LOCUS       sequence-261807-        3411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP-driven expression of first-generation toehold repressor 
            index 9 trigger RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3411)
  AUTHORS   Kim J, Zhou Y, Carlson PD, Teichmann M, Chaudhary S, Simmel FC, 
            Silver PA, Collins JJ, Lucks JB, Yin P, Green AA
  TITLE     De novo-designed translation-repressing riboregulators for 
            multi-input cellular logic.
  JOURNAL   Nat Chem Biol. 2019 Dec;15(12):1173-1182. doi: 
            10.1038/s41589-019-0388-1. Epub 2019 Nov 4.
  PUBMED    31686032
REFERENCE   2  (bases 1 to 3411)
  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"; doi: 
            "10.1038/s41589-019-0388-1"; journalName: "Nat Chem Biol"; date: 
            "2019-12"; volume: "15"; issue: "12"; pages: "1173-1182"
COMMENT     Sequence Label: pAG_ToeRep_N09_trigger
FEATURES             Location/Qualifiers
     source          1..3411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      82..128
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(299..1090)
                     /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"
     promoter        complement(1091..1182)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1230..1968)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(2178..3260)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3222..3241
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3261..3338)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3396..3411,1..4)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3396..3411,1..3)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agggtcaagg cattagccct aatgccttga atacttacaa tcatcgcctt cattcaactt
       61 tatattcttc ttcactttac ttagcataac cccttggggc ctctaaacgg gtcttgaggg
      121 gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
      181 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
      241 cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt gttagacatt
      301 atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat
      361 ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta
      421 tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg
      481 gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc
      541 gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct
      601 caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg
      661 caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg
      721 gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct
      781 tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc
      841 ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc
      901 gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg
      961 gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga
     1021 gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg
     1081 cttccctcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1141 tgagcggata catatttgaa tgtatttaga aaaataaaca aatagctagc tcactcggtc
     1201 gctacgctcc gggcgtgaga ctgcggcggg cgctgcggac acatacaaag ttacccacag
     1261 attccgtgga taagcagggg actaacatgt gaggcaaaac agcagggccg cgccggtggc
     1321 gtttttccat aggctccgcc ctcctgccag agttcacata aacagacgct tttccggtgc
     1381 atctgtggga gccgtgaggc tcaaccatga atctgacagt acgggcgaaa cccgacagga
     1441 cttaaagatc cccaccgttt ccggcgggtc gctccctctt gcgctctcct gttccgaccc
     1501 tgccgtttac cggatacctg ttccgccttt ctcccttacg ggaagtgtgg cgctttctca
     1561 tagctcacac actggtatct cggctcggtg taggtcgttc gctccaagct gggctgtaag
     1621 caagaactcc ccgttcagcc cgactgctgc gccttatccg gtaactgttc acttgagtcc
     1681 aacccggaaa agcacggtaa aacgccactg gcagcagcca ttggtaactg ggagttcgca
     1741 gaggatttgt ttagctaaac acgcggttgc tcttgaagtg tgcgccaaag tccggctaca
     1801 ctggaaggac agatttggtt gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt
     1861 ccccaactga cttaaccttc gatcaaacca cctccccagg tggttttttc gtttacaggg
     1921 caaaagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctactgaa
     1981 ccgctctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     2041 cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg
     2101 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     2161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2221 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     2281 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     2341 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     2401 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     2461 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     2521 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     2581 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     2641 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     2701 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     2761 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     2821 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     2881 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     2941 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     3001 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     3061 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     3121 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     3181 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     3241 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     3301 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     3361 atgaacgtgt acgggctatc tggctttcgt tgcgctaata cgactcacta t
//
