LOCUS       sequence-168532-        2285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Titration sponge for TetR and cI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2285)
  AUTHORS   Potvin-Trottier L, Lord ND, Vinnicombe G, Paulsson J
  TITLE     Synchronous long-term oscillations in a synthetic gene circuit.
  JOURNAL   Nature. 2016 Oct 12;538(7626):514-517. doi: 10.1038/nature19841.
  PUBMED    27732583
REFERENCE   2  (bases 1 to 2285)
  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"; journalName: 
            "Nature"; date: "2016-10-12- 12"; volume: "538"; issue: "7626"; 
            pages: "514-517"
COMMENT     Sequence Label: pLPT149
FEATURES             Location/Qualifiers
     source          1..2285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(6..24,25..30,31..47,48..53,54..79)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    6..24
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    31..49
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             89..100
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      227..313
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(478..1066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(558..577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1154..1248)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1279..2073)
                     /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(1390..1409)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2000..2019
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2252..2270
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
       61 tcagcaggac gcactgaccg aattcattaa agaggagaaa ggtaccgcag gcctgcagca
      121 aacgacgaaa actacgctgc atcagtttaa taagcttgat aacaccgtgc gtgttgacta
      181 ttttacctct ggcggtgaga tcccatggta cgcgtgctag aggcatcaaa taaaacgaaa
      241 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      301 gagtaggaca aatccgccgc cctagaccta gggcgttcgg ctgcggcgag cggtatcagc
      361 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      421 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      481 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      541 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      601 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      661 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
      721 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      781 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      841 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      901 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
      961 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1021 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1081 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1141 gactagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     1201 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctct
     1261 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     1321 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     1381 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     1441 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     1501 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
     1561 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     1621 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     1681 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     1741 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     1801 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     1861 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     1921 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     1981 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     2041 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     2101 atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     2161 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg acgtctaaga
     2221 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2281 tcacc
//
