LOCUS       sequence-233131-        5853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dual luciferase with FMDV StopGo sequences flanking MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  AUTHORS   Loughran G, Fedorova AD, Khan YA, Atkins JF, Baranov PV
  TITLE     Lack of evidence for ribosomal frameshifting in ATP7B mRNA decoding.
  JOURNAL   Mol Cell. 2022 Oct 6;82(19):3745-3749.e2. doi: 
            10.1016/j.molcel.2022.08.024. Epub 2022 Sep 16.
  PUBMED    36115342
REFERENCE   2  (bases 1 to 5853)
  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"; doi: 
            "10.1016/j.molcel.2022.08.024"; journalName: "Mol Cell"; date: 
            "2022-10-6- 6"; volume: "82"; issue: "19"; pages: "3745-3749.e2"
COMMENT     Sequence Label: pSGDlucV3.0
FEATURES             Location/Qualifiers
     source          1..5853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..93
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        74..92
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      122..131
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             128..1060
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             1112..1177
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             1234..1299
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             1318..2967
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    3011..3132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3022..3041)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3076..3095
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3380..3397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3551..4139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3631..3650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4339..5130,5131..5199))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5304..5352
                     /note="synthetic polyadenylation signal"
     misc_feature    5366..5457
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5406..5425
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        join(5563..5853,1..67)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5563..5754
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5586..5606)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5771..5853,1..53)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
       61 ttgcaaaaag ctctaatacg actcactata gggagcttgg caatccggta ctgttggtaa
      121 agccaccatg gcttccaagg tgtacgaccc cgagcaacgc aaacgcatga tcactgggcc
      181 tcagtggtgg gctcgctgca agcaaatgaa cgtgctggac tccttcatca actactatga
      241 ttccgagaag cacgccgaga acgccgtgat ttttctgcat ggtaacgctg cctccagcta
      301 cctgtggagg cacgtcgtgc ctcacatcga gcccgtggct agatgcatca tccctgatct
      361 gatcggaatg ggcaaatccg gcaagagcgg gaatggctca tatcgcctcc tggatcacta
      421 caagtacctc accgcttggt tcgagctgct gaaccttcca aagaaaatca tctttgtggg
      481 ccacgactgg ggggcttgtc tggcctttca ctactcctac gagcaccaag acaagatcaa
      541 ggccatcgtc catgctgaga gtgtcgtgga cgtgatcgag tcctgggacg agtggcctga
      601 catcgaggag gatatcgccc tgatcaagag cgaagagggc gagaaaatgg tgcttgagaa
      661 taacttcttc gtcgagacca tgctcccaag caagatcatg cggaaactgg agcctgagga
      721 gttcgctgcc tacctggagc cattcaagga gaagggcgag gttagacggc ctaccctctc
      781 ctggcctcgc gagatccctc tcgttaaggg aggcaagccc gacgtcgtcc agattgtccg
      841 caactacaac gcctaccttc gggccagcga cgatctgcct aagatgttca tcgagtccga
      901 ccctgggttc ttttccaacg ctattgtcga gggagctaag aagttcccta acaccgagtt
      961 cgtgaaggtg aagggcctcc acttcagcca ggaggacgct ccagatgaaa tgggaaaata
     1021 catcaagagc ttcgtggagc gcgtgctgaa gaacgagcag tatttctaca cctcgagaga
     1081 agccagacac aaacagaaaa ttgtggcacc ggtgaaacag actttgaatt ttgaccttct
     1141 taagctggcg ggagacgtcg agtccaaccc cgggccctac tcgagcagaa gcttagatct
     1201 gaggcacggc ataagcaaaa gatcgtagcc ccagtaaagc aaacactcaa cttcgatcta
     1261 ctcaaactcg caggtgatgt ggaatctaat ccaggacctt tcggatctgc caccattgaa
     1321 gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga cgggaccgcc
     1381 ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac catcgccttt
     1441 accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat gagcgttcgg
     1501 ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc
     1561 gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg
     1621 gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg catcagccag
     1681 cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag
     1741 ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca gggcttccaa
     1801 agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta cgacttcgtg
     1861 cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag tggcagtacc
     1921 ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc
     1981 cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag cgtggtgcca
     2041 tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg ctttcgggtc
     2101 gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga ctataagatt
     2161 caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac tctcatcgac
     2221 aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct cagcaaggag
     2281 gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg ctacggcctg
     2341 acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc tggcgcagta
     2401 ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg taagacactg
     2461 ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt
     2521 aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca cagcggcgac
     2581 atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa gagcctgatc
     2641 aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct gcaacacccc
     2701 aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga gctgcccgcc
     2761 gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt ggactatgtg
     2821 gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg
     2881 cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct cattaaggcc
     2941 aagaagggcg gcaagatcgc cgtgtaataa ttctagagtc ggggcggccg gccgcttcga
     3001 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3061 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3121 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3181 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg
     3241 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     3301 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     3361 ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3421 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3481 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3541 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3601 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3661 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3721 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3781 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3841 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3901 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3961 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4021 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4081 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4141 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4201 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4261 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc ggccgcaaat
     4321 gctaaaccac tgcagtggtt accagtgctt gatcagtgag gcaccgatct cagcgatctg
     4381 cctatttcgt tcgtccatag tggcctgact ccccgtcgtg tagatcacta cgattcgtga
     4441 gggcttacca tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt caccggcccc
     4501 cgatttgtca gcaatgaacc agccagcagg gagggccgag cgaagaagtg gtcctgctac
     4561 tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc
     4621 agtgagtagt ttccgaagag ttgtggccat tgctactggc atcgtggtat cacgctcgtc
     4681 gttcggtatg gcttcgttca actctggttc ccagcggtca agccgggtca catgatcacc
     4741 catattatga agaaatgcag tcagctcctt agggcctccg atcgttgtca gaagtaagtt
     4801 ggccgcggtg ttgtcgctca tggtaatggc agcactacac aattctctta ccgtcatgcc
     4861 atccgtaaga tgcttttccg tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg
     4921 tatacggcga ccaagctgct cttgcccggc gtctatacgg gacaacaccg cgccacatag
     4981 cagtactttg aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag actcaaggat
     5041 cttgccgcta ttgagatcca gttcgatata gcccactctt gcacccagtt gatcttcagc
     5101 atcttttact ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa
     5161 gaagggaatg agtgcgacac gaaaatgttg gatgctcata ctcgtccttt ttcaatatta
     5221 ttgaagcatt tatcagggtt actagtacgt ctctcaagga taagtaagta atattaaggt
     5281 acgggaggta ttggacaggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     5341 gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa
     5401 acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctggc
     5461 ctaactggcc ggtacctgag ctcgctagct tcgaggatat cagatcagcg cagcaccatg
     5521 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     5581 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     5641 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     5701 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     5761 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5821 actaattttt tttatttatg cagaggccga ggc
//
