LOCUS       sequence-263779-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for quadruplet codon TACA.
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: pED1a39
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..87
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      179..206
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             415..1495
                     /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
                     KLFQSDVMPFLKEKQRYTII"
     CDS             1559..2539
                     /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      2566..2597
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2613..3428)
                     /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(2613..3428)
                     /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     3336..3355
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3420..3439
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3539..4489)
                     /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      4537..4759
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(5286..5303)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
       61 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
      121 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
      181 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcgctgagg attctagagc
      241 acagctaaca ccacgtcgtc cctatctgct gccctaggtc tatgagtggt tgctggataa
      301 ctttacgggc atgcataagg ctcgtataat atattcaggg agaccacaac ggtttccctc
      361 tacaaataat tttgtttaac tttatatctg ttatttttta aggaggaaaa aaaaatgaaa
      421 tttggaaact ttttgcttac ataccaacct ccccaatttt cccaaacaga ggtaatgaaa
      481 cgtttggtta aattaggtcg catctctgag gagtgtggtt ttgataccgt atggttactg
      541 gagcatcatt tcacggagtt tggtttgctt ggtaaccctt atgtcgctgc tgcatattta
      601 cttggcgcga ctaaaaaatt gaatgtagga actgccgcta ttgttcttcc cacagcccat
      661 ccagtacgcc aacttgaaga tgtgaattta ttggatcaaa tgtcaaaagg acgatttcgg
      721 tttggtattt gccgagggct ttacaacaag gactttcgcg tattcggaac agatatgaat
      781 aacagtcgcg ccttagcgga atgctggtac gggctgataa agaatggcat gacagaggga
      841 tatatggaag ctgataatga acatatcaag ttccataagg taaaagtaaa ccccgcggcg
      901 tatagcagag gtggcgcacc ggtttatgtg gtggctgaat cagcttcgac gactgagtgg
      961 gctgctcaat ttggcctacc gatgatatta agttggatta taaatactaa cgaaaagaaa
     1021 gcacaacttg agctttataa tgaagtggct caagaatatg ggcacgatat tcataatatc
     1081 gaccattgct tatcatatat aacatctgta gatcatgact caattaaagc gaaagagatt
     1141 tgccggaaat ttctggggca ttggtatgat tcttatgtga atgctacgac tatttttgat
     1201 gattcagacc aaacaagagg ttatgatttc aataaagggc agtggcgtga ctttgtatta
     1261 aaaggacata aagatactaa tcgccgtatt gattacagtt acgaaatcaa tcccgtggga
     1321 acgccgcagg aatgtattga cataattcaa aaagacattg atgctacagg aatatcaaat
     1381 atttgttgtg gatttgaagc taatggaaca gtagacgaaa ttattgcttc catgaagctc
     1441 ttccagtctg atgtcatgcc atttcttaaa gaaaaacaac gttacactat tatattatgg
     1501 cggtggcggt agcggcggtg gcggtagcgg cggtggcggt agcggcggtg gcggtagcaa
     1561 atttggattg ttcttcctta acttcatcaa ttcaacaact gttcaagaac agagtatagt
     1621 tcgcatgcag gaaataacgg agtatgttga taagttgaat tttgaacaga ttttagtgta
     1681 tgaaaatcat ttttcagata atggtgttgt cggcgctcct ctgactgttt ctggttttct
     1741 gctcggttta acagagaaaa ttaaaattgg ttcattaaat cacatcatta caactcatca
     1801 tcctgtccgc atagcggagg aagcttgctt attggatcag ttaagtgaag ggagatttat
     1861 tttagggttt agtgattgcg aaaaaaaaga tgaaatgcat ttttttaatc gcccggttga
     1921 atatcaacag caactatttg aagagtgtta tgaaatcatt aacgatgctt taacaacagg
     1981 ctattgtaat ccagataacg atttttatag cttccctaaa atatctgtaa atccccatgc
     2041 ttatacgcca ggcggacctc ggaaatatgt aacagcaacc agtcatcata ttgttgagtg
     2101 ggcggccaaa aaaggtattc ctctcatctt taagtgggat gattctaatg atgttagata
     2161 tgaatatgct gaaagatata aagccgttgc ggataaatat gacgttgacc tatcagagat
