LOCUS       sequence-350040-        6492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual luciferase reporter for SARS-CoV-2 programmed -1 ribosomal 
            frameshifting. Slippery sequence (SS) deleted (negative control)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6492)
  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 6492)
  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-delSS
FEATURES             Location/Qualifiers
     source          1..6492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(39..538)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(87..106)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     480..503
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    595..878
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      994..1582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1483..1502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1740..2531,2532..2600))
                     /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     2363..2382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2601..2705)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2861..2909
                     /note="synthetic polyadenylation signal"
     CDS             complement(2973..3764)
                     /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(3084..3103)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3694..3713
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3816..4745)
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="ASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             complement(4835..6484)
                     /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"
     regulatory      6481..6490
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 taccaacagt accggattgc caagcttggg ctgcaggtcg aaaggcccgg agatgaggaa
       61 gaggagaaca gcgcggcaga cgtgcgcttt tgaagcgtgc agaatgccgg gcctccggag
      121 gaccttcggg cgcccgcccc gcccctgagc ccgcccctga gcccgccccc ggacccaccc
      181 cttcccagcc tctgagccca gaaagcgaag gagcaaagct gctattggcc gctgccccaa
      241 aggcctaccc gcttccattg ctcagcggtg ctgtccatct gcacgagact agtgagacgt
      301 gctacttcca tttgtcacgt cctgcacgac gcgagctgcg gggcgggggg gaacttcctg
      361 actaggggag gagtagaagg tggcgcgaag gggccaccaa agaacggagc cggttggcgc
      421 ctaccggtgg atgtggaatg tgtgcgaggc cagaggccac ttgtgtagcg ccaagtgccc
      481 agcggggctg ctaaagcgca tgctccagac tgccttggga aaagcgcctc ccctacccgg
      541 tagatctcga tcctctacgc cggacgcatc gtggccggag tactgcggtt tatctcgatg
      601 gctacgaggg cagacagtaa gtggatttac cataatccct taattgtacg caccgctaaa
      661 acgcgttcag cgcgatcacg gcagcagaca ggtaaaaatg gcaacaaacc acccgaaaaa
      721 ctgccgcaat cgcgcctgat aaattttaac cgtatgaata cctatgcaac cagagggtac
      781 aggccacatt acccccactt aatccactga agctgccatt tttcatggtt tcaccatccc
      841 agcgaagggc catccagcgt gcgttcctgt atttccggac tataaaagga tctaggtgaa
      901 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
      961 gtcagacccc gtagaaaaga tcaaaggatc ttcttgaaat cctttttttc tgcgcgtaat
     1021 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1081 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1141 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1201 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1261 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1321 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1381 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1441 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1501 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1561 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1621 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     1681 tattaccgcc tttgagtgag ctgataccgg ttgggcgtcg cttggtcggt catttcgaat
     1741 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1801 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1861 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1921 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1981 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2041 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2101 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2161 gttagctcct tcggtcctcc gatagttgtc agaagtaagt tggccgcagt gttatcactc
     2221 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2281 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2341 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt gaaagtgctc
     2401 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2461 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2521 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2581 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2641 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2701 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattac gtagaaagcc
