LOCUS       sequence-307530-        6064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses orthogonal firefly luciferase gene in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  AUTHORS   Aleksashin NA, Szal T, d'Aquino AE, Jewett MC, Vazquez-Laslop N, 
            Mankin AS
  TITLE     A fully orthogonal system for protein synthesis in bacterial cells.
  JOURNAL   Nat Commun. 2020 Apr 20;11(1):1858. doi: 10.1038/s41467-020-15756-1.
  PUBMED    32313034
REFERENCE   2  (bases 1 to 6064)
  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"; journalName: "Nat 
            Commun."; date: "2020-04-20"; volume: "11(1):1858. doi"; pages: " 
            10.1038/s41467-020-15756-1"
COMMENT     Sequence Label: poLuc1
FEATURES             Location/Qualifiers
     source          1..6064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             63..74
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      878..949
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      965..992
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1094..2746
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1160..1178)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(join(3163..3168,3169..3185,3186..3191))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3303..3848
                     /direction=RIGHT
                     /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     3749..3768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3965..3982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             5035..5826
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLAPRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcactactag
       61 agaaagagga gaaatactag atgaaaaaca taaatgccga cgacacatac agaataatta
      121 ataaaattaa agcttgtaga agcaataatg atattaatca atgcttatct gatatgacta
      181 aaatggtaca ttgtgaatat tatttactcg cgatcattta tcctcattct atggttaaat
      241 ctgatatttc aatcctagat aattacccta aaaaatggag gcaatattat gatgacgcta
      301 atttaataaa atatgatcct atagtagatt attctaactc caatcattca ccaattaatt
      361 ggaatatatt tgaaaacaat gctgtaaata aaaaatctcc aaatgtaatt aaagaagcga
      421 aaacatcagg tcttatcact gggtttagtt tccctattca tacggctaac aatggcttcg
      481 gaatgcttag ttttgcacat tcagaaaaag acaactatat agatagttta tttttacatg
      541 cgtgtatgaa cataccatta attgttcctt ctctagttga taattatcga aaaataaata
      601 tagcaaataa taaatcaaac aacgatttaa ccaaaagaga aaaagaatgt ttagcgtggg
      661 catgcgaagg aaaaagctct tgggatattt caaaaatatt aggttgcagt gagcgtactg
      721 tcactttcca tttaaccaat gcgcaaatga aactcaatac aacaaaccgc tgccaaagta
      781 tttctaaagc aattttaaca ggagcaattg attgcccata ctttaaaaat taataacact
      841 gatagtgcta gtgtagatca ctactagagc caggcatcaa ataaaacgaa aggctcagtc
      901 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
      961 cactggctca ccttcgggtg ggcctttctg cgtttatata ctagagacct gtaggatcgt
     1021 acaggtttac gcaagaaaat ggtttgttat agtcgaataa actatatctg ttattttttc
     1081 caaccacaga tctatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc
     1141 tctagaggat ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt
     1201 tcctggaaca attgctttta cagatgcaca tatcgaggtg aacatcacgt acgcggaata
     1261 cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca
     1321 cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt
     1381 atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa
     1441 cagtatgaac atttcgcagc ctaccgtagt gtttgtttcc aaaaaggggt tgcaaaaaat
     1501 tttgaacgtg caaaaaaaat taccaataat ccagaaaatt attatcatgg attctaaaac
     1561 ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt
     1621 taatgaatac gattttgtac cagagtcctt tgatcgtgac aaaacaattg cactgataat
     1681 gaattcctct ggatctactg ggttacctaa gggtgtggcc cttccgcata gaactgcctg
     1741 cgtcagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc
     1801 gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt
     1861 gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttttacgatc
     1921 ccttcaggat tacaaaattc aaagtgcgtt gctagtacca accctatttt cattcttcgc
     1981 caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg cttctggggg
     2041 cgcacctctt tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc ttccagggat
     2101 acgacaagga tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga
     2161 tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct
     2221 ggataccggg aaaacgctgg gcgttaatca gagaggcgaa ttatgtgtca gaggacctat
     2281 gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg
     2341 atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct tcatagttga
     2401 ccgcttgaag tctttaatta aatacaaagg atatcaggtg gcccccgctg aattggaatc
     2461 gatattgtta caacacccca acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga
     2521 cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa
     2581 agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt
     2641 tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag
     2701 agagatcctc ataaaggcca agaagggcgg aaagtccaaa ttgtaagtcg acgggcataa
     2761 ataggtttaa ttttgctacg ggggcgttat ttaggttttt tcttctttcg aaaaaatctt
     2821 tctttatgaa gttaaaagct atgtattcaa tagcatattt tgaatatgga catagaatag
     2881 tgcttatcac tattgcatat agcatcttat ctgacacaag gaaataatac ccttcgctgt
     2941 tttttgttat aaggtatata tatataagtg tgcagtacag gccaaataaa atatttttta
     3001 tgtagtatct taaatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     3061 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     3121 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     3181 taagctgtca aacatgagaa ttacgactta tatcgtatgg ggctgacttc aggtgctaca
     3241 tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct
     3301 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     3361 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     3421 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     3481 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     3541 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     3601 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     3661 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     3721 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     3781 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     3841 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     3901 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     3961 gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     4021 caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     4081 ccaacatagt aagccagtat acactccgct agcgctgatg tccggcggtg cttttgccgt
     4141 tacgcaccac cccgtcagta gctgaacagg agggacagag ctttatgctt gtaaaccgtt
     4201 ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg gtccccaatt aattagtaat
     4261 ataatctatt aaaggtcatt caaaaggtca tccaccggat caattcccct gctcgcgcag
     4321 gctgggtgcc aagctctcgg gtaacatcaa ggcccgatcc ttggagccct tgccctcccg
     4381 cacgatgatc gtgccgtgat cgaaatccag atccttgacc cgcagttgca aaccctcact
     4441 gatccgcatg gggcccccgt tccatacaga agctgggcga acaaacgatg ctcgccttcc
     4501 agaaaaccga ggatgcgaac cacttcatcc ggggtcagca ccaccggcaa gcgccgcgac
     4561 ggccgaggtc ttccgatctc ctgaagccag ggcagatccg tgcacagcac cttgccgtag
     4621 aagaacagca aggccgccaa tgcctgacga tgcgtggaga ccgaaacctt gcgctcgttc
     4681 gccagccagg acagaaatgc ctcgacttcg ctgctgccca aggttgccgg gtgacgcaca
     4741 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
     4801 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     4861 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
     4921 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     4981 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgagg
     5041 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
     5101 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     5161 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     5221 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     5281 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     5341 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     5401 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     5461 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     5521 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     5581 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     5641 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     5701 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggccccgcgc
     5761 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     5821 aaataatgtc taacaattcg ttcaagccga cgccgcttcg cggcgcggct taactcaagc
     5881 gttagatgca ctaagcacat aattgctcac agccaaacta tcaggtcaag tctgctttta
     5941 ttatttttaa gcgtgcataa taagccctac acaaattggg agatatatca tgaaaggctg
     6001 gctttttctt gttatcgcaa tagttggcga agtaatcgca acatattcgc ggccgcttct
     6061 agag
//
