LOCUS       sequence-459807-        2279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MicF RNA toehold switch with NanoLuc complementation peptide 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2279)
  AUTHORS   Diaz KJ, Jarquin J, Petrosyan A, Takahashi MK
  TITLE     Improved RNA toehold switch sensitivity using the NanoLuc 
            complementation reporter.
  JOURNAL   MicroPubl Biol. 2025 Jan 2;2025:10.17912/micropub.biology.001334. 
            doi: 10.17912/micropub.biology.001334. eCollection 2025.
  PUBMED    39839716
REFERENCE   2  (bases 1 to 2279)
  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: "MicroPubl 
            Biol."; date: "2025-01-2"; volume: 
            "2025:10.17912/micropub.biology.001334. doi"; pages: " 
            10.17912/micropub.biology.001334. eCollection 2025"
COMMENT     Sequence Label: MKT074
FEATURES             Location/Qualifiers
     source          1..2279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..197
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        224..258
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     terminator      433..519
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(684..1272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(764..783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1360..1454)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1485..2279)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1596..1615)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2206..2225
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct
       61 tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc
      121 cccagctggc aattccgacg tctaagaaac cattattatc atgacattaa cctataaaaa
      181 taggcgtatc acgaggccct ttcgtcttca cctcgaggta cccttgacag ctagctcagt
      241 cctaggtata atactagtgt gaatgacggt aataaataaa gttaatgatg atagcggact
      301 ttagaacaga ggagataaag atggctatca tcatgggagg ttctggaggt gtcagcggat
      361 ggcgactctt caagaagatc agctaataag gtgcttaata aggatcctgt cgatcgcgcg
      421 tgctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      481 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagggc
      541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1081 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     1141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1321 aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa taaaaaacgc
     1381 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     1441 tatcaacagg agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca
     1501 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     1561 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     1621 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     1681 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     1741 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     1801 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     1861 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     1921 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     1981 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     2041 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     2101 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     2161 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     2221 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcat
//
