LOCUS       sequence-112125-        5730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N2 inverter plasmid for two-strain system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5730)
  AUTHORS   Chen Y, Kim JK, Hirning AJ, Josic K, Bennett MR
  TITLE     SYNTHETIC BIOLOGY. Emergent genetic oscillations in a synthetic 
            microbial consortium.
  JOURNAL   Science. 2015 Aug 28;349(6251):986-9. doi: 10.1126/science.aaa3794.
  PUBMED    26315440
REFERENCE   2  (bases 1 to 5730)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2015-08-28- 28"; volume: "349"; 
            issue: "6251"; pages: "986-9"
COMMENT     Sequence Label: pC239
FEATURES             Location/Qualifiers
     source          1..5730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    873..891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    903..921
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(1075..1104)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    1305..1323
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1963..1981
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2182..2211
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    2365..2383
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2395..2413
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2497..3576
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2516..2535)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      3768..3795
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3898..4689)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATYDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(4870..4887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5004..5549)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5084..5103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5621..5640
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttattacgct gcaagggcgt aattttcgtc gttcgctgct atatattccg ggaacactct
       61 acaactcttt tcctgctcta tatcatgacc aaagaaaata attggtttct cttttttcac
      121 aacttctttt aaacgtttaa ttgaagataa agctaattct ggatcaaatc ctgcgaacgg
      181 cacttcatct tcaaaattct ctttcgtgta cgatgcatca atcgttaata aaactgaacc
      241 ggattgctcc gtctcaatga atagcgactg atggcctgga gaatgacctg gcgtatacaa
      301 taattgaaca cctggtacca cttcataatc cccttcaata attttgtagt tcaaatgcgg
      361 taatatacat tctttcatat attcttctct atgaagtgct gcctcatatt ccgttcgctg
      421 cacaataatt ggtgtatttg taaaagcacc gtttcctcct gcatgatcaa aatgtaagtg
      481 agaactaata atatataaaa ggtcgtccgg ctcataccct acacgcttta atatattcac
      541 gattctatct tcttcagtca ttttcggtaa gatctgtcct tcaacaaatg taccgttaaa
      601 aagcccttca ttattaactg cactttctgg catacctgtg tctactaaaa taggaccttc
      661 ttccgtctcc aaaagataac accacaccgg caagtttaat agtttccccg gtgttaacgc
      721 actgttaaca gacgaatgat ccaacatgca acgacctgct gggatgaaat aaagtttctt
      781 tactgtcata ctagtatttc tccttagcat gattaagatg tttcagtacg aaaattgctt
      841 tcattgttga tctccttttt aagtggcatg catctctatc actgataggg atgtgctcat
      901 tatctctatc actgataggg aagtaaattc cctcagatag gcccccccag atgaggggga
      961 tcagagcatg tcgcgcaaaa gtgtgcagcg gttttgcgat aacgacatgc gccacgcaca
     1021 agagttccgt agctgttcaa gtttgtgttt caactgttct cgtcgtttcc gcaacaagtc
     1081 ctcttcagaa atgagctttt gctcctctgc ttggacggac aggatgtatg ctgtggcttt
     1141 tttaaggata actaccttgg gggccttttc attgttttcc aactccggga tctggtcacg
     1201 cagggcaaaa aagctccgtt ttagctcgtt cctcctctgg cgctccaaga cgttgtgtgt
     1261 tcgcctcttg acattctcct cggtgtccga gggccctgtg tgaatctcta tcactgatag
     1321 ggatccacac agacgtctta acttgcttcc tagtgaaacc gttcaggaca tctacgggat
     1381 tgttgctaag aaagtcaacg agattctaca agcagacgca atcaatggga ccgataacga
     1441 agtagttacc gtgaccgatg agaacactgg tgaaatctct gagaaagtca agctgggcac
     1501 taaggcactg gctggtcaat ggctggctta cggtgttact cgcagtgtga ctaagcgttc
     1561 agtcatgacg ctggcttacg ggtccaaaga gttcggcttc cgtcaacaag tgctggaaga
     1621 taccattcag ccagctattg attccggcaa gggtctgatg ttcactcagc cgaatcaggc
     1681 tgctggatac atggctaagc tgatttggga atctgtgagc gtgacggtgg tagctgcggt
     1741 tgaagcaatg aactggctta agtctgctgc taagctgctg gctgctgagg tcaaagataa
     1801 gaagactgga gagattcttc gcaagcgttg cgctgtgcat tgggtaactc ctgatggttt
     1861 ccctgtgtgg caggaataca agaagcctat tcagacgcgc ttctcgagct tcccaacctt
     1921 accagagggc gccccagctg gcaattccga cgtctgtgtg gatccctatc agtgatagag
     1981 attcacacag ggccctcgga caccgaggag aatgtcaaga ggcgaacaca caacgtcttg
     2041 gagcgccaga ggaggaacga gctaaaacgg agcttttttg ccctgcgtga ccagatcccg
     2101 gagttggaaa acaatgaaaa ggcccccaag gtagttatcc ttaaaaaagc cacagcatac
     2161 atcctgtccg tccaagcaga ggagcaaaag ctcatttctg aagaggactt gttgcggaaa
     2221 cgacgagaac agttgaaaca caaacttgaa cagctacgga actcttgtgc gtaaggaaaa
     2281 gtaaggaaaa cgattccttc taacagaaat gtcctgagca atcacctatg aactgtcgag
     2341 ggggcctatc tgagggaatt tacttcccta tcagtgatag agatactgag cacatcccta
     2401 tcagtgatag agatgcatgc cacttaaaaa ggagatcaac aatgaaagca attttcgtac
     2461 tgaaacatct taatcatgct aaggagaaat actagtatga aaccagtaac gttatacgat
     2521 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     2581 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     2641 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     2701 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     2761 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     2821 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     2881 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     2941 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
     3001 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     3061 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     3121 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     3181 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     3241 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     3301 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc
     3361 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     3421 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     3481 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3541 ctggcacgac aggtttcccg actggaaagc gggcagacct ccgcagcgaa cgacgaaaat
     3601 tacgcccttg cagcgtaata aaagcttccc ctacgtgctg ctgaagttgc ccgcaacaga
     3661 gagtggaacc aaccggtgat accacgatac tatgactgag agtcaacgcc atgagcggcc
     3721 tcatttctta ttctgagtta caacagtccg caccgctgtc tcacactggc tcaccttcgg
     3781 gtgggccttt ctgcgtttat atactagaga gagaatataa aaagccagat tattaatccg
     3841 gcttttttat tatttcctag ctagcgatat ccgtcggctt gaacgaattg ttagacatta
     3901 tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc
     3961 tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat
     4021 gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg
     4081 cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg
     4141 ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc
     4201 aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc
     4261 aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcgt acgtggctgg
     4321 ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt
     4381 agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg
     4441 gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg
     4501 ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg
     4561 cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag
     4621 atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc
     4681 ttccctcatg atgtttaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     4741 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     4801 catagactgt accccaaaaa aacatgtcat aacaagaagc catgaaaacc gccggatata
     4861 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
     4921 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
     4981 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
     5041 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5101 ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt
     5161 ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt
     5221 tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc
     5281 cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc
     5341 cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg
     5401 aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg
     5461 tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag
     5521 agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt
     5581 tgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     5641 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     5701 tctgctcatg tttgacagct tatcgagctc
//
