LOCUS       sequence-420912-        4241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For assessing leaky scanning rates in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4241)
  TITLE     The SARS-CoV-2 protein ORF3c is a mitochondrial modulator of innate 
            immunity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4241)
  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: 
            "Unpublished"
COMMENT     Sequence Label: p_L_3c_SG_RLuc
FEATURES             Location/Qualifiers
     source          1..4241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(68..859,860..928))
                     /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    1033..1081
                     /note="synthetic polyadenylation signal"
     misc_feature    1095..1186
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1135..1154
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1292..1649
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1292..1483
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1315..1335)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1500..1635
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        1656..1674
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1939..2004
                     /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             2005..2937
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="ASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2981..3102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2992..3011)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3046..3065
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3350..3367)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3521..4109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3601..3620)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatctaaa gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact
       61 gcagtggtta ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt
      121 cgtccatagt ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat
      181 caggccccag cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag
      241 caatgaacca gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct
      301 ccatccagtc tatgagctgc tgtcttgatg ctagagtaag aagttcgcca gtgagtagtt
      361 tccgaagagt tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg
      421 cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa
      481 gaaatgcagt cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt
      541 tgtcgctcat ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat
      601 gcttttccgt gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac
      661 caagctgctc ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga
      721 aagtgctcat catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat
      781 tgagatccag ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt
      841 tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga
      901 gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt
      961 atcagggtta ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat
     1021 tggacaggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     1081 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     1141 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctggcc taactggccg
     1201 gtacctgagc tcgctagctt cgaggatatc agatcagcgc agcaccatgg cctgaaataa
     1261 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
     1321 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1381 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     1441 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     1501 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     1561 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     1621 gcttttttgg aggcctaggc ttttgcaaaa agctctaata cgactcacta taggattaaa
     1681 ggtttatacc ttcccaggta acaaaccaac caactttcga tctcttgtag atctgttctc
     1741 taaacgaact tatggatttg tttatgagaa tcttcacaat tggaactgta actttgaagc
     1801 aaggtgaaat caaggatgct actccttcag attttgttcg cgctactgca acgataccga
     1861 tacaagcctc actccctttc ggatggctta ttgttggcgt tgcacttctt gctgtttttc
     1921 agagcgcttc caaaatcagt gaaacagact ttgaattttg accttcttaa gctggcggga
     1981 gacgtcgagt ccaaccccgg gcccgcttcc aaggtgtacg accccgagca acgcaaacgc
     2041 atgatcactg ggcctcagtg gtgggctcgc tgcaagcaaa tgaacgtgct ggactccttc
     2101 atcaactact atgattccga gaagcacgcc gagaacgccg tgatttttct gcatggtaac
     2161 gctgcctcca gctacctgtg gaggcacgtc gtgcctcaca tcgagcccgt ggctagatgc
     2221 atcatccctg atctgatcgg aatgggcaaa tccggcaaga gcgggaatgg ctcatatcgc
     2281 ctcctggatc actacaagta cctcaccgct tggttcgagc tgctgaacct tccaaagaaa
     2341 atcatctttg tgggccacga ctggggggct tgtctggcct ttcactactc ctacgagcac
     2401 caagacaaga tcaaggccat cgtccatgct gagagtgtcg tggacgtgat cgagtcctgg
     2461 gacgagtggc ctgacatcga ggaggatatc gccctgatca agagcgaaga gggcgagaaa
     2521 atggtgcttg agaataactt cttcgtcgag accatgctcc caagcaagat catgcggaaa
     2581 ctggagcctg aggagttcgc tgcctacctg gagccattca aggagaaggg cgaggttaga
     2641 cggcctaccc tctcctggcc tcgcgagatc cctctcgtta agggaggcaa gcccgacgtc
     2701 gtccagattg tccgcaacta caacgcctac cttcgggcca gcgacgatct gcctaagatg
     2761 ttcatcgagt ccgaccctgg gttcttttcc aacgctattg tcgagggagc taagaagttc
     2821 cctaacaccg agttcgtgaa ggtgaagggc ctccacttca gccaggagga cgctccagat
     2881 gaaatgggaa aatacatcaa gagcttcgtg gagcgcgtgc tgaagaacga gcagtaatga
     2941 ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     3001 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3061 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3121 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     3181 tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc
     3241 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     3301 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     3361 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3421 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3961 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4021 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4081 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4141 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4201 tcttcaccta gatcctttta aattaaaaat gaagttttaa a
//
