LOCUS       sequence-350201-        3956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Toehold switch, with an unpaired toehold region at the 5′-end that 
            interacts with the trigger RNA (sTR060, Addgene plasmid # 177370) to
            unfold the switch..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3956)
  AUTHORS   Batista AC, Levrier A, Soudier P, Voyvodic PL, Achmedov T, 
            Reif-Trauttmansdorff T, DeVisch A, Cohen-Gonsaud M, Faulon JL, 
            Beisel CL, Bonnet J, Kushwaha M
  TITLE     Differentially Optimized Cell-Free Buffer Enables Robust Expression 
            from Unprotected Linear DNA in Exonuclease-Deficient Extracts.
  JOURNAL   ACS Synth Biol. 2022 Jan 16. doi: 10.1021/acssynbio.1c00448.
  PUBMED    35034449
REFERENCE   2  (bases 1 to 3956)
  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: "ACS Synth 
            Biol. 2022 Jan 16. doi: 10.1021/acssynbio.1c00448."
COMMENT     Sequence Label: sTR056
FEATURES             Location/Qualifiers
     source          1..3956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        97..115
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             210..923
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1065..1159
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1395..2186
                     /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"
     oriT            2332..2440
                     /note="incP origin of transfer"
     CDS             complement(2631..3581)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3629..3851
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(join(3952..3956,1..82))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
       61 actgagcctt tcgttttatt tgatgccttt aattaataat acgactcact ataggagcag
      121 ggtcttatct tatctatctc gtttatccct gcatacagaa acagaggaga tatgcaatga
      181 taaacgagaa cctggcggca gcgcaaaaga tgcgtaaagg cgaagaactg tttaccggtg
      241 tggttccgat tctggtggaa ctggacggcg atgttaatgg tcataaattc agtgttcgcg
      301 gcgaaggtga aggcgatgcg acgaacggca aactgaccct gaaatttatc tgcaccacgg
      361 gtaaactgcc ggtcccgtgg ccgacgctgg tgaccacgct gacctatggc gttcaatgtt
      421 ttgcgcgtta cccggatcac atgaaacagc acgacttttt caaatcggcc atgccggaag
      481 gctatgtgca ggaacgtacg attagcttta aagacgatgg tacgtataaa acccgcgcgg
      541 aagtgaaatt cgaaggcgat accctggtta accgtatcga actgaaaggt atcgatttca
      601 aagaagacgg caatattctg ggtcataaac tggaatataa cttcaattcc cacaacgtgt
      661 acatcaccgc ggataaacag aaaaacggca ttaaagccaa tttcaaaatc cgccataatg
      721 tggaagatgg tagcgttcag ctggccgacc actatcagca aaacacgccg attggtgatg
      781 gcccggtcct gctgccggac aatcactacc tgagtaccca gtccgtgctg tcaaaagatc
      841 cgaacgaaaa acgtgaccac atggtcctgc tggaatttgt gacggctgcg ggtatcaccc
      901 acggcatgga cgaactgtat aaaaggcctg cagcaaacga cgaaaactac gctgcatcag
      961 tttaataata ctcgaacccc tagcccgctc ttatcgggcg gctaggggtt ttttgtccta
     1021 gggcggccgc gtcgtgactg ggaaaaccct ggcgactagt cttggactcc tgttgataga
     1081 tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc
     1141 gttttttatt ggtgagaatc caggggtccc caataattac gatttacgta tttaaatgaa
     1201 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     1261 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     1321 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     1381 caaagttaaa catcatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
     1441 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     1501 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     1561 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     1621 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     1681 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     1741 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     1801 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     1861 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     1921 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     1981 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     2041 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
     2101 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
     2161 agatcaccaa ggtagtcggc aaataagaca attgtccttt tccgctgcat aaccctgctt
     2221 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     2281 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat
     2341 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     2401 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg taggccggcc tggtgaacag
     2461 ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt
     2521 catctgtgca tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt
     2581 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
     2641 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     2701 gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     2761 agctgaattt ttgctgttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag
     2821 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     2881 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     2941 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     3001 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     3061 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     3121 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     3181 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     3241 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaag
     3301 atcttaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     3361 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     3421 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     3481 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     3541 atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg
     3601 tgattttaat cactatacca attgagatgg gctagtcaat gataattact agtccttttc
     3661 ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct
     3721 gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg
     3781 gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct
     3841 gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg
     3901 tttgctcctc tacaaaacag acctggcgcg cccagctgtc tagggcggcg gatttg
//
