LOCUS       sequence-483545-        5542 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  Positive feedback plasmid from oscillator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5542)
  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 5542)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 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
FEATURES             Location/Qualifiers
     source          1..5542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             968..979
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(986..988,989..1699)
                     /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             986..1699
                     /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(1016..1044)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1639..1659
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1753..1839
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2835..2846
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3485..3571
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3736..4324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3816..3835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4412..4506)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4537..5331)
                     /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(4648..4667)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5258..5277
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5510..5528
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagttaa tttttaaagt atgggcaatc aattgctcct gttaaaattg ctttagaaat
       61 actttggcag cggtttgttg tattgagttt catttgcgca ttggttaaat ggaaagtgac
      121 agtacgctca ctgcagccta atatttttga aatatcccaa gagctttttc cttcgcatgc
      181 ccacgctaaa cattcttttt ctcttttggt taaatcgttg tttgatttat tatttgctat
      241 atttattttt cgataattat caactagaga aggaacaatt aatggtatgt tcatacacgc
      301 atgtaaaaat aaactatcta tatagttgtc tttttctgaa tgtgcaaaac taagcattcc
      361 gaagccattg ttagccgtat gaatagggaa actaaaccca gtgataagac ctgatgtttt
      421 cgcttcttta attacatttg gagatttttt atttacagca ttgttttcaa atatattcca
      481 attaattggt gaatgattgg agttagaata atctactata ggatcatatt ttattaaatt
      541 agcgtcatca taatattgcc tccatttttt agggtaatta tctagaattg aaatatcaga
      601 tttaaccata gaatgaggat aaatgatcgc gagtaaataa tattcacaat gtaccatttt
      661 agtcatatca gataagcatt gattaatatc attattgctt ctacaagctt taattttatt
      721 aattattctg tatgtgtcgt cggcatttat gtttttcata cccatctctt tatccttacc
      781 tattgtttgt cgcaagtttt gcgtgttata tatcattaaa acggtaatgg attgacattt
      841 gattctaata aattggattt ttgtcacact attgtatcgc tgggaataca attacttaac
      901 ataagcacct gtaggatcgt acaggtttac gcaagaaaat ggtttgttat agtcgaatga
      961 attcattaaa gaggagaaag gtaccatgag caaaggagaa gaacttttca ctggagttgt
     1021 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tccgtggaga
     1081 gggtgaaggt gatgctacaa acggaaaact cacccttaaa tttatttgca ctactggaaa
     1141 actacctgtt ccgtggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc
     1201 ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta
     1261 tgtacaggaa cgcactatat ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt
     1321 caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga
     1381 agatggaaac attcttggac acaaactcga gtacaacttt aactcacaca atgtatacat
     1441 cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacgttga
     1501 agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     1561 tgtcctttta ccagacaacc attacctgtc gacacaatct gtcctttcga aagatcccaa
     1621 cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg
     1681 catggatgag ctctacaaag cagcgaacga cgaaaattac gcccttgcag cgtaaacgcg
     1741 tgctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1801 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagccg
     1861 tcttcacctc gagttaattt ttaaagtatg ggcaatcaat tgctcctgtt aaaattgctt
     1921 tagaaatact ttggcagcgg tttgttgtat tgagtttcat ttgcgcattg gttaaatgga
     1981 aagtgacagt acgctcactg cagcctaata tttttgaaat atcccaagag ctttttcctt
     2041 cgcatgccca cgctaaacat tctttttctc ttttggttaa atcgttgttt gatttattat
     2101 ttgctatatt tatttttcga taattatcaa ctagagaagg aacaattaat ggtatgttca
     2161 tacacgcatg taaaaataaa ctatctatat agttgtcttt ttctgaatgt gcaaaactaa
     2221 gcattccgaa gccattgtta gccgtatgaa tagggaaact aaacccagtg ataagacctg
     2281 atgttttcgc ttctttaatt acatttggag attttttatt tacagcattg ttttcaaata
     2341 tattccaatt aattggtgaa tgattggagt tagaataatc tactatagga tcatatttta
     2401 ttaaattagc gtcatcataa tattgcctcc attttttagg gtaattatct agaattgaaa
     2461 tatcagattt aaccatagaa tgaggataaa tgatcgcgag taaataatat tcacaatgta
     2521 ccattttagt catatcagat aagcattgat taatatcatt attgcttcta caagctttaa
     2581 ttttattaat tattctgtat gtgtcgtcgg catttatgtt tttcataccc atctctttat
     2641 ccttacctat tgtttgtcgc aagttttgcg tgttatatat cattaaaacg gtaatggatt
     2701 gacatttgat tctaataaat tggatttttg tcacactatt gtatcgctgg gaatacaatt
     2761 acttaacata agcacctgta ggatcgtaca ggtttacgca agaaaatggt ttgttatagt
     2821 cgaatgaatt cattaaagag gagaaaggta ccatgactat aatgataaaa aaatcggatt
     2881 ttttggcaat tccatcggag gagtataaag gtattctaag tcttcgttat caagtgttta
     2941 agcaaagact tgagtgggac ttagttgtag aaaataacct tgaatcagat gagtatgata
     3001 actcaaatgc agaatatatt tatgcttgtg atgatactga aaatgtaagt ggatgctggc
     3061 gtttattacc tacaacaggt gattatatgc tgaaaagtgt ttttcctgaa ttgcttggtc
     3121 aacagagtgc tcccaaagat cctaatatag tcgaattaag tcgttttgct gtaggtaaaa
     3181 atagctcaaa gataaataac tctgctagtg aaattacaat gaaactattt gaagctatat
     3241 ataaacacgc tgttagtcaa ggtattacag aatatgtaac agtaacatca acagcaatag
     3301 agcgattttt aaagcgtatt aaagttcctt gtcatcgtat tggagacaaa gaaattcatg
     3361 tattaggtga tactaaatcg gttgtattgt ctatgcctat taatgaacag tttaaaaaag
     3421 cagtcttaaa tgcagcgaac gacgaaaatt acgcccttgc agcgtaaacg cgtgctagag
     3481 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     3541 gtcggtgaac gctctcctga gtaggacaaa tccgccgccc tagacctagg gcgttcggct
     3601 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4141 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4381 ttaagggatt ttggtcatga ctagtgcttg gattctcacc aataaaaaac gcccggcggc
     4441 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     4501 ggagtccaag cgagctctcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     4561 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     4621 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     4681 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     4741 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     4801 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     4861 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     4921 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     4981 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     5041 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     5101 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     5161 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     5221 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     5281 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     5341 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     5401 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
     5461 caattccgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5521 cacgaggccc tttcgtcttc ac
//
