LOCUS       sequence-259097-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for quadruplet codon TAGA.
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: pED1a6
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     293..312
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     377..396
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(496..1446)
                     /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      1494..1716
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(2243..2260)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2413..2499
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2591..2618
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2827..3907
                     /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
                     KLFQSDVMPFLKEKQR*TII"
     CDS             3971..4951
                     /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      4978..5009
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(join(5025..5455,1..385))
                     /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(5025..5455,1..385))
                     /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 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
       61 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
      121 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
      181 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
      241 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
      301 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag
      361 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
      421 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgaa
      481 ggccaaatag gccgttcaga tccttccgta tttagccagt atgttctcta gtgtggttcg
      541 ttgtttttgc gtgagccatg agaacgaacc attgagatca tgcttacttt gcatgtcact
      601 caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt
      661 ttttcttagt ccgttacgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc
      721 attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc
      781 aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt
      841 gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa
      901 ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat
      961 atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat
     1021 agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa
     1081 ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa
     1141 cttggcatag tttgtccact ggaaaatccg aaagccttta accaaaggat tcctgatttc
     1201 cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct
     1261 actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc gttctttcct
     1321 tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc
     1381 atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc
     1441 agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag
     1501 tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat tctgctagac
     1561 ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg
     1621 tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa
     1681 gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta
     1741 ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa
     1801 aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga
     1861 ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc
     1921 tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac
     1981 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc
     2041 tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct
     2101 gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca
     2161 gcggtatcat caacaggctt acccgtctta ctgtcaagag gacatccggt ggctgttttg
     2221 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     2281 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     2341 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     2401 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2461 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     2521 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     2581 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcgctga
     2641 ggattctaga gcacagctaa caccacgtcg tccctatctg ctgccctagg tctatgagtg
     2701 gttgctggat aactttacgg gcatgcataa ggctcgtata atatattcag ggagaccaca
     2761 acggtttccc tctacaaata attttgttta actttatatc tgttattttt taaggaggaa
     2821 aaaaaaatga aatttggaaa ctttttgctt acataccaac ctccccaatt ttcccaaaca
     2881 gaggtaatga aacgtttggt taaattaggt cgcatctctg aggagtgtgg ttttgatacc
     2941 gtatggttac tggagcatca tttcacggag tttggtttgc ttggtaaccc ttatgtcgct
     3001 gctgcatatt tacttggcgc gactaaaaaa ttgaatgtag gaactgccgc tattgttctt
     3061 cccacagccc atccagtacg ccaacttgaa gatgtgaatt tattggatca aatgtcaaaa
     3121 ggacgatttc ggtttggtat ttgccgaggg ctttacaaca aggactttcg cgtattcgga
     3181 acagatatga ataacagtcg cgccttagcg gaatgctggt acgggctgat aaagaatggc
     3241 atgacagagg gatatatgga agctgataat gaacatatca agttccataa ggtaaaagta
     3301 aaccccgcgg cgtatagcag aggtggcgca ccggtttatg tggtggctga atcagcttcg
     3361 acgactgagt gggctgctca atttggccta ccgatgatat taagttggat tataaatact
     3421 aacgaaaaga aagcacaact tgagctttat aatgaagtgg ctcaagaata tgggcacgat
     3481 attcataata tcgaccattg cttatcatat ataacatctg tagatcatga ctcaattaaa
     3541 gcgaaagaga tttgccggaa atttctgggg cattggtatg attcttatgt gaatgctacg
     3601 actatttttg atgattcaga ccaaacaaga ggttatgatt tcaataaagg gcagtggcgt
     3661 gactttgtat taaaaggaca taaagatact aatcgccgta ttgattacag ttacgaaatc
     3721 aatcccgtgg gaacgccgca ggaatgtatt gacataattc aaaaagacat tgatgctaca
     3781 ggaatatcaa atatttgttg tggatttgaa gctaatggaa cagtagacga aattattgct
     3841 tccatgaagc tcttccagtc tgatgtcatg ccatttctta aagaaaaaca acgttagact
     3901 attatattat ggcggtggcg gtagcggcgg tggcggtagc ggcggtggcg gtagcggcgg
     3961 tggcggtagc aaatttggat tgttcttcct taacttcatc aattcaacaa ctgttcaaga
     4021 acagagtata gttcgcatgc aggaaataac ggagtatgtt gataagttga attttgaaca
     4081 gattttagtg tatgaaaatc atttttcaga taatggtgtt gtcggcgctc ctctgactgt
     4141 ttctggtttt ctgctcggtt taacagagaa aattaaaatt ggttcattaa atcacatcat
     4201 tacaactcat catcctgtcc gcatagcgga ggaagcttgc ttattggatc agttaagtga
     4261 agggagattt attttagggt ttagtgattg cgaaaaaaaa gatgaaatgc atttttttaa
     4321 tcgcccggtt gaatatcaac agcaactatt tgaagagtgt tatgaaatca ttaacgatgc
     4381 tttaacaaca ggctattgta atccagataa cgatttttat agcttcccta aaatatctgt
     4441 aaatccccat gcttatacgc caggcggacc tcggaaatat gtaacagcaa ccagtcatca
     4501 tattgttgag tgggcggcca aaaaaggtat tcctctcatc tttaagtggg atgattctaa
     4561 tgatgttaga tatgaatatg ctgaaagata taaagccgtt gcggataaat atgacgttga
     4621 cctatcagag atagaccatc agttaatgat attagttaac tataacgaag atagtaataa
     4681 agctaaacaa gagacgcgtg catttattag tgattatgtt cttgaaatgc accctaatga
     4741 aaatttcgaa aataaacttg aagaaataat tgcagaaaac gctgtcggaa attatacgga
     4801 gtgtataact gcggctaagt tggcaattga aaagtgtggt gcgaaaagtg tattgctgtc
     4861 ctttgaacca atgaatgatt tgatgagcca aaaaaatgta atcaatattg ttgatgataa
     4921 tattaagaag taccacacgg aatataccta aacttaatta acggcactcc tcagccaagt
     4981 caaaagcctc cgaccggagg cttttgacta catgcccatg gcgtttagaa aaactcatcg
     5041 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     5101 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     5161 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     5221 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     5281 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     5341 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     5401 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggc
//
