LOCUS       sequence-168564-        2286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Titration sponge for TetR and lacI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2286)
  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 2286)
  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: pLPT148
FEATURES             Location/Qualifiers
     source          1..2286
                     /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)"
     protein_bind    160..176
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     protein_bind    183..199
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     terminator      228..314
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(479..1067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(559..578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1155..1249)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1280..2074)
                     /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(1391..1410)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2001..2020
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2253..2271
                     /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 tgtgagcgga taacaattga
      181 cattgtgagc ggataacaag atcccatggt acgcgtgcta gaggcatcaa ataaaacgaa
      241 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
      301 tgagtaggac aaatccgccg ccctagacct agggcgttcg gctgcggcga gcggtatcag
      361 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
      421 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      481 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      541 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      601 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      661 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      721 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      781 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      841 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      901 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
      961 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1021 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1081 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1141 tgactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac
     1201 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc
     1261 tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     1321 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     1381 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     1441 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     1501 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
     1561 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     1621 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     1681 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     1741 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     1801 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     1861 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     1921 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     1981 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     2041 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     2101 gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     2161 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc gacgtctaag
     2221 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     2281 ttcacc
//
