LOCUS       sequence-168534-        2269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Titration sponge for TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2269)
  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 2269)
  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: pLPT41
FEATURES             Location/Qualifiers
     source          1..2269
                     /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      211..297
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(462..1050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(542..561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1138..1232)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1263..2057)
                     /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(1374..1393)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1984..2003
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2236..2254
                     /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 atcgaattcc tgcagcccgg
      181 gggatcccat ggtacgcgtg ctagaggcat caaataaaac gaaaggctca gtcgaaagac
      241 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
      301 ccgccctaga cctagggcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      361 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      421 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      481 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      541 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      601 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      661 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      721 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      781 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      841 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      901 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      961 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1021 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1081 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgactag tgcttggatt
     1141 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     1201 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctctcgaacc ccagagtccc
     1261 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     1321 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     1381 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     1441 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     1501 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     1561 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     1621 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     1681 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     1741 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     1801 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     1861 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     1921 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     1981 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     2041 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
     2101 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     2161 accttaccag agggcgcccc agctggcaat tccgacgtct aagaaaccat tattatcatg
     2221 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcacc
//
