LOCUS       sequence-226674-        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"
     promoter        106..463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        106..297
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(129..149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      314..449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     470..489
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        470..488
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      518..527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             524..1456
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             1508..1573
                     /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             1630..1695
                     /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             1714..3363
                     /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    3407..3528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3418..3437)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3472..3491
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3776..3793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3947..4535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4027..4046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4735..5526,5527..5595))
                     /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    5700..5748
                     /note="synthetic polyadenylation signal"
     misc_feature    5762..5853
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5802..5821
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcttcgagga tatcagatca gcgcagcacc
       61 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
      121 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
      181 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
      241 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
      301 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
      361 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
      421 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctct aatacgactc
      481 actataggga gcttggcaat ccggtactgt tggtaaagcc accatggctt ccaaggtgta
      541 cgaccccgag caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca
      601 aatgaacgtg ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc
      661 cgtgattttt ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca
      721 catcgagccc gtggctagat gcatcatccc tgatctgatc ggaatgggca aatccggcaa
      781 gagcgggaat ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga
      841 gctgctgaac cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc
      901 ctttcactac tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt
      961 cgtggacgtg atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat
     1021 caagagcgaa gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct
     1081 cccaagcaag atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt
     1141 caaggagaag ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt
     1201 taagggaggc aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc
     1261 cagcgacgat ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat
     1321 tgtcgaggga gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt
     1381 cagccaggag gacgctccag atgaaatggg aaaatacatc aagagcttcg tggagcgcgt
     1441 gctgaagaac gagcagtatt tctacacctc gagagaagcc agacacaaac agaaaattgt
     1501 ggcaccggtg aaacagactt tgaattttga ccttcttaag ctggcgggag acgtcgagtc
     1561 caaccccggg ccctactcga gcagaagctt agatctgagg cacggcataa gcaaaagatc
     1621 gtagccccag taaagcaaac actcaacttc gatctactca aactcgcagg tgatgtggaa
     1681 tctaatccag gacctttcgg atctgccacc attgaagatg ccaaaaacat taagaagggc
     1741 ccagcgccat tctacccact cgaagacggg accgccggcg agcagctgca caaagccatg
     1801 aagcgctacg ccctggtgcc cggcaccatc gcctttaccg acgcacatat cgaggtggac
     1861 attacctacg ccgagtactt cgagatgagc gttcggctgg cagaagctat gaagcgctat
     1921 gggctgaata caaaccatcg gatcgtggtg tgcagcgaga atagcttgca gttcttcatg
     1981 cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc cagctaacga catctacaac
     2041 gagcgcgagc tgctgaacag catgggcatc agccagccca ccgtcgtatt cgtgagcaag
     2101 aaagggctgc aaaagatcct caacgtgcaa aagaagctac cgatcataca aaagatcatc
     2161 atcatggata gcaagaccga ctaccagggc ttccaaagca tgtacacctt cgtgacttcc
     2221 catttgccac ccggcttcaa cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa
     2281 accatcgccc tgatcatgaa cagtagtggc agtaccggat tgcccaaggg cgtagcccta
     2341 ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg accccatctt cggcaaccag
     2401 atcatccccg acaccgctat cctcagcgtg gtgccatttc accacggctt cggcatgttc
     2461 accacgctgg gctacttgat ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag
     2521 gagctattct tgcgcagctt gcaagactat aagattcaat ctgccctgct ggtgcccaca
     2581 ctatttagct tcttcgctaa gagcactctc atcgacaagt acgacctaag caacttgcac
     2641 gagatcgcca gcggcggggc gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc
     2701 ttccacctac caggcatccg ccagggctac ggcctgacag aaacaaccag cgccattctg
     2761 atcacccccg aaggggacga caagcctggc gcagtaggca aggtggtgcc cttcttcgag
     2821 gctaaggtgg tggacttgga caccggtaag acactgggtg tgaaccagcg cggcgagctg
     2881 tgcgtccgtg gccccatgat catgagcggc tacgttaaca accccgaggc tacaaacgct
     2941 ctcatcgaca aggacggctg gctgcacagc ggcgacatcg cctactggga cgaggacgag
     3001 cacttcttca tcgtggaccg gctgaagagc ctgatcaaat acaagggcta ccaggtagcc
     3061 ccagccgaac tggagagcat cctgctgcaa caccccaaca tcttcgacgc cggggtcgcc
     3121 ggcctgcccg acgacgatgc cggcgagctg cccgccgcag tcgtcgtgct ggaacacggt
     3181 aaaaccatga ccgagaagga gatcgtggac tatgtggcca gccaggttac aaccgccaag
     3241 aagctgcgcg gtggtgttgt gttcgtggac gaggtgccta aaggactgac cggcaagttg
     3301 gacgcccgca agatccgcga gattctcatt aaggccaaga agggcggcaa gatcgccgtg
     3361 taataattct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     3421 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3481 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3541 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     3601 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     3661 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     3721 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     3781 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3841 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3901 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3961 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4021 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4081 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4141 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4201 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4261 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4321 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4381 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4441 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4501 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4561 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4621 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4681 tatatgagta aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca
     4741 gtgcttgatc agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc
     4801 ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc
     4861 agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc
     4921 agcagggagg gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat
     4981 gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt
     5041 ggccattgct actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc
     5101 tggttcccag cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag
     5161 ctccttaggg cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt
     5221 aatggcagca ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac
     5281 cggcgagtac tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg
     5341 cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat
     5401 cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc
     5461 gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc
     5521 ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa
     5581 atgttggatg ctcatactcg tcctttttca atattattga agcatttatc agggttacta
     5641 gtacgtctct caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca
     5701 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
     5761 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
     5821 ccagtgcaag tgcaggtgcc agaacatttc tct
//
