LOCUS       sequence-168539-        2490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Titration sponge for TetR, cI and lacI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2490)
  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 2490)
  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: pLPT145
FEATURES             Location/Qualifiers
     source          1..2490
                     /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    247..263
                     /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    388..404
                     /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      433..519
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(683..1271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(763..782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1359..1453)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1484..2278)
                     /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(1595..1614)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2205..2224
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
       61 tcagcaggac gcactgaccg aattcattaa agaggagaaa ggtaccgcag gcctgcagca
      121 aacgacgaaa actacgctgc atcagtttaa taagcttgag caaccattat caccgccaga
      181 ggtaaaatag tcaacacgca cggtgttaga tatttatcca aggaacgcca gccgccacga
      241 cgtttgttgt gagcggataa caagtggaat gtcttttgtg acccgaaaag tgccacctga
      301 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
      361 ctttcgtctt cacctcgaga attgacattg tgagcggata acaagatccc atggtacgcg
      421 tgctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      481 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctaggcg
      541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
      601 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      661 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
      721 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      781 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      841 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      901 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1021 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1081 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     1141 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1201 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1261 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1321 actcacgtta agggattttg gtcatgacta gtgcttggat tctcaccaat aaaaaacgcc
     1381 cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct
     1441 atcaacagga gtccaagcga gctctcgaac cccagagtcc cgctcagaag aactcgtcaa
     1501 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     1561 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     1621 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     1681 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     1741 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     1801 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     1861 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     1921 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     1981 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     2041 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca
     2101 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     2161 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     2221 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     2281 gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg
     2341 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
     2401 cagctggcaa ttccgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     2461 ggcgtatcac gaggcccttt cgtcttcacc
//
