LOCUS       sequence-73804-e        4023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Positive feedback plasmid from oscillator without luxR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4023)
  AUTHORS   Prindle A, Samayoa P, Razinkov I, Danino T, Tsimring LS, Hasty J
  TITLE     A sensing array of radically coupled genetic 'biopixels'.
  JOURNAL   Nature. 2011 Dec 18;481(7379):39-44. doi: 10.1038/nature10722.
  PUBMED    22178928
REFERENCE   2  (bases 1 to 4023)
  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: "2011-12-18- 18"; volume: "481"; issue: "7379"; 
            pages: "39-44"
COMMENT     Sequence Label: pTD103luxI_sfGFP -R
FEATURES             Location/Qualifiers
     source          1..4023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             217..228
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      867..953
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             1181..1192
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1199..1201,1202..1912)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1199..1912
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1229..1257)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1852..1872
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1966..2052
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2217..2805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2297..2316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2893..2987)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3018..3812)
                     /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(3129..3148)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3739..3758
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3991..4009
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagagtc tataccgatg acgacgacct attgtttgtc gcaagttttg cgtgttatat
       61 atcattaaaa cggtaatgga ttgacatttg attctaataa attggatttt tgtcacacta
      121 ttgtatcgct gggaatacaa ttacttaaca taagcacctg taggatcgta caggtttacg
      181 caagaaaatg gtttgttata gtcgaatgaa ttcattaaag aggagaaagg taccatgact
      241 ataatgataa aaaaatcgga ttttttggca attccatcgg aggagtataa aggtattcta
      301 agtcttcgtt atcaagtgtt taagcaaaga cttgagtggg acttagttgt agaaaataac
      361 cttgaatcag atgagtatga taactcaaat gcagaatata tttatgcttg tgatgatact
      421 gaaaatgtaa gtggatgctg gcgtttatta cctacaacag gtgattatat gctgaaaagt
      481 gtttttcctg aattgcttgg tcaacagagt gctcccaaag atcctaatat agtcgaatta
      541 agtcgttttg ctgtaggtaa aaatagctca aagataaata actctgctag tgaaattaca
      601 atgaaactat ttgaagctat atataaacac gctgttagtc aaggtattac agaatatgta
      661 acagtaacat caacagcaat agagcgattt ttaaagcgta ttaaagttcc ttgtcatcgt
      721 attggagaca aagaaattca tgtattaggt gatactaaat cggttgtatt gtctatgcct
      781 attaatgaac agtttaaaaa agcagtctta aatgcagcga acgacgaaaa ttacgccctt
      841 gcagcgtaaa cgcgtgctag aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
      901 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc
      961 cctagaagac tgtcgctaca gatagttcac acctattgtt tgtcgcaagt tttgcgtgtt
     1021 atatatcatt aaaacggtaa tggattgaca tttgattcta ataaattgga tttttgtcac
     1081 actattgtat cgctgggaat acaattactt aacataagca cctgtaggat cgtacaggtt
     1141 tacgcaagaa aatggtttgt tatagtcgaa tgaattcatt aaagaggaga aaggtaccat
     1201 gagcaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
     1261 tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa ggtgatgcta caaacggaaa
     1321 actcaccctt aaatttattt gcactactgg aaaactacct gttccgtggc caacacttgt
     1381 cactactctg acctatggtg ttcaatgctt ttcccgttat ccggatcaca tgaaacggca
     1441 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa
     1501 agatgacggg acctacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa
     1561 tcgtatcgag ttaaagggta ttgattttaa agaagatgga aacattcttg gacacaaact
     1621 cgagtacaac tttaactcac acaatgtata catcacggca gacaaacaaa agaatggaat
     1681 caaagctaac ttcaaaattc gccacaacgt tgaagatggt tccgttcaac tagcagacca
     1741 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     1801 gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct
     1861 tgagtttgta actgctgctg ggattacaca tggcatggat gagctctaca aagcagcgaa
     1921 cgacgaaaat tacgcccttg cagcgtgaac gcgtgctaga ggcatcaaat aaaacgaaag
     1981 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     2041 agtaggacaa atccgccgcc ctagacctag ggcgttcggc tgcggcgagc ggtatcagct
     2101 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2161 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2221 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2281 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2341 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2401 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2461 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2521 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2581 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2641 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2701 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2761 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2821 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2881 actagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     2941 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctctc
     3001 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     3061 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     3121 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     3181 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     3241 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
     3301 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     3361 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     3421 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     3481 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     3541 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     3601 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     3661 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     3721 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     3781 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     3841 tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
     3901 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccga cgtctaagaa
     3961 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     4021 cac
//
