LOCUS       sequence-267536-        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     complement(316..333)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      486..572
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      664..691
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             900..1980
                     /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             2044..3024
                     /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      3051..3082
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3098..3913)
                     /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(3098..3913)
                     /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     3821..3840
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3905..3924
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(4024..4974)
                     /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      5022..5244
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gaatgggggt aaatggcact acaggcgcct tttatggatt catgcaagga aactacccat
       61 aatacaagaa aagcccgtca cgggcttctc agggcgtttt atggcgggtc tgctatgtgg
      121 tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag tctgaccact
      181 tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag gcagcggtat
      241 catcaacagg cttacccgtc ttactgtcaa gaggacatcc ggtggctgtt ttggcggatg
      301 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
      361 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
      421 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca
      481 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
      541 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
      601 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
      661 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactcgc tgaggattct
      721 agagcacagc taacaccacg tcgtccctat ctgctgccct aggtctatga gtggttgctg
      781 gataacttta cgggcatgca taaggctcgt ataatatatt cagggagacc acaacggttt
      841 ccctctacaa ataattttgt ttaactttat atctgttatt ttttaaggag gaaaaaaaaa
      901 tgaaatttgg aaactttttg cttacatacc aacctcccca attttcccaa acagaggtaa
      961 tgaaacgttt ggttaaatta ggtcgcatct ctgaggagtg tggttttgat accgtatggt
     1021 tactggagca tcatttcacg gagtttggtt tgcttggtaa cccttatgtc gctgctgcat
     1081 atttacttgg cgcgactaaa aaattgaatg taggaactgc cgctattgtt cttcccacag
     1141 cccatccagt acgccaactt gaagatgtga atttattgga tcaaatgtca aaaggacgat
     1201 ttcggtttgg tatttgccga gggctttaca acaaggactt tcgcgtattc ggaacagata
     1261 tgaataacag tcgcgcctta gcggaatgct ggtacgggct gataaagaat ggcatgacag
     1321 agggatatat ggaagctgat aatgaacata tcaagttcca taaggtaaaa gtaaaccccg
     1381 cggcgtatag cagaggtggc gcaccggttt atgtggtggc tgaatcagct tcgacgactg
     1441 agtgggctgc tcaatttggc ctaccgatga tattaagttg gattataaat actaacgaaa
     1501 agaaagcaca acttgagctt tataatgaag tggctcaaga atatgggcac gatattcata
     1561 atatcgacca ttgcttatca tatataacat ctgtagatca tgactcaatt aaagcgaaag
     1621 agatttgccg gaaatttctg gggcattggt atgattctta tgtgaatgct acgactattt
     1681 ttgatgattc agaccaaaca agaggttatg atttcaataa agggcagtgg cgtgactttg
     1741 tattaaaagg acataaagat actaatcgcc gtattgatta cagttacgaa atcaatcccg
     1801 tgggaacgcc gcaggaatgt attgacataa ttcaaaaaga cattgatgct acaggaatat
     1861 caaatatttg ttgtggattt gaagctaatg gaacagtaga cgaaattatt gcttccatga
     1921 agctcttcca gtctgatgtc atgccatttc ttaaagaaaa acaacgtcgg tctattatat
     1981 tatggcggtg gcggtagcgg cggtggcggt agcggcggtg gcggtagcgg cggtggcggt
     2041 agcaaatttg gattgttctt ccttaacttc atcaattcaa caactgttca agaacagagt
     2101 atagttcgca tgcaggaaat aacggagtat gttgataagt tgaattttga acagatttta
     2161 gtgtatgaaa atcatttttc agataatggt gttgtcggcg ctcctctgac tgtttctggt
     2221 tttctgctcg gtttaacaga gaaaattaaa attggttcat taaatcacat cattacaact
     2281 catcatcctg tccgcatagc ggaggaagct tgcttattgg atcagttaag tgaagggaga
     2341 tttattttag ggtttagtga ttgcgaaaaa aaagatgaaa tgcatttttt taatcgcccg
     2401 gttgaatatc aacagcaact atttgaagag tgttatgaaa tcattaacga tgctttaaca
     2461 acaggctatt gtaatccaga taacgatttt tatagcttcc ctaaaatatc tgtaaatccc
     2521 catgcttata cgccaggcgg acctcggaaa tatgtaacag caaccagtca tcatattgtt
     2581 gagtgggcgg ccaaaaaagg tattcctctc atctttaagt gggatgattc taatgatgtt
     2641 agatatgaat atgctgaaag atataaagcc gttgcggata aatatgacgt tgacctatca
     2701 gagatagacc atcagttaat gatattagtt aactataacg aagatagtaa taaagctaaa
     2761 caagagacgc gtgcatttat tagtgattat gttcttgaaa tgcaccctaa tgaaaatttc
     2821 gaaaataaac ttgaagaaat aattgcagaa aacgctgtcg gaaattatac ggagtgtata
     2881 actgcggcta agttggcaat tgaaaagtgt ggtgcgaaaa gtgtattgct gtcctttgaa
     2941 ccaatgaatg atttgatgag ccaaaaaaat gtaatcaata ttgttgatga taatattaag
     3001 aagtaccaca cggaatatac ctaaacttaa ttaacggcac tcctcagcca agtcaaaagc
     3061 ctccgaccgg aggcttttga ctacatgccc atggcgttta gaaaaactca tcgagcatca
     3121 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     3181 tctgtaatga aggagaaaac tcaccgaggc agttccatag aatggcaaga tcctggtatc
     3241 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     3301 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     3361 gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     3421 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     3481 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     3541 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     3601 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     3661 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     3721 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     3781 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     3841 catataaatc agcatccatg ttggaattta atcgcggcct cgagcaagac gtttcccgtt
     3901 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     3961 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgaagg ccaaataggc
     4021 cgttcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt
     4081 gagccatgag aacgaaccat tgagatcatg cttactttgc atgtcactca aaaattttgc
     4141 ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc
     4201 gttacgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat
     4261 ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat
     4321 caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt
     4381 taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt
     4441 attttcaagc attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg
     4501 agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt
     4561 ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata
     4621 aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt
     4681 tgtccactgg aaaatccgaa agcctttaac caaaggattc ctgatttcca cagttctcgt
     4741 catcagctct ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat
     4801 catctgagcg tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc
     4861 aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa
     4921 gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct
     4981 caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt
     5041 actagtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa
     5101 acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt
     5161 ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat
     5221 agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat
     5281 gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag
     5341 caccctcgca agctcgggca aatcgctgaa tattcctttt gtctccgacc atcaggcacc
     5401 tgagtcgctg tctttttcgt gacattcagt tcgctgcgct cacggctctg gcagt
//
