LOCUS       sequence-259777-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for quadruplet codon CGGT.
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: pED1a36
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     291..310
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     375..394
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(494..1444)
                     /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      1492..1714
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(2241..2258)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2411..2497
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2589..2616
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2825..3905
                     /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
                     KLFQSDVMPFLKEKQRRSII"
     CDS             3969..4949
                     /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      4976..5007
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(join(5023..5455,1..383))
                     /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(join(5023..5455,1..383))
                     /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"
ORIGIN
        1 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
       61 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
      121 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
      181 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
      241 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
      301 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgagcaagac
      361 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
      421 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgaagg
      481 ccaaataggc cgttcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt
      541 gtttttgcgt gagccatgag aacgaaccat tgagatcatg cttactttgc atgtcactca
      601 aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt
      661 ttcttagtcc gttacgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat
      721 tcatttttat ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa
      781 cttggaaaat caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc
      841 tgtaagtgtt taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact
      901 catggtagtt attttcaagc attaacatga acttaaattc atcaaggcta atctctatat
      961 ttgccttgtg agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag
     1021 agtatttgtt ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact
     1081 ggaaaagata aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact
     1141 tggcatagtt tgtccactgg aaaatccgaa agcctttaac caaaggattc ctgatttcca
     1201 cagttctcgt catcagctct ctggttgctt tagctaatac accataagca ttttccctac
     1261 tgatgttcat catctgagcg tattggttat aagtgaacga taccgtccgt tctttccttg
     1321 tagggttttc aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat
     1381 gctccgttaa gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag
     1441 acatacatct caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc
     1501 aatgataatt actagtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct
     1561 ttgctggaaa acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg
     1621 ttttttttgt ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga
     1681 taaaaagaat agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt
     1741 acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag
     1801 gcttaagtag caccctcgca agctcgggca aatcgctgaa tattcctttt gtctccgacc
     1861 atcaggcacc tgagtcgctg tctttttcgt gacattcagt tcgctgcgct cacggctctg
     1921 gcagtgaatg ggggtaaatg gcactacagg cgccttttat ggattcatgc aaggaaacta
     1981 cccataatac aagaaaagcc cgtcacgggc ttctcagggc gttttatggc gggtctgcta
     2041 tgtggtgcta tctgactttt tgctgttcag cagttcctgc cctctgattt tccagtctga
     2101 ccacttcgga ttatcccgtg acaggtcatt cagactggct aatgcaccca gtaaggcagc
     2161 ggtatcatca acaggcttac ccgtcttact gtcaagagga catccggtgg ctgttttggc
     2221 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     2281 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     2341 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     2401 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     2461 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     2521 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     2581 aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcgctgagg
     2641 attctagagc acagctaaca ccacgtcgtc cctatctgct gccctaggtc tatgagtggt
     2701 tgctggataa ctttacgggc atgcataagg ctcgtataat atattcaggg agaccacaac
     2761 ggtttccctc tacaaataat tttgtttaac tttatatctg ttatttttta aggaggaaaa
     2821 aaaaatgaaa tttggaaact ttttgcttac ataccaacct ccccaatttt cccaaacaga
     2881 ggtaatgaaa cgtttggtta aattaggtcg catctctgag gagtgtggtt ttgataccgt
     2941 atggttactg gagcatcatt tcacggagtt tggtttgctt ggtaaccctt atgtcgctgc
     3001 tgcatattta cttggcgcga ctaaaaaatt gaatgtagga actgccgcta ttgttcttcc
     3061 cacagcccat ccagtacgcc aacttgaaga tgtgaattta ttggatcaaa tgtcaaaagg
     3121 acgatttcgg tttggtattt gccgagggct ttacaacaag gactttcgcg tattcggaac
     3181 agatatgaat aacagtcgcg ccttagcgga atgctggtac gggctgataa agaatggcat
     3241 gacagaggga tatatggaag ctgataatga acatatcaag ttccataagg taaaagtaaa
     3301 ccccgcggcg tatagcagag gtggcgcacc ggtttatgtg gtggctgaat cagcttcgac
     3361 gactgagtgg gctgctcaat ttggcctacc gatgatatta agttggatta taaatactaa
     3421 cgaaaagaaa gcacaacttg agctttataa tgaagtggct caagaatatg ggcacgatat
     3481 tcataatatc gaccattgct tatcatatat aacatctgta gatcatgact caattaaagc
     3541 gaaagagatt tgccggaaat ttctggggca ttggtatgat tcttatgtga atgctacgac
     3601 tatttttgat gattcagacc aaacaagagg ttatgatttc aataaagggc agtggcgtga
     3661 ctttgtatta aaaggacata aagatactaa tcgccgtatt gattacagtt acgaaatcaa
     3721 tcccgtggga acgccgcagg aatgtattga cataattcaa aaagacattg atgctacagg
     3781 aatatcaaat atttgttgtg gatttgaagc taatggaaca gtagacgaaa ttattgcttc
     3841 catgaagctc ttccagtctg atgtcatgcc atttcttaaa gaaaaacaac gtcggtctat
     3901 tatattatgg cggtggcggt agcggcggtg gcggtagcgg cggtggcggt agcggcggtg
     3961 gcggtagcaa atttggattg ttcttcctta acttcatcaa ttcaacaact gttcaagaac
     4021 agagtatagt tcgcatgcag gaaataacgg agtatgttga taagttgaat tttgaacaga
     4081 ttttagtgta tgaaaatcat ttttcagata atggtgttgt cggcgctcct ctgactgttt
     4141 ctggttttct gctcggttta acagagaaaa ttaaaattgg ttcattaaat cacatcatta
     4201 caactcatca tcctgtccgc atagcggagg aagcttgctt attggatcag ttaagtgaag
     4261 ggagatttat tttagggttt agtgattgcg aaaaaaaaga tgaaatgcat ttttttaatc
     4321 gcccggttga atatcaacag caactatttg aagagtgtta tgaaatcatt aacgatgctt
     4381 taacaacagg ctattgtaat ccagataacg atttttatag cttccctaaa atatctgtaa
     4441 atccccatgc ttatacgcca ggcggacctc ggaaatatgt aacagcaacc agtcatcata
     4501 ttgttgagtg ggcggccaaa aaaggtattc ctctcatctt taagtgggat gattctaatg
     4561 atgttagata tgaatatgct gaaagatata aagccgttgc ggataaatat gacgttgacc
     4621 tatcagagat agaccatcag ttaatgatat tagttaacta taacgaagat agtaataaag
     4681 ctaaacaaga gacgcgtgca tttattagtg attatgttct tgaaatgcac cctaatgaaa
     4741 atttcgaaaa taaacttgaa gaaataattg cagaaaacgc tgtcggaaat tatacggagt
     4801 gtataactgc ggctaagttg gcaattgaaa agtgtggtgc gaaaagtgta ttgctgtcct
     4861 ttgaaccaat gaatgatttg atgagccaaa aaaatgtaat caatattgtt gatgataata
     4921 ttaagaagta ccacacggaa tatacctaaa cttaattaac ggcactcctc agccaagtca
     4981 aaagcctccg accggaggct tttgactaca tgcccatggc gtttagaaaa actcatcgag
     5041 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     5101 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     5161 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     5221 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     5281 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     5341 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     5401 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgc
//
