LOCUS       sequence-269559-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for quadruplet codon CGGC.
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: pED1a89
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..32
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(48..863)
                     /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(48..863)
                     /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     771..790
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     855..874
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(974..1924)
                     /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      1972..2194
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(2721..2738)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2891..2977
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3069..3096
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             3305..4385
                     /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
                     KLFQSDVMPFLKEKQRRPII"
     CDS             4449..5429
                     /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"
ORIGIN
        1 agtcaaaagc ctccgaccgg aggcttttga ctacatgccc atggcgttta gaaaaactca
       61 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
      121 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag aatggcaaga
      181 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
      241 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
      301 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
      361 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
      421 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
      481 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
      541 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
      601 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
      661 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
      721 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
      781 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgagcaagac
      841 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
      901 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgaagg
      961 ccaaataggc cgttcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt
     1021 gtttttgcgt gagccatgag aacgaaccat tgagatcatg cttactttgc atgtcactca
     1081 aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt
     1141 ttcttagtcc gttacgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat
     1201 tcatttttat ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa
     1261 cttggaaaat caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc
     1321 tgtaagtgtt taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact
     1381 catggtagtt attttcaagc attaacatga acttaaattc atcaaggcta atctctatat
     1441 ttgccttgtg agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag
     1501 agtatttgtt ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact
     1561 ggaaaagata aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact
     1621 tggcatagtt tgtccactgg aaaatccgaa agcctttaac caaaggattc ctgatttcca
     1681 cagttctcgt catcagctct ctggttgctt tagctaatac accataagca ttttccctac
     1741 tgatgttcat catctgagcg tattggttat aagtgaacga taccgtccgt tctttccttg
     1801 tagggttttc aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat
     1861 gctccgttaa gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag
     1921 acatacatct caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc
     1981 aatgataatt actagtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct
     2041 ttgctggaaa acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg
     2101 ttttttttgt ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga
     2161 taaaaagaat agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt
     2221 acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag
     2281 gcttaagtag caccctcgca agctcgggca aatcgctgaa tattcctttt gtctccgacc
     2341 atcaggcacc tgagtcgctg tctttttcgt gacattcagt tcgctgcgct cacggctctg
     2401 gcagtgaatg ggggtaaatg gcactacagg cgccttttat ggattcatgc aaggaaacta
     2461 cccataatac aagaaaagcc cgtcacgggc ttctcagggc gttttatggc gggtctgcta
     2521 tgtggtgcta tctgactttt tgctgttcag cagttcctgc cctctgattt tccagtctga
     2581 ccacttcgga ttatcccgtg acaggtcatt cagactggct aatgcaccca gtaaggcagc
     2641 ggtatcatca acaggcttac ccgtcttact gtcaagagga catccggtgg ctgttttggc
     2701 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     2761 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     2821 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     2881 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     2941 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     3001 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     3061 aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcgctgagg
     3121 attctagagc acagctaaca ccacgtcgtc cctatctgct gccctaggtc tatgagtggt
     3181 tgctggataa ctttacgggc atgcataagg ctcgtataat atattcaggg agaccacaac
     3241 ggtttccctc tacaaataat tttgtttaac tttatatctg ttatttttta aggaggaaaa
     3301 aaaaatgaaa tttggaaact ttttgcttac ataccaacct ccccaatttt cccaaacaga
     3361 ggtaatgaaa cgtttggtta aattaggtcg catctctgag gagtgtggtt ttgataccgt
     3421 atggttactg gagcatcatt tcacggagtt tggtttgctt ggtaaccctt atgtcgctgc
     3481 tgcatattta cttggcgcga ctaaaaaatt gaatgtagga actgccgcta ttgttcttcc
     3541 cacagcccat ccagtacgcc aacttgaaga tgtgaattta ttggatcaaa tgtcaaaagg
     3601 acgatttcgg tttggtattt gccgagggct ttacaacaag gactttcgcg tattcggaac
     3661 agatatgaat aacagtcgcg ccttagcgga atgctggtac gggctgataa agaatggcat
     3721 gacagaggga tatatggaag ctgataatga acatatcaag ttccataagg taaaagtaaa
     3781 ccccgcggcg tatagcagag gtggcgcacc ggtttatgtg gtggctgaat cagcttcgac
     3841 gactgagtgg gctgctcaat ttggcctacc gatgatatta agttggatta taaatactaa
     3901 cgaaaagaaa gcacaacttg agctttataa tgaagtggct caagaatatg ggcacgatat
     3961 tcataatatc gaccattgct tatcatatat aacatctgta gatcatgact caattaaagc
     4021 gaaagagatt tgccggaaat ttctggggca ttggtatgat tcttatgtga atgctacgac
     4081 tatttttgat gattcagacc aaacaagagg ttatgatttc aataaagggc agtggcgtga
     4141 ctttgtatta aaaggacata aagatactaa tcgccgtatt gattacagtt acgaaatcaa
     4201 tcccgtggga acgccgcagg aatgtattga cataattcaa aaagacattg atgctacagg
     4261 aatatcaaat atttgttgtg gatttgaagc taatggaaca gtagacgaaa ttattgcttc
     4321 catgaagctc ttccagtctg atgtcatgcc atttcttaaa gaaaaacaac gtcggcctat
     4381 tatattatgg cggtggcggt agcggcggtg gcggtagcgg cggtggcggt agcggcggtg
     4441 gcggtagcaa atttggattg ttcttcctta acttcatcaa ttcaacaact gttcaagaac
     4501 agagtatagt tcgcatgcag gaaataacgg agtatgttga taagttgaat tttgaacaga
     4561 ttttagtgta tgaaaatcat ttttcagata atggtgttgt cggcgctcct ctgactgttt
     4621 ctggttttct gctcggttta acagagaaaa ttaaaattgg ttcattaaat cacatcatta
     4681 caactcatca tcctgtccgc atagcggagg aagcttgctt attggatcag ttaagtgaag
     4741 ggagatttat tttagggttt agtgattgcg aaaaaaaaga tgaaatgcat ttttttaatc
     4801 gcccggttga atatcaacag caactatttg aagagtgtta tgaaatcatt aacgatgctt
     4861 taacaacagg ctattgtaat ccagataacg atttttatag cttccctaaa atatctgtaa
     4921 atccccatgc ttatacgcca ggcggacctc ggaaatatgt aacagcaacc agtcatcata
     4981 ttgttgagtg ggcggccaaa aaaggtattc ctctcatctt taagtgggat gattctaatg
     5041 atgttagata tgaatatgct gaaagatata aagccgttgc ggataaatat gacgttgacc
     5101 tatcagagat agaccatcag ttaatgatat tagttaacta taacgaagat agtaataaag
     5161 ctaaacaaga gacgcgtgca tttattagtg attatgttct tgaaatgcac cctaatgaaa
     5221 atttcgaaaa taaacttgaa gaaataattg cagaaaacgc tgtcggaaat tatacggagt
     5281 gtataactgc ggctaagttg gcaattgaaa agtgtggtgc gaaaagtgta ttgctgtcct
     5341 ttgaaccaat gaatgatttg atgagccaaa aaaatgtaat caatattgtt gatgataata
     5401 ttaagaagta ccacacggaa tatacctaaa cttaattaac ggcactcctc agcca
//
