LOCUS       sequence-263781-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for quadruplet codon AGGG.
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: pED1a34
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      117..203
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      295..322
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             531..1611
                     /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
                     KLFQSDVMPFLKEKQRRAII"
     CDS             1675..2655
                     /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      2682..2713
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2729..3544)
                     /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(2729..3544)
                     /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     3452..3471
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3536..3555
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3655..4605)
                     /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      4653..4875
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(5402..5419)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
       61 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
      121 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
      181 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
      241 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
      301 ccatcctgac ggatggcctt tttgcgtttc tacaaactcg ctgaggattc tagagcacag
      361 ctaacaccac gtcgtcccta tctgctgccc taggtctatg agtggttgct ggataacttt
      421 acgggcatgc ataaggctcg tataatatat tcagggagac cacaacggtt tccctctaca
      481 aataattttg tttaacttta tatctgttat tttttaagga ggaaaaaaaa atgaaatttg
      541 gaaacttttt gcttacatac caacctcccc aattttccca aacagaggta atgaaacgtt
      601 tggttaaatt aggtcgcatc tctgaggagt gtggttttga taccgtatgg ttactggagc
      661 atcatttcac ggagtttggt ttgcttggta acccttatgt cgctgctgca tatttacttg
      721 gcgcgactaa aaaattgaat gtaggaactg ccgctattgt tcttcccaca gcccatccag
      781 tacgccaact tgaagatgtg aatttattgg atcaaatgtc aaaaggacga tttcggtttg
      841 gtatttgccg agggctttac aacaaggact ttcgcgtatt cggaacagat atgaataaca
      901 gtcgcgcctt agcggaatgc tggtacgggc tgataaagaa tggcatgaca gagggatata
      961 tggaagctga taatgaacat atcaagttcc ataaggtaaa agtaaacccc gcggcgtata
     1021 gcagaggtgg cgcaccggtt tatgtggtgg ctgaatcagc ttcgacgact gagtgggctg
     1081 ctcaatttgg cctaccgatg atattaagtt ggattataaa tactaacgaa aagaaagcac
     1141 aacttgagct ttataatgaa gtggctcaag aatatgggca cgatattcat aatatcgacc
     1201 attgcttatc atatataaca tctgtagatc atgactcaat taaagcgaaa gagatttgcc
     1261 ggaaatttct ggggcattgg tatgattctt atgtgaatgc tacgactatt tttgatgatt
     1321 cagaccaaac aagaggttat gatttcaata aagggcagtg gcgtgacttt gtattaaaag
     1381 gacataaaga tactaatcgc cgtattgatt acagttacga aatcaatccc gtgggaacgc
     1441 cgcaggaatg tattgacata attcaaaaag acattgatgc tacaggaata tcaaatattt
     1501 gttgtggatt tgaagctaat ggaacagtag acgaaattat tgcttccatg aagctcttcc
     1561 agtctgatgt catgccattt cttaaagaaa aacaacgtag ggctattata ttatggcggt
     1621 ggcggtagcg gcggtggcgg tagcggcggt ggcggtagcg gcggtggcgg tagcaaattt
     1681 ggattgttct tccttaactt catcaattca acaactgttc aagaacagag tatagttcgc
     1741 atgcaggaaa taacggagta tgttgataag ttgaattttg aacagatttt agtgtatgaa
     1801 aatcattttt cagataatgg tgttgtcggc gctcctctga ctgtttctgg ttttctgctc
     1861 ggtttaacag agaaaattaa aattggttca ttaaatcaca tcattacaac tcatcatcct
     1921 gtccgcatag cggaggaagc ttgcttattg gatcagttaa gtgaagggag atttatttta
     1981 gggtttagtg attgcgaaaa aaaagatgaa atgcattttt ttaatcgccc ggttgaatat
     2041 caacagcaac tatttgaaga gtgttatgaa atcattaacg atgctttaac aacaggctat
     2101 tgtaatccag ataacgattt ttatagcttc cctaaaatat ctgtaaatcc ccatgcttat
     2161 acgccaggcg gacctcggaa atatgtaaca gcaaccagtc atcatattgt tgagtgggcg
     2221 gccaaaaaag gtattcctct catctttaag tgggatgatt ctaatgatgt tagatatgaa
     2281 tatgctgaaa gatataaagc cgttgcggat aaatatgacg ttgacctatc agagatagac
     2341 catcagttaa tgatattagt taactataac gaagatagta ataaagctaa acaagagacg
     2401 cgtgcattta ttagtgatta tgttcttgaa atgcacccta atgaaaattt cgaaaataaa
     2461 cttgaagaaa taattgcaga aaacgctgtc ggaaattata cggagtgtat aactgcggct
     2521 aagttggcaa ttgaaaagtg tggtgcgaaa agtgtattgc tgtcctttga accaatgaat
     2581 gatttgatga gccaaaaaaa tgtaatcaat attgttgatg ataatattaa gaagtaccac
     2641 acggaatata cctaaactta attaacggca ctcctcagcc aagtcaaaag cctccgaccg
     2701 gaggcttttg actacatgcc catggcgttt agaaaaactc atcgagcatc aaatgaaact
     2761 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     2821 aaggagaaaa ctcaccgagg cagttccata gaatggcaag atcctggtat cggtctgcga
     2881 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     2941 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     3001 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     3061 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     3121 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     3181 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     3241 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     3301 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     3361 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     3421 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     3481 cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc
     3541 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     3601 tatttttatc ttgtgcaatg taacatcaga gattttgaag gccaaatagg ccgttcagat
     3661 ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga
     3721 gaacgaacca ttgagatcat gcttactttg catgtcactc aaaaattttg cctcaaaact
     3781 ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttacgtag
     3841 gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt
     3901 ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc
     3961 agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt
     4021 acttattggt ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag
     4081 cattaacatg aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt
     4141 ttgtgttagt tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga
     4201 cttaacatgt tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat
     4261 ctcttcacta aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg
     4321 gaaaatccga aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc
     4381 tctggttgct ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc
     4441 gtattggtta taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg
     4501 gttgagtagt gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg
     4561 actaatcgct agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc
     4621 taggtgattt taatcactat accaattgag atgggctagt caatgataat tactagtcct
     4681 tttcctttga gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa
     4741 ttctgctaga ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca
     4801 agtggttata atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag
     4861 ccctgtgtat aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac
     4921 gctgtttgct cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc
     4981 aagctcgggc aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct
     5041 gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat
     5101 ggcactacag gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc
     5161 ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt
     5221 ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt
     5281 gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta
     5341 cccgtcttac tgtcaagagg acatccggtg gctgttttgg cggatgagag aagattttca
     5401 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcc
//
