LOCUS       sequence-269560-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for quadruplet codon AGGA.
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: pED1a71
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3..983
                     /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      1010..1041
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(1057..1872)
                     /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(1057..1872)
                     /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     1780..1799
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1864..1883
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1983..2933)
                     /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      2981..3203
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(3730..3747)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3900..3986
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4078..4105
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             4314..5394
                     /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
                     KLFQSDVMPFLKEKQRRTII"
ORIGIN
        1 gcaaatttgg attgttcttc cttaacttca tcaattcaac aactgttcaa gaacagagta
       61 tagttcgcat gcaggaaata acggagtatg ttgataagtt gaattttgaa cagattttag
      121 tgtatgaaaa tcatttttca gataatggtg ttgtcggcgc tcctctgact gtttctggtt
      181 ttctgctcgg tttaacagag aaaattaaaa ttggttcatt aaatcacatc attacaactc
      241 atcatcctgt ccgcatagcg gaggaagctt gcttattgga tcagttaagt gaagggagat
      301 ttattttagg gtttagtgat tgcgaaaaaa aagatgaaat gcattttttt aatcgcccgg
      361 ttgaatatca acagcaacta tttgaagagt gttatgaaat cattaacgat gctttaacaa
      421 caggctattg taatccagat aacgattttt atagcttccc taaaatatct gtaaatcccc
      481 atgcttatac gccaggcgga cctcggaaat atgtaacagc aaccagtcat catattgttg
      541 agtgggcggc caaaaaaggt attcctctca tctttaagtg ggatgattct aatgatgtta
      601 gatatgaata tgctgaaaga tataaagccg ttgcggataa atatgacgtt gacctatcag
      661 agatagacca tcagttaatg atattagtta actataacga agatagtaat aaagctaaac
      721 aagagacgcg tgcatttatt agtgattatg ttcttgaaat gcaccctaat gaaaatttcg
      781 aaaataaact tgaagaaata attgcagaaa acgctgtcgg aaattatacg gagtgtataa
      841 ctgcggctaa gttggcaatt gaaaagtgtg gtgcgaaaag tgtattgctg tcctttgaac
      901 caatgaatga tttgatgagc caaaaaaatg taatcaatat tgttgatgat aatattaaga
      961 agtaccacac ggaatatacc taaacttaat taacggcact cctcagccaa gtcaaaagcc
     1021 tccgaccgga ggcttttgac tacatgccca tggcgtttag aaaaactcat cgagcatcaa
     1081 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1141 ctgtaatgaa ggagaaaact caccgaggca gttccataga atggcaagat cctggtatcg
     1201 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     1261 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     1321 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     1381 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     1441 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     1501 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     1561 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     1621 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     1681 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     1741 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     1801 atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg
     1861 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     1921 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgaaggc caaataggcc
     1981 gttcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg tttttgcgtg
     2041 agccatgaga acgaaccatt gagatcatgc ttactttgca tgtcactcaa aaattttgcc
     2101 tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg
     2161 ttacgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt catttttatc
     2221 tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac ttggaaaatc
     2281 aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct gtaagtgttt
     2341 aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc atggtagtta
     2401 ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt tgccttgtga
     2461 gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga gtatttgttt
     2521 tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg gaaaagataa
     2581 ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt ggcatagttt
     2641 gtccactgga aaatccgaaa gcctttaacc aaaggattcc tgatttccac agttctcgtc
     2701 atcagctctc tggttgcttt agctaataca ccataagcat tttccctact gatgttcatc
     2761 atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt agggttttca
     2821 atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg ctccgttaag
     2881 tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga catacatctc
     2941 aattggtcta ggtgatttta atcactatac caattgagat gggctagtca atgataatta
     3001 ctagtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt tgctggaaaa
     3061 cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt
     3121 tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat aaaaagaata
     3181 gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta caaaaggatg
     3241 tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg cttaagtagc
     3301 accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca tcaggcacct
     3361 gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg
     3421 gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac ccataataca
     3481 agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat
     3541 ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat
     3601 tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa
     3661 caggcttacc cgtcttactg tcaagaggac atccggtggc tgttttggcg gatgagagaa
     3721 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     3781 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     3841 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     3901 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     3961 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     4021 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     4081 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcgctgagga ttctagagca
     4141 cagctaacac cacgtcgtcc ctatctgctg ccctaggtct atgagtggtt gctggataac
     4201 tttacgggca tgcataaggc tcgtataata tattcaggga gaccacaacg gtttccctct
     4261 acaaataatt ttgtttaact ttatatctgt tattttttaa ggaggaaaaa aaaatgaaat
     4321 ttggaaactt tttgcttaca taccaacctc cccaattttc ccaaacagag gtaatgaaac
     4381 gtttggttaa attaggtcgc atctctgagg agtgtggttt tgataccgta tggttactgg
     4441 agcatcattt cacggagttt ggtttgcttg gtaaccctta tgtcgctgct gcatatttac
     4501 ttggcgcgac taaaaaattg aatgtaggaa ctgccgctat tgttcttccc acagcccatc
     4561 cagtacgcca acttgaagat gtgaatttat tggatcaaat gtcaaaagga cgatttcggt
     4621 ttggtatttg ccgagggctt tacaacaagg actttcgcgt attcggaaca gatatgaata
     4681 acagtcgcgc cttagcggaa tgctggtacg ggctgataaa gaatggcatg acagagggat
     4741 atatggaagc tgataatgaa catatcaagt tccataaggt aaaagtaaac cccgcggcgt
     4801 atagcagagg tggcgcaccg gtttatgtgg tggctgaatc agcttcgacg actgagtggg
     4861 ctgctcaatt tggcctaccg atgatattaa gttggattat aaatactaac gaaaagaaag
     4921 cacaacttga gctttataat gaagtggctc aagaatatgg gcacgatatt cataatatcg
     4981 accattgctt atcatatata acatctgtag atcatgactc aattaaagcg aaagagattt
     5041 gccggaaatt tctggggcat tggtatgatt cttatgtgaa tgctacgact atttttgatg
     5101 attcagacca aacaagaggt tatgatttca ataaagggca gtggcgtgac tttgtattaa
     5161 aaggacataa agatactaat cgccgtattg attacagtta cgaaatcaat cccgtgggaa
     5221 cgccgcagga atgtattgac ataattcaaa aagacattga tgctacagga atatcaaata
     5281 tttgttgtgg atttgaagct aatggaacag tagacgaaat tattgcttcc atgaagctct
     5341 tccagtctga tgtcatgcca tttcttaaag aaaaacaacg taggactatt atattatggc
     5401 ggtggcggta gcggcggtgg cggtagcggc ggtggcggta gcggcggtgg cggta
//
