LOCUS       sequence-259099-        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"
     CDS             complement(1..951)
                     /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      999..1221
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(1748..1765)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1918..2004
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2096..2123
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2332..3412
                     /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             3476..4456
                     /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      4483..4514
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4530..5345)
                     /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(4530..5345)
                     /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     5253..5272
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5337..5356
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
       61 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
      121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
      181 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
      241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
      301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
      361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
      421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
      481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
      541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
      601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
      661 ccactggaaa atccgaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
      721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
      781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
      841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
      901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
      961 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     1021 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     1141 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     1561 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     1681 ggcttacccg tcttactgtc aagaggacat ccggtggctg ttttggcgga tgagagaaga
     1741 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     1801 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     1861 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     1921 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1981 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     2041 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     2101 gccatcctga cggatggcct ttttgcgttt ctacaaactc gctgaggatt ctagagcaca
     2161 gctaacacca cgtcgtccct atctgctgcc ctaggtctat gagtggttgc tggataactt
     2221 tacgggcatg cataaggctc gtataatata ttcagggaga ccacaacggt ttccctctac
     2281 aaataatttt gtttaacttt atatctgtta ttttttaagg aggaaaaaaa aatgaaattt
     2341 ggaaactttt tgcttacata ccaacctccc caattttccc aaacagaggt aatgaaacgt
     2401 ttggttaaat taggtcgcat ctctgaggag tgtggttttg ataccgtatg gttactggag
     2461 catcatttca cggagtttgg tttgcttggt aacccttatg tcgctgctgc atatttactt
     2521 ggcgcgacta aaaaattgaa tgtaggaact gccgctattg ttcttcccac agcccatcca
     2581 gtacgccaac ttgaagatgt gaatttattg gatcaaatgt caaaaggacg atttcggttt
     2641 ggtatttgcc gagggcttta caacaaggac tttcgcgtat tcggaacaga tatgaataac
     2701 agtcgcgcct tagcggaatg ctggtacggg ctgataaaga atggcatgac agagggatat
     2761 atggaagctg ataatgaaca tatcaagttc cataaggtaa aagtaaaccc cgcggcgtat
     2821 agcagaggtg gcgcaccggt ttatgtggtg gctgaatcag cttcgacgac tgagtgggct
     2881 gctcaatttg gcctaccgat gatattaagt tggattataa atactaacga aaagaaagca
     2941 caacttgagc tttataatga agtggctcaa gaatatgggc acgatattca taatatcgac
     3001 cattgcttat catatataac atctgtagat catgactcaa ttaaagcgaa agagatttgc
     3061 cggaaatttc tggggcattg gtatgattct tatgtgaatg ctacgactat ttttgatgat
     3121 tcagaccaaa caagaggtta tgatttcaat aaagggcagt ggcgtgactt tgtattaaaa
     3181 ggacataaag atactaatcg ccgtattgat tacagttacg aaatcaatcc cgtgggaacg
     3241 ccgcaggaat gtattgacat aattcaaaaa gacattgatg ctacaggaat atcaaatatt
     3301 tgttgtggat ttgaagctaa tggaacagta gacgaaatta ttgcttccat gaagctcttc
     3361 cagtctgatg tcatgccatt tcttaaagaa aaacaacgtc aggctattat attatggcgg
     3421 tggcggtagc ggcggtggcg gtagcggcgg tggcggtagc ggcggtggcg gtagcaaatt
     3481 tggattgttc ttccttaact tcatcaattc aacaactgtt caagaacaga gtatagttcg
     3541 catgcaggaa ataacggagt atgttgataa gttgaatttt gaacagattt tagtgtatga
     3601 aaatcatttt tcagataatg gtgttgtcgg cgctcctctg actgtttctg gttttctgct
     3661 cggtttaaca gagaaaatta aaattggttc attaaatcac atcattacaa ctcatcatcc
     3721 tgtccgcata gcggaggaag cttgcttatt ggatcagtta agtgaaggga gatttatttt
     3781 agggtttagt gattgcgaaa aaaaagatga aatgcatttt tttaatcgcc cggttgaata
     3841 tcaacagcaa ctatttgaag agtgttatga aatcattaac gatgctttaa caacaggcta
     3901 ttgtaatcca gataacgatt tttatagctt ccctaaaata tctgtaaatc cccatgctta
     3961 tacgccaggc ggacctcgga aatatgtaac agcaaccagt catcatattg ttgagtgggc
     4021 ggccaaaaaa ggtattcctc tcatctttaa gtgggatgat tctaatgatg ttagatatga
     4081 atatgctgaa agatataaag ccgttgcgga taaatatgac gttgacctat cagagataga
     4141 ccatcagtta atgatattag ttaactataa cgaagatagt aataaagcta aacaagagac
     4201 gcgtgcattt attagtgatt atgttcttga aatgcaccct aatgaaaatt tcgaaaataa
     4261 acttgaagaa ataattgcag aaaacgctgt cggaaattat acggagtgta taactgcggc
     4321 taagttggca attgaaaagt gtggtgcgaa aagtgtattg ctgtcctttg aaccaatgaa
     4381 tgatttgatg agccaaaaaa atgtaatcaa tattgttgat gataatatta agaagtacca
     4441 cacggaatat acctaaactt aattaacggc actcctcagc caagtcaaaa gcctccgacc
     4501 ggaggctttt gactacatgc ccatggcgtt tagaaaaact catcgagcat caaatgaaac
     4561 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     4621 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     4681 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     4741 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     4801 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     4861 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     4921 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     4981 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     5041 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     5101 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     5161 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     5221 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     5281 tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg
     5341 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     5401 atatttttat cttgtgcaat gtaacatcag agattttgaa ggccaaatag gccgt
//
