LOCUS       sequence-342371-        5148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 3C-like proteinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5148)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5148)
  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"
COMMENT     Sequence Label: pGBW-m4133586
FEATURES             Location/Qualifiers
     source          1..5148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1504..1524
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1525..1542
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1595..1716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1606..1625)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1660..1679
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1831..1877
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1907..2043
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2088..2159
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2385..2973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2465..2484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3719..3787,3788..4579)
                     /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     complement(3937..3956)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4592..4728
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4880..4951
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4956..5002)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5020..5068
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgagtgg gttccggaaa
      601 atggcgtttc cctccggtaa ggtcgaagga tgcatggttc aggtgacctg tggcacaact
      661 acgttaaacg gcctatggct ggacgatgtg gtatactgtc ctcgacacgt catctgcaca
      721 tcagaggaca tgctgaatcc aaactatgag gatttgctca taagaaagag caatcataac
      781 tttctggtgc aagctggaaa tgtacagctt agggttattg ggcactctat gcagaactgc
      841 gtgttaaaac tcaaggtgga taccgccaat ccgaaaacac ctaagtacaa attcgtgcgc
      901 atccagccag gccaaacctt ctctgtcctg gcatgttata acggcagccc ctcaggtgtc
      961 taccagtgcg ccatgcgccc taattttact attaagggtt ccttcttgaa cgggtcgtgc
     1021 ggatcggtcg gctttaatat cgactatgac tgtgtgagtt tttgctacat gcatcacatg
     1081 gaacttccga ctggggttca cgctggaacg gatctggagg gtaacttcta tggacccttc
     1141 gtggatagac agaccgccca ggcagccggc actgacacta ccattacggt gaatgttcta
     1201 gcttggctct acgcagctgt aataaacggc gatcgttggt tcttgaacag gtttacaaca
     1261 accctgaacg actttaatct agtcgccatg aaatataatt atgaaccact gacccaagat
     1321 catgtagaca ttctcgggcc actgagcgca cagacaggga tcgctgtgct ggacatgtgt
     1381 gccagtttaa aggagctgct tcagaatggt atgaacggcc gaacaatcct agggtctgcg
     1441 ctcttagagg atgaatttac ccccttcgat gtagtgcggc aatgtagcgg cgttactttt
     1501 caagaaaacc tttactttca gtcgcaccat caccatcacc attaagctag cttgactgac
     1561 tgagatacag cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa
     1621 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1681 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1741 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     1801 tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg
     1861 tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa
     1921 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     1981 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     2041 taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg
     2101 gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa
     2161 tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg
     2221 agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt
     2281 ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc
     2341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2581 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2821 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3001 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3061 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3181 atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg
     3241 tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta
     3301 tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg
     3361 tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc
     3421 ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc ctggtgtacc
     3481 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     3541 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     3601 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     3661 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     3721 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3781 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     3841 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3901 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3961 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4021 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4081 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     4141 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4201 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4261 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     4321 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4381 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4441 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4501 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4561 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     4621 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     4681 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     4741 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     4801 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     4861 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     4921 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     4981 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     5041 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     5101 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
