LOCUS       sequence-310881-        5193 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 5193)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5193)
  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-m4133721
FEATURES             Location/Qualifiers
     source          1..5193
                     /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"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1498..1518
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1522..1587
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1640..1761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1651..1670)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1705..1724
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1876..1922
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1952..2088
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2133..2204
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2430..3018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2510..2529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3764..3832,3833..4624)
                     /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(3982..4001)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4637..4773
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4925..4996
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5001..5047)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5065..5113
                     /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 ccatgggttt tagaaaaatg
      601 gcattcccat ctggtaaagt tgagggttgt atggtacaag taacttgtgg tacaactaca
      661 cttaacggtc tttggcttga tgacgtagtt tactgtccaa gacatgtgat ctgcacctct
      721 gaagacatgc ttaaccctaa ttatgaagat ttactcattc gtaagtctaa tcataatttc
      781 ttggtacagg ctggtaatgt tcaactcagg gttattggac attctatgca aaattgtgta
      841 cttaagctta aggttgatac agccaatcct aagacaccta agtataagtt tgttcgcatt
      901 caaccaggac agactttttc agtgttagct tgttacaatg gttcaccatc tggtgtttac
      961 caatgtgcta tgaggcccaa tttcactatt aagggttcat tccttaatgg ttcatgtggt
     1021 agtgttggtt ttaacataga ttatgactgt gtctcttttt gttacatgca ccatatggaa
     1081 ttaccaactg gagttcatgc tggcacagac ttagaaggta acttttatgg accttttgtt
     1141 gacaggcaaa cagcacaagc agctggtacg gacacaacta ttacagttaa tgttttagct
     1201 tggttgtacg ctgctgttat aaatggagac aggtggtttc tcaatcgatt taccacaact
     1261 cttaatgact ttaaccttgt ggctatgaag tacaattatg aacctctaac acaagaccat
     1321 gttgacatac taggacctct ttctgctcaa actggaattg ccgttttaga tatgtgtgct
     1381 tcattaaaag aattactgca aaatggtatg aatggacgta ccatattggg tagtgcttta
     1441 ttagaagatg aatttacacc ttttgatgtt gttagacaat gctcaggtgt tactttcgaa
     1501 aacctttact ttcagtcgat ggattataag gaccacgatg gagactacaa agatcatgac
     1561 attgattaca aggacgacga tgacaaataa gctagcttga ctgactgaga tacagcgtac
     1621 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1681 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1741 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1801 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     1861 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1921 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     1981 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     2041 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     2101 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     2161 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     2221 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     2281 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     2341 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     2401 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2461 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2521 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2581 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2641 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2701 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2761 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2821 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2881 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2941 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3001 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3061 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3121 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3181 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3241 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     3301 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     3361 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     3421 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     3481 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc
     3541 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     3601 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     3661 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     3721 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     3781 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3841 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     3901 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     3961 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4021 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     4081 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4141 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     4201 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4261 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4321 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     4381 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4441 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4501 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4561 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4621 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     4681 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     4741 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     4801 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     4861 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     4921 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     4981 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     5041 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     5101 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     5161 gtcattagtt catagcccat atatggagtt ccg
//