     2221 agaccatcag ttaatgatat tagttaacta taacgaagat agtaataaag ctaaacaaga
     2281 gacgcgtgca tttattagtg attatgttct tgaaatgcac cctaatgaaa atttcgaaaa
     2341 taaacttgaa gaaataattg cagaaaacgc tgtcggaaat tatacggagt gtataactgc
     2401 ggctaagttg gcaattgaaa agtgtggtgc gaaaagtgta ttgctgtcct ttgaaccaat
     2461 gaatgatttg atgagccaaa aaaatgtaat caatattgtt gatgataata ttaagaagta
     2521 ccacacggaa tatacctaaa cttaattaac ggcactcctc agccaagtca aaagcctccg
     2581 accggaggct tttgactaca tgcccatggc gtttagaaaa actcatcgag catcaaatga
     2641 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     2701 aatgaaggag aaaactcacc gaggcagttc catagaatgg caagatcctg gtatcggtct
     2761 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     2821 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta
     2881 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     2941 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     3001 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     3061 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     3121 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     3181 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     3241 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     3301 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     3361 aaatcagcat ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata
     3421 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
     3481 gatatatttt tatcttgtgc aatgtaacat cagagatttt gaaggccaaa taggccgttc
     3541 agatccttcc gtatttagcc agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc
     3601 atgagaacga accattgaga tcatgcttac tttgcatgtc actcaaaaat tttgcctcaa
     3661 aactggtgag ctgaattttt gcagttaaag catcgtgtag tgtttttctt agtccgttac
     3721 gtaggtagga atctgatgta atggttgttg gtattttgtc accattcatt tttatctggt
     3781 tgttctcaag ttcggttacg agatccattt gtctatctag ttcaacttgg aaaatcaacg
     3841 tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat attgctgtaa gtgtttaaat
     3901 ctttacttat tggtttcaaa acccattggt taagcctttt aaactcatgg tagttatttt
     3961 caagcattaa catgaactta aattcatcaa ggctaatctc tatatttgcc ttgtgagttt
     4021 tcttttgtgt tagttctttt aataaccact cataaatcct catagagtat ttgttttcaa
     4081 aagacttaac atgttccaga ttatatttta tgaatttttt taactggaaa agataaggca
     4141 atatctcttc actaaaaact aattctaatt tttcgcttga gaacttggca tagtttgtcc
     4201 actggaaaat ccgaaagcct ttaaccaaag gattcctgat ttccacagtt ctcgtcatca
     4261 gctctctggt tgctttagct aatacaccat aagcattttc cctactgatg ttcatcatct
     4321 gagcgtattg gttataagtg aacgataccg tccgttcttt ccttgtaggg ttttcaatcg
     4381 tggggttgag tagtgccaca cagcataaaa ttagcttggt ttcatgctcc gttaagtcat
     4441 agcgactaat cgctagttca tttgctttga aaacaactaa ttcagacata catctcaatt
     4501 ggtctaggtg attttaatca ctataccaat tgagatgggc tagtcaatga taattactag
     4561 tccttttcct ttgagttgtg ggtatctgta aattctgcta gacctttgct ggaaaacttg
     4621 taaattctgc tagaccctct gtaaattccg ctagaccttt gtgtgttttt tttgtttata
     4681 ttcaagtggt tataatttat agaataaaga aagaataaaa aaagataaaa agaatagatc
     4741 ccagccctgt gtataactca ctactttagt cagttccgca gtattacaaa aggatgtcgc
     4801 aaacgctgtt tgctcctcta caaaacagac cttaaaaccc taaaggctta agtagcaccc
     4861 tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc cgaccatcag gcacctgagt
     4921 cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg ctctggcagt gaatgggggt
     4981 aaatggcact acaggcgcct tttatggatt catgcaagga aactacccat aatacaagaa
     5041 aagcccgtca cgggcttctc agggcgtttt atggcgggtc tgctatgtgg tgctatctga
     5101 ctttttgctg ttcagcagtt cctgccctct gattttccag tctgaccact tcggattatc
     5161 ccgtgacagg tcattcagac tggctaatgc acccagtaag gcagcggtat catcaacagg
     5221 cttacccgtc ttactgtcaa gaggacatcc ggtggctgtt ttggcggatg agagaagatt
     5281 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
     5341 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     5401 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcat
//
