LOCUS       sequence-263777-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for quadruplet codon TAGA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5455)
  AUTHORS   DeBenedictis EA, Carver GD, Chung CZ, Soll D, Badran AH
  TITLE     Multiplex suppression of four quadruplet codons via tRNA directed 
            evolution.
  JOURNAL   Nat Commun. 2021 Sep 29;12(1):5706. doi: 10.1038/s41467-021-25948-y.
  PUBMED    34588441
REFERENCE   2  (bases 1 to 5455)
  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: "2021-09-29"; volume: "12(1):5706. doi"; pages: " 
            10.1038/s41467-021-25948-y"
COMMENT     Sequence Label: pED1a6
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      109..195
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      287..314
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             523..1603
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA luciferase subunit"
                     /label=LuxA
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQR*TII"
     CDS             1667..2647
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB luciferase subunit"
                     /label=LuxB
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="KFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENHF
                     SDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILGF
                     SDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAYT
                     PGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEID
                     HQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECITA
                     AKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHTEYT"
     terminator      2674..2705
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2721..3536)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2721..3536)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3444..3463
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3528..3547
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3647..4597)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFRIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4645..4867
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(5394..5411)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
       61 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
      121 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
      181 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
      241 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
      301 acggatggcc tttttgcgtt tctacaaact cgctgaggat tctagagcac agctaacacc
      361 acgtcgtccc tatctgctgc cctaggtcta tgagtggttg ctggataact ttacgggcat
      421 gcataaggct cgtataatat attcagggag accacaacgg tttccctcta caaataattt
      481 tgtttaactt tatatctgtt attttttaag gaggaaaaaa aaatgaaatt tggaaacttt
      541 ttgcttacat accaacctcc ccaattttcc caaacagagg taatgaaacg tttggttaaa
      601 ttaggtcgca tctctgagga gtgtggtttt gataccgtat ggttactgga gcatcatttc
      661 acggagtttg gtttgcttgg taacccttat gtcgctgctg catatttact tggcgcgact
      721 aaaaaattga atgtaggaac tgccgctatt gttcttccca cagcccatcc agtacgccaa
      781 cttgaagatg tgaatttatt ggatcaaatg tcaaaaggac gatttcggtt tggtatttgc
      841 cgagggcttt acaacaagga ctttcgcgta ttcggaacag atatgaataa cagtcgcgcc
      901 ttagcggaat gctggtacgg gctgataaag aatggcatga cagagggata tatggaagct
      961 gataatgaac atatcaagtt ccataaggta aaagtaaacc ccgcggcgta tagcagaggt
     1021 ggcgcaccgg tttatgtggt ggctgaatca gcttcgacga ctgagtgggc tgctcaattt
     1081 ggcctaccga tgatattaag ttggattata aatactaacg aaaagaaagc acaacttgag
     1141 ctttataatg aagtggctca agaatatggg cacgatattc ataatatcga ccattgctta
     1201 tcatatataa catctgtaga tcatgactca attaaagcga aagagatttg ccggaaattt
     1261 ctggggcatt ggtatgattc ttatgtgaat gctacgacta tttttgatga ttcagaccaa
     1321 acaagaggtt atgatttcaa taaagggcag tggcgtgact ttgtattaaa aggacataaa
     1381 gatactaatc gccgtattga ttacagttac gaaatcaatc ccgtgggaac gccgcaggaa
     1441 tgtattgaca taattcaaaa agacattgat gctacaggaa tatcaaatat ttgttgtgga
     1501 tttgaagcta atggaacagt agacgaaatt attgcttcca tgaagctctt ccagtctgat
     1561 gtcatgccat ttcttaaaga aaaacaacgt tagactatta tattatggcg gtggcggtag
     1621 cggcggtggc ggtagcggcg gtggcggtag cggcggtggc ggtagcaaat ttggattgtt
     1681 cttccttaac ttcatcaatt caacaactgt tcaagaacag agtatagttc gcatgcagga
     1741 aataacggag tatgttgata agttgaattt tgaacagatt ttagtgtatg aaaatcattt
     1801 ttcagataat ggtgttgtcg gcgctcctct gactgtttct ggttttctgc tcggtttaac
     1861 agagaaaatt aaaattggtt cattaaatca catcattaca actcatcatc ctgtccgcat
     1921 agcggaggaa gcttgcttat tggatcagtt aagtgaaggg agatttattt tagggtttag
     1981 tgattgcgaa aaaaaagatg aaatgcattt ttttaatcgc ccggttgaat atcaacagca
     2041 actatttgaa gagtgttatg aaatcattaa cgatgcttta acaacaggct attgtaatcc
     2101 agataacgat ttttatagct tccctaaaat atctgtaaat ccccatgctt atacgccagg
     2161 cggacctcgg aaatatgtaa cagcaaccag tcatcatatt gttgagtggg cggccaaaaa
     2221 aggtattcct ctcatcttta agtgggatga ttctaatgat gttagatatg aatatgctga
     2281 aagatataaa gccgttgcgg ataaatatga cgttgaccta tcagagatag accatcagtt
     2341 aatgatatta gttaactata acgaagatag taataaagct aaacaagaga cgcgtgcatt
     2401 tattagtgat tatgttcttg aaatgcaccc taatgaaaat ttcgaaaata aacttgaaga
     2461 aataattgca gaaaacgctg tcggaaatta tacggagtgt ataactgcgg ctaagttggc
     2521 aattgaaaag tgtggtgcga aaagtgtatt gctgtccttt gaaccaatga atgatttgat
     2581 gagccaaaaa aatgtaatca atattgttga tgataatatt aagaagtacc acacggaata
     2641 tacctaaact taattaacgg cactcctcag ccaagtcaaa agcctccgac cggaggcttt
     2701 tgactacatg cccatggcgt ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     2761 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     2821 aactcaccga ggcagttcca tagaatggca agatcctggt atcggtctgc gattccgact
     2881 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     2941 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     3001 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     3061 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     3121 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     3181 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     3241 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     3301 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     3361 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     3421 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     3481 atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcataaca
     3541 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     3601 tcttgtgcaa tgtaacatca gagattttga aggccaaata ggccgttcag atccttccgt
     3661 atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac
     3721 cattgagatc atgcttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct
     3781 gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttacgt aggtaggaat
     3841 ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt
     3901 cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc
     3961 ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg
     4021 gtttcaaaac ccattggtta agccttttaa actcatggta gttattttca agcattaaca
     4081 tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta
     4141 gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat
     4201 gttccagatt atattttatg aattttttta actggaaaag ataaggcaat atctcttcac
     4261 taaaaactaa ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatcc
     4321 gaaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg
     4381 ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt
     4441 tataagtgaa cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta
     4501 gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg
     4561 ctagttcatt tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat
     4621 tttaatcact ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt
     4681 gagttgtggg tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta
     4741 gaccctctgt aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta
     4801 taatttatag aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt
     4861 ataactcact actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg
     4921 ctcctctaca aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg
     4981 gcaaatcgct gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt
     5041 cgtgacattc agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac
     5101 aggcgccttt tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg
     5161 ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt
     5221 cagcagttcc tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc
     5281 attcagactg gctaatgcac ccagtaaggc agcggtatca tcaacaggct tacccgtctt
     5341 actgtcaaga ggacatccgg tggctgtttt ggcggatgag agaagatttt cagcctgata
     5401 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggca
//
