LOCUS       sequence-261785-        5585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of 3WJ repressor index 19 trigger RNA via the 
            PluxB promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5585)
  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 5585)
  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: pYZ_PluxB_3WJrep_N19_trigger
FEATURES             Location/Qualifiers
     source          1..5585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(249..254,255..271,272..277))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(304..322)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        390..494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(495..563,564..1355)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(713..732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1526..2114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2015..2034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2268..2285
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2300..2439
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2365..2384)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2525..2547
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2541..2732)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4044..5126)
                     /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     5088..5107
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5127..5204)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5410..5429
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gggacctaac ataaacttgt taggtgcgta gatctgatta gtgtgagaaa aaagcccgca
       61 cctgacagtg cgggcttttt ttttcgagtc tacagactaa gcgcagaatt gtgtcgtcag
      121 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
      181 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
      241 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
      301 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
      361 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      421 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      481 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      541 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
      601 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
      661 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
      721 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
      781 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
      841 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
      901 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
      961 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1021 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     1081 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1141 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1201 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1261 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1321 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1381 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1441 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1501 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1561 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1621 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     1681 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1741 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1801 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1861 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1921 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1981 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2041 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2101 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2161 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2221 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2281 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2341 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2401 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     2461 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2521 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     2581 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     2641 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     2701 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     2761 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     2821 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     2881 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     2941 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     3001 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     3061 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     3121 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     3181 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     3241 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     3301 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     3361 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     3421 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     3481 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     3541 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     3601 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     3661 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     3721 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     3781 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     3841 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     3901 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     3961 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     4021 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4081 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     4141 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     4201 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     4261 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4321 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4381 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4441 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4501 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4561 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4621 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4681 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4741 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     4801 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     4861 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     4921 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     4981 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     5041 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     5101 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     5161 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     5221 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5281 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     5341 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5401 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5461 gccggccacg atgcgtccgg cgtagaggat cgagatctcg gaaatcaaac gcacagttcg
     5521 ttagtgtcgg acctgtagga tcgtacaggt ttacgcaaga aaatggtttg ttacagtcga
     5581 ataaa
//
