LOCUS       sequence-263778-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for quadruplet codon CAGG.
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: pED1a20
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      94..180
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      272..299
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             508..1588
                     /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
                     KLFQSDVMPFLKEKQRQAII"
     CDS             1652..2632
                     /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      2659..2690
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2706..3521)
                     /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(2706..3521)
                     /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     3429..3448
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3513..3532
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3632..4582)
                     /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      4630..4852
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(5379..5396)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
       61 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
      121 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
      181 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
      241 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
      301 gcgtttctac aaactcgctg aggattctag agcacagcta acaccacgtc gtccctatct
      361 gctgccctag gtctatgagt ggttgctgga taactttacg ggcatgcata aggctcgtat
      421 aatatattca gggagaccac aacggtttcc ctctacaaat aattttgttt aactttatat
      481 ctgttatttt ttaaggagga aaaaaaaatg aaatttggaa actttttgct tacataccaa
      541 cctccccaat tttcccaaac agaggtaatg aaacgtttgg ttaaattagg tcgcatctct
      601 gaggagtgtg gttttgatac cgtatggtta ctggagcatc atttcacgga gtttggtttg
      661 cttggtaacc cttatgtcgc tgctgcatat ttacttggcg cgactaaaaa attgaatgta
      721 ggaactgccg ctattgttct tcccacagcc catccagtac gccaacttga agatgtgaat
      781 ttattggatc aaatgtcaaa aggacgattt cggtttggta tttgccgagg gctttacaac
      841 aaggactttc gcgtattcgg aacagatatg aataacagtc gcgccttagc ggaatgctgg
      901 tacgggctga taaagaatgg catgacagag ggatatatgg aagctgataa tgaacatatc
      961 aagttccata aggtaaaagt aaaccccgcg gcgtatagca gaggtggcgc accggtttat
     1021 gtggtggctg aatcagcttc gacgactgag tgggctgctc aatttggcct accgatgata
     1081 ttaagttgga ttataaatac taacgaaaag aaagcacaac ttgagcttta taatgaagtg
     1141 gctcaagaat atgggcacga tattcataat atcgaccatt gcttatcata tataacatct
     1201 gtagatcatg actcaattaa agcgaaagag atttgccgga aatttctggg gcattggtat
     1261 gattcttatg tgaatgctac gactattttt gatgattcag accaaacaag aggttatgat
     1321 ttcaataaag ggcagtggcg tgactttgta ttaaaaggac ataaagatac taatcgccgt
     1381 attgattaca gttacgaaat caatcccgtg ggaacgccgc aggaatgtat tgacataatt
     1441 caaaaagaca ttgatgctac aggaatatca aatatttgtt gtggatttga agctaatgga
     1501 acagtagacg aaattattgc ttccatgaag ctcttccagt ctgatgtcat gccatttctt
     1561 aaagaaaaac aacgtcaggc tattatatta tggcggtggc ggtagcggcg gtggcggtag
     1621 cggcggtggc ggtagcggcg gtggcggtag caaatttgga ttgttcttcc ttaacttcat
     1681 caattcaaca actgttcaag aacagagtat agttcgcatg caggaaataa cggagtatgt
     1741 tgataagttg aattttgaac agattttagt gtatgaaaat catttttcag ataatggtgt
     1801 tgtcggcgct cctctgactg tttctggttt tctgctcggt ttaacagaga aaattaaaat
     1861 tggttcatta aatcacatca ttacaactca tcatcctgtc cgcatagcgg aggaagcttg
     1921 cttattggat cagttaagtg aagggagatt tattttaggg tttagtgatt gcgaaaaaaa
     1981 agatgaaatg cattttttta atcgcccggt tgaatatcaa cagcaactat ttgaagagtg
     2041 ttatgaaatc attaacgatg ctttaacaac aggctattgt aatccagata acgattttta
     2101 tagcttccct aaaatatctg taaatcccca tgcttatacg ccaggcggac ctcggaaata
     2161 tgtaacagca accagtcatc atattgttga gtgggcggcc aaaaaaggta ttcctctcat
     2221 ctttaagtgg gatgattcta atgatgttag atatgaatat gctgaaagat ataaagccgt
     2281 tgcggataaa tatgacgttg acctatcaga gatagaccat cagttaatga tattagttaa
     2341 ctataacgaa gatagtaata aagctaaaca agagacgcgt gcatttatta gtgattatgt
     2401 tcttgaaatg caccctaatg aaaatttcga aaataaactt gaagaaataa ttgcagaaaa
     2461 cgctgtcgga aattatacgg agtgtataac tgcggctaag ttggcaattg aaaagtgtgg
     2521 tgcgaaaagt gtattgctgt cctttgaacc aatgaatgat ttgatgagcc aaaaaaatgt
     2581 aatcaatatt gttgatgata atattaagaa gtaccacacg gaatatacct aaacttaatt
     2641 aacggcactc ctcagccaag tcaaaagcct ccgaccggag gcttttgact acatgcccat
     2701 ggcgtttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     2761 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     2821 ttccatagaa tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     2881 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     2941 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     3001 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     3061 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     3121 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     3181 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     3241 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     3301 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     3361 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     3421 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     3481 cgcggcctcg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     3541 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     3601 catcagagat tttgaaggcc aaataggccg ttcagatcct tccgtattta gccagtatgt
     3661 tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatgct
     3721 tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta
     3781 aagcatcgtg tagtgttttt cttagtccgt tacgtaggta ggaatctgat gtaatggttg
     3841 ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca
     3901 tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa
     3961 ccaccaattt catattgctg taagtgttta aatctttact tattggtttc aaaacccatt
     4021 ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat
     4081 caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc
     4141 actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt
     4201 ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta
     4261 atttttcgct tgagaacttg gcatagtttg tccactggaa aatccgaaag cctttaacca
     4321 aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac
     4381 cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata
     4441 ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata
     4501 aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt
     4561 tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa tcactatacc
     4621 aattgagatg ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct
     4681 gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt
     4741 ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa
     4801 agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt
     4861 agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca
     4921 gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata
     4981 ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc
     5041 gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg
     5101 attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt
     5161 tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc
     5221 tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa
     5281 tgcacccagt aaggcagcgg tatcatcaac aggcttaccc gtcttactgt caagaggaca
     5341 tccggtggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     5401 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcc
//
