggatt tgaacccccg gtagagttac
     2761 ccctactccg gtgttagaat accggtccgt tcagccgctc cggcatctct ccaagggacc
     2821 catcttgtct gctaattgtt atccgctcac aattatctta ctgcgcagat acgccctcgt
     2881 caatccctta atagcaaaat gccttttgat cggcgagaaa gtcagcaggc cgcttagtta
     2941 gccgtgcact gatcatatcg tcaattatta cctccacggg gagagcctga gcaaactggc
     3001 ctcaggcatt tgagaagcac aagggtccct aagtctcctc agcaaaacga aaggcccagt
     3061 ctttcgactg agcctttcgt tttatttgac cggatgtcct cttactgaca gatgaggggc
     3121 ggacgttgac acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc
     3181 tcgatttcgg ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctg
//
