LOCUS       sequence-392885-        5799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal Myc-tagged human codon optimyzed SARS-CoV-2 3CL
            C145A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5799)
  AUTHORS   Arakawa M, Yoshida A, Okamura S, Ebina H, Morita E
  TITLE     A highly sensitive NanoLuc-based protease biosensor for detecting 
            apoptosis and SARS-CoV-2 infection.
  JOURNAL   Sci Rep. 2023 Jan 31;13(1):1753. doi: 10.1038/s41598-023-28984-4.
  PUBMED    36720982
REFERENCE   2  (bases 1 to 5799)
  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: "Sci Rep."; 
            date: "2023-01-31"; volume: "13(1):1753. doi"; pages: " 
            10.1038/s41598-023-28984-4"
COMMENT     Sequence Label: pCAG.Myc-SARS2_3CL(C145A)
FEATURES             Location/Qualifiers
     source          1..5799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1763..1772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1772..1801
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2821..2840)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2886..2941
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2940..2959)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3302..3318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3302..3318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3315..3337)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3326..3342
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3350..3356,3357..3374,3375..3380))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3395..3416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3475..3670
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3521..3656
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3676..3810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3713..3732)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3767..3786
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3878..3895)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4049..4637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4129..4148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4808..5599,5600..5668))
                     /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     5431..5450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5669..5773)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
      541 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgat
     1741 cgacggtatc gataagcttg gcgccaccat ggagcagaag ctgatcagcg aggaggacct
     1801 gggtaccatg tccggattcc gcaagatggc cttcccttct ggcaaggtgg agggctgcat
     1861 ggtgcaggtg acctgcggaa ccaccaccct gaacggtctg tggctggacg acgtggtgta
     1921 ctgccccagg cacgtgatct gcacctctga ggacatgctg aaccctaact acgaagacct
     1981 gctgatcaga aagagcaacc acaacttcct ggtgcaggcc ggtaacgtgc agctgcgtgt
     2041 gatcggccac tctatgcaga actgcgtgct gaagctgaag gtggacaccg ctaaccccaa
     2101 gacccctaag tacaagttcg tgagaatcca gcctggtcag accttctccg tgctggcctg
     2161 ctacaacgga agcccatccg gcgtgtacca gtgcgctatg cgtcccaact tcaccatcaa
     2221 gggcagcttc ctgaacggct ctgccggaag cgtgggtttc aacatcgact acgactgcgt
     2281 gtccttctgc tacatgcacc acatggagct gccaactgga gtgcacgctg gaactgacct
     2341 ggagggtaac ttctacggcc ccttcgtgga cagacagact gctcaggctg ctggcaccga
     2401 caccaccatc accgtgaacg tgctggcttg gctgtacgcc gctgtgatca acggagaccg
     2461 ctggttcctg aacaggttca ccaccaccct gaacgacttc aacctggtgg ctatgaagta
     2521 caactacgag cctctgaccc aggaccacgt ggatatcctg ggtccactga gcgcccagac
     2581 tggaatcgct gtgctggaca tgtgcgcctc cctgaaggaa ctgctgcaga acggcatgaa
     2641 cggacgcacc atcctgggat ccgctctgct ggaggacgaa ttcaccccct tcgacgtggt
     2701 gaggcagtgc tctggtgtga ccttccagta actcgaggaa ttcactcctc aggtgcaggc
     2761 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     2821 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     2881 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     2941 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     3001 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     3061 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     3121 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     3181 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     3241 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
     3301 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3361 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3421 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct
     3481 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3541 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3601 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3661 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3721 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3781 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg
     3841 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3901 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3961 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4021 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4081 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4141 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4201 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4261 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4321 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4381 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4441 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4501 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4561 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4621 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4681 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4741 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4801 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4861 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4921 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4981 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5041 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5101 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5161 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5221 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5281 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5341 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5401 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5461 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5521 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5581 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5641 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5701 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5761 taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
