LOCUS       sequence-350039-        6501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual luciferase reporter for SARS-CoV-2 programmed -1 ribosomal 
            frameshifting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6501)
  AUTHORS   Sun Y, Abriola L, Niederer RO, Pedersen SF, Alfajaro MM, Silva 
            Monteiro V, Wilen CB, Ho YC, Gilbert WV, Surovtseva YV, Lindenbach 
            BD, Guo JU
  TITLE     Restriction of SARS-CoV-2 replication by targeting programmed -1 
            ribosomal frameshifting.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Jun 29;118(26). pii: 2023051118. doi:
            10.1073/pnas.2023051118.
  PUBMED    34185680
REFERENCE   2  (bases 1 to 6501)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-06-29"; volume: "118(26). pii: 
            2023051118. doi"; pages: " 10.1073/pnas.2023051118"
COMMENT     Sequence Label: pFRLuc-CoV2-WT
FEATURES             Location/Qualifiers
     source          1..6501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..111
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      200..209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             206..1855
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             1954..2883
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="ASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             2935..3726
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2986..3005)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3596..3615
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3790..3838
                     /note="synthetic polyadenylation signal"
     promoter        3994..4098
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4099..4167,4168..4959)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4317..4336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5117..5705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5197..5216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5821..6104
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        join(6161..6501,1..159)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6196..6219)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
       61 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
      121 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttggca
      181 atccggtact gttggtaaag ccaccatgga agatgccaaa aacattaaga agggcccagc
      241 gccattctac ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg
      301 ctacgccctg gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac
      361 ctacgccgag tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct
      421 gaatacaaac catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt
      481 gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg
      541 cgagctgctg aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg
      601 gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat
      661 ggatagcaag accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt
      721 gccacccggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat
      781 cgccctgatc atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca
      841 ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat
      901 ccccgacacc gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac
      961 gctgggctac ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct
     1021 attcttgcgc agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt
     1081 tagcttcttc gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat
     1141 cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca
     1201 cctaccaggc atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac
     1261 ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa
     1321 ggtggtggac ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt
     1381 ccgtggcccc atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat
     1441 cgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt
     1501 cttcatcgtg gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc
     1561 cgaactggag agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct
     1621 gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac
     1681 catgaccgag aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct
     1741 gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc
     1801 ccgcaagatc cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgaataa
     1861 ctcgttttta aacgggtttg cggtgtaagt gcagcccgtc ttacaccgtg cggcacaggc
     1921 actagtactg atgtcgtata cagggctttt gacgcttcca aggtgtacga ccccgagcaa
     1981 cgcaaacgca tgatcactgg gcctcagtgg tgggctcgct gcaagcaaat gaacgtgctg
     2041 gactccttca tcaactacta tgattccgag aagcacgccg agaacgccgt gatttttctg
     2101 catggtaacg ctgcctccag ctacctgtgg aggcacgtcg tgcctcacat cgagcccgtg
     2161 gctagatgca tcatccctga tctgatcgga atgggtaagt ccggcaagag cgggaatggc
     2221 tcatatcgcc tcctggatca ctacaagtac ctcaccgctt ggttcgagct gctgaacctt
     2281 ccaaagaaaa tcatctttgt gggccacgac tggggggctt gtctggcctt tcactactcc
     2341 tacgagcacc aagacaagat caaggccatc gtccatgctg agagtgtcgt ggacgtgatc
     2401 gagtcctggg acgagtggcc tgacatcgag gaggatatcg ccctgatcaa gagcgaagag
     2461 ggcgagaaaa tggtgcttga gaataacttc ttcgtcgaga ccatgctccc aagcaagatc
     2521 atgcggaaac tggagcctga ggagttcgct gcctacctgg agccattcaa ggagaagggc
     2581 gaggttagac ggcctaccct ctcctggcct cgcgagatcc ctctcgttaa gggaggcaag
     2641 cccgacgtcg tccagattgt ccgcaactac aacgcctacc ttcgggccag cgacgatctg
     2701 cctaagatgt tcatcgagtc cgaccctggg ttcttttcca acgctattgt cgagggagct
     2761 aagaagttcc ctaacaccga gttcgtgaag gtgaagggcc tccacttcag ccaggaggac
     2821 gctccagatg aaatgggtaa gtacatcaag agcttcgtgg agcgcgtgct gaagaacgag
     2881 cagaccggtg gtgggagcgg aggtggcgga tcaggtggcg gaggctccgg agggattgaa
     2941 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3001 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3061 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3121 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3181 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3241 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3301 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3361 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3421 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3481 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3541 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3601 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3661 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3721 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3781 gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     3841 tcgatagcga taaggatcct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     3901 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt aatggtttct
     3961 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4021 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4081 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4141 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4201 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4261 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcactttcaa agttctgcta
     4321 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4381 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4441 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4501 ttacttctga caactatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4561 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4621 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4681 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4741 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4801 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4861 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4921 atcgctgaga taggtgcctc actgattaag cattggtaat tcgaaatgac cgaccaagcg
     4981 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5041 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5101 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5161 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5221 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5281 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5341 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5401 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5461 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5521 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5581 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5641 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5701 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5761 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     5821 ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa
     5881 tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta
     5941 ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca
     6001 tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag
     6061 ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact
     6121 ccggccacga tgcgtccggc gtagaggatc gagatctacc gggtagggga ggcgcttttc
     6181 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     6241 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     6301 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     6361 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     6421 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     6481 ctttgctcct tcgctttctg g
//