     2761 agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg
     2821 gaaaacgcaa gcgcaaagag gatccttatc gctatcgatt cacacaaaaa accaacacac
     2881 agatgtaatg aaaataaaga tattttattg cggccatcgt gatggcaggt tgggcgtcgc
     2941 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     3001 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     3061 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     3121 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     3181 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     3241 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     3301 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     3361 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     3421 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     3481 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     3541 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     3601 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     3661 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     3721 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatccctcc ggagcctccg
     3781 ccacctgatc cgccacctcc gctcccacca ccggtctgct cgttcttcag cacgcgctcc
     3841 acgaagctct tgatgtactt acccatttca tctggagcgt cctcctggct gaagtggagg
     3901 cccttcacct tcacgaactc ggtgttaggg aacttcttag ctccctcgac aatagcgttg
     3961 gaaaagaacc cagggtcgga ctcgatgaac atcttaggca gatcgtcgct ggcccgaagg
     4021 taggcgttgt agttgcggac aatctggacg acgtcgggct tgcctccctt aacgagaggg
     4081 atctcgcgag gccaggagag ggtaggccgt ctaacctcgc ccttctcctt gaatggctcc
     4141 aggtaggcag cgaactcctc aggctccagt ttccgcatga tcttgcttgg gagcatggtc
     4201 tcgacgaaga agttattctc aagcaccatt ttctcgccct cttcgctctt gatcagggcg
     4261 atatcctcct cgatgtcagg ccactcgtcc caggactcga tcacgtccac gacactctca
     4321 gcatggacga tggccttgat cttgtcttgg tgctcgtagg agtagtgaaa ggccagacaa
     4381 gccccccagt cgtggcccac aaagatgatt ttctttggaa ggttcagcag ctcgaaccaa
     4441 gcggtgaggt acttgtagtg atccaggagg cgatatgagc cattcccgct cttgccggac
     4501 ttacccattc cgatcagatc agggatgatg catctagcca cgggctcgat gtgaggcacg
     4561 acgtgcctcc acaggtagct ggaggcagcg ttaccatgca gaaaaatcac ggcgttctcg
     4621 gcgtgcttct cggaatcata gtagttgatg aaggagtcca gcacgttcat ttgcttgcag
     4681 cgagcccacc actgaggccc agtgatcatg cgtttgcgtt gctcggggtc gtacaccttg
     4741 gaagcgtcaa aagccctgta tacgacatca gtactagtgc ctgtgccgca cggtgtaaga
     4801 cgggctgcac ttacaccgca aacccaaacg agttcacggc gatcttgccg cccttcttgg
     4861 ccttaatgag aatctcgcgg atcttgcggg cgtccaactt gccggtcagt cctttaggca
     4921 cctcgtccac gaacacaaca ccaccgcgca gcttcttggc ggttgtaacc tggctggcca
     4981 catagtccac gatctccttc tcggtcatgg ttttaccgtg ttccagcacg acgactgcgg
     5041 cgggcagctc gccggcatcg tcgtcgggca ggccggcgac cccggcgtcg aagatgttgg
     5101 ggtgttgcag caggatgctc tccagttcgg ctggggctac ctggtagccc ttgtatttga
     5161 tcaggctctt cagccggtcc acgatgaaga agtgctcgtc ctcgtcccag taggcgatgt
     5221 cgccgctgtg cagccagccg tccttgtcga tgagagcgtt tgtagcctcg gggttgttaa
     5281 cgtagccgct catgatcatg gggccacgga cgcacagctc gccgcgctgg ttcacaccca
     5341 gtgtcttacc ggtgtccaag tccaccacct tagcctcgaa gaagggcacc accttgccta
     5401 ctgcgccagg cttgtcgtcc ccttcggggg tgatcagaat ggcgctggtt gtttctgtca
     5461 ggccgtagcc ctggcggatg cctggtaggt ggaagcgttt ggccacggcc tcacctacct
     5521 ccttgctgag cggcgccccg ccgctggcga tctcgtgcaa gttgcttagg tcgtacttgt
     5581 cgatgagagt gctcttagcg aagaagctaa atagtgtggg caccagcagg gcagattgaa
     5641 tcttatagtc ttgcaagctg cgcaagaata gctcctcctc gaagcggtac atgagcacga
     5701 cccgaaagcc gcagatcaag tagcccagcg tggtgaacat gccgaagccg tggtgaaatg
     5761 gcaccacgct gaggatagcg gtgtcgggga tgatctggtt gccgaagatg gggtcgcggg
     5821 catgactgaa tcggacacaa gcggtgcggt gcggtagggc tacgcccttg ggcaatccgg
     5881 tactgccact actgttcatg atcagggcga tggttttgtc ccggtcgaag ctctcgggca
     5941 cgaagtcgta ctcgttgaag ccgggtggca aatgggaagt cacgaaggtg tacatgcttt
     6001 ggaagccctg gtagtcggtc ttgctatcca tgatgatgat cttttgtatg atcggtagct
     6061 tcttttgcac gttgaggatc ttttgcagcc ctttcttgct cacgaatacg acggtgggct
     6121 ggctgatgcc catgctgttc agcagctcgc gctcgttgta gatgtcgtta gctggggcca
     6181 cagccacacc gatgaacagg gcacccaaca cgggcatgaa gaactgcaag ctattctcgc
     6241 tgcacaccac gatccgatgg tttgtattca gcccatagcg cttcatagct tctgccagcc
     6301 gaacgctcat ctcgaagtac tcggcgtagg taatgtccac ctcgatatgt gcgtcggtaa
     6361 aggcgatggt gccgggcacc agggcgtagc gcttcatggc tttgtgcagc tgctcgccgg
     6421 cggtcccgtc ttcgagtggg tagaatggcg ctgggccctt cttaatgttt ttggcatctt
     6481 ccatggtggc tt
//
