LOCUS       sequence-350043-        6504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual luciferase reporter for SARS-CoV-2 programmed -1 ribosomal 
            frameshifting. In-frame stop codon inserted after FLuc (negative 
            control)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6504)
  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 6504)
  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-FLucStop
FEATURES             Location/Qualifiers
     source          1..6504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     174..193
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      282..291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             288..1940
                     /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             2039..2968
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="ASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             3020..3811
                     /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(3071..3090)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3681..3700
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3875..3923
                     /note="synthetic polyadenylation signal"
     promoter        4079..4183
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4184..4252,4253..5044)
                     /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(4402..4421)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5202..5790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5282..5301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5906..6189
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        join(6246..6504,1..241)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6281..6304)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
       61 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
      121 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
      181 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
      241 gacctgcagc ccaagcttgg caatccggta ctgttggtaa agccaccatg gaagatgcca
      301 aaaacattaa gaagggccca gcgccattct acccactcga agacgggacc gccggcgagc
      361 agctgcacaa agccatgaag cgctacgccc tggtgcccgg caccatcgcc tttaccgacg
      421 cacatatcga ggtggacatt acctacgccg agtacttcga gatgagcgtt cggctggcag
      481 aagctatgaa gcgctatggg ctgaatacaa accatcggat cgtggtgtgc agcgagaata
      541 gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat cggtgtggct gtggccccag
      601 ctaacgacat ctacaacgag cgcgagctgc tgaacagcat gggcatcagc cagcccaccg
      661 tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga
      721 tcatacaaaa gatcatcatc atggatagca agaccgacta ccagggcttc caaagcatgt
      781 acaccttcgt gacttcccat ttgccacccg gcttcaacga gtacgacttc gtgcccgaga
      841 gcttcgaccg ggacaaaacc atcgccctga tcatgaacag tagtggcagt accggattgc
      901 ccaagggcgt agccctaccg caccgcaccg cttgtgtccg attcagtcat gcccgcgacc
      961 ccatcttcgg caaccagatc atccccgaca ccgctatcct cagcgtggtg ccatttcacc
     1021 acggcttcgg catgttcacc acgctgggct acttgatctg cggctttcgg gtcgtgctca
     1081 tgtaccgctt cgaggaggag ctattcttgc gcagcttgca agactataag attcaatctg
     1141 ccctgctggt gcccacacta tttagcttct tcgctaagag cactctcatc gacaagtacg
     1201 acctaagcaa cttgcacgag atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg
     1261 aggccgtggc caaacgcttc cacctaccag gcatccgcca gggctacggc ctgacagaaa
     1321 caaccagcgc cattctgatc acccccgaag gggacgacaa gcctggcgca gtaggcaagg
     1381 tggtgccctt cttcgaggct aaggtggtgg acttggacac cggtaagaca ctgggtgtga
     1441 accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat gagcggctac gttaacaacc
     1501 ccgaggctac aaacgctctc atcgacaagg acggctggct gcacagcggc gacatcgcct
     1561 actgggacga ggacgagcac ttcttcatcg tggaccggct gaagagcctg atcaaataca
     1621 agggctacca ggtagcccca gccgaactgg agagcatcct gctgcaacac cccaacatct
     1681 tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg
     1741 tcgtgctgga acacggtaaa accatgaccg agaaggagat cgtggactat gtggccagcc
     1801 aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag
     1861 gactgaccgg caagttggac gcccgcaaga tccgcgagat tctcattaag gccaagaagg
     1921 gcggcaagat cgccgtgtaa aataactcgt ttttaaacgg gtttgcggtg taagtgcagc
     1981 ccgtcttaca ccgtgcggca caggcactag tactgatgtc gtatacaggg cttttgacgc
     2041 ttccaaggtg tacgaccccg agcaacgcaa acgcatgatc actgggcctc agtggtgggc
     2101 tcgctgcaag caaatgaacg tgctggactc cttcatcaac tactatgatt ccgagaagca
     2161 cgccgagaac gccgtgattt ttctgcatgg taacgctgcc tccagctacc tgtggaggca
     2221 cgtcgtgcct cacatcgagc ccgtggctag atgcatcatc cctgatctga tcggaatggg
     2281 taagtccggc aagagcggga atggctcata tcgcctcctg gatcactaca agtacctcac
     2341 cgcttggttc gagctgctga accttccaaa gaaaatcatc tttgtgggcc acgactgggg
     2401 ggcttgtctg gcctttcact actcctacga gcaccaagac aagatcaagg ccatcgtcca
     2461 tgctgagagt gtcgtggacg tgatcgagtc ctgggacgag tggcctgaca tcgaggagga
     2521 tatcgccctg atcaagagcg aagagggcga gaaaatggtg cttgagaata acttcttcgt
     2581 cgagaccatg ctcccaagca agatcatgcg gaaactggag cctgaggagt tcgctgccta
     2641 cctggagcca ttcaaggaga agggcgaggt tagacggcct accctctcct ggcctcgcga
     2701 gatccctctc gttaagggag gcaagcccga cgtcgtccag attgtccgca actacaacgc
     2761 ctaccttcgg gccagcgacg atctgcctaa gatgttcatc gagtccgacc ctgggttctt
     2821 ttccaacgct attgtcgagg gagctaagaa gttccctaac accgagttcg tgaaggtgaa
     2881 gggcctccac ttcagccagg aggacgctcc agatgaaatg ggtaagtaca tcaagagctt
     2941 cgtggagcgc gtgctgaaga acgagcagac cggtggtggg agcggaggtg gcggatcagg
     3001 tggcggaggc tccggaggga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3061 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3121 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3181 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3241 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3301 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3361 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3421 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3481 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3541 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3601 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3661 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3721 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3781 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3841 caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     3901 tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg atcctctttg cgcttgcgtt
     3961 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     4021 actggctttc tacgtaatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4081 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4141 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4201 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4261 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4321 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4381 gatgagcact ttcaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4441 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4501 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4561 catgagtgat aacactgcgg ccaacttact tctgacaact atcggaggac cgaaggagct
     4621 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4681 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4741 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4801 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4861 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4921 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4981 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5041 gtaattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta
     5101 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5161 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5221 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5281 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5341 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5401 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5461 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5521 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5581 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5641 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5701 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5761 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     5821 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5881 atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg atggcccttc
     5941 gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg
     6001 cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc gcgattgcgg
     6061 cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc gcgctgaacg
     6121 cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt
     6181 agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat
     6241 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
     6301 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
     6361 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
     6421 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
     6481 agcacgtctc actagtctcg tgca
//
