LOCUS       sequence-295678-        8024 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 8024)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8024)
  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-m4134203
FEATURES             Location/Qualifiers
     source          1..8024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             920..940
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             974..1051
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1079..1099
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1564..1685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1575..1594)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1629..1648
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1800..1846
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1876..2012
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2057..2128
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2354..2942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2434..2453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3442..4651
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4488..4507
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4655..5683
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5710..5758
                     /note="synthetic polyadenylation signal"
     CDS             join(6024..6092,6093..6884)
                     /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(6242..6261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6897..7033
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7178..7249
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7254..7300)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7318..7366
                     /note="synthetic polyadenylation signal"
     enhancer        7447..7750
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7752..7954
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7905..7925
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8023..8024,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggt tttagaaaaa tggcattccc atctggtaaa gttgagggtt gtatggtaca
       61 agtaacttgt ggtacaacta cacttaacgg tctttggctt gatgacgtag tttactgtcc
      121 aagacatgtg atctgcacct ctgaagacat gcttaaccct aattatgaag atttactcat
      181 tcgtaagtct aatcataatt tcttggtaca ggctggtaat gttcaactca gggttattgg
      241 acattctatg caaaattgtg tacttaagct taaggttgat acagccaatc ctaagacacc
      301 taagtataag tttgttcgca ttcaaccagg acagactttt tcagtgttag cttgttacaa
      361 tggttcacca tctggtgttt accaatgtgc tatgaggccc aatttcacta ttaagggttc
      421 attccttaat ggttcatgtg gtagtgttgg ttttaacata gattatgact gtgtctcttt
      481 ttgttacatg caccatatgg aattaccaac tggagttcat gctggcacag acttagaagg
      541 taacttttat ggaccttttg ttgacaggca aacagcacaa gcagctggta cggacacaac
      601 tattacagtt aatgttttag cttggttgta cgctgctgtt ataaatggag acaggtggtt
      661 tctcaatcga tttaccacaa ctcttaatga ctttaacctt gtggctatga agtacaatta
      721 tgaacctcta acacaagacc atgttgacat actaggacct ctttctgctc aaactggaat
      781 tgccgtttta gatatgtgtg cttcattaaa agaattactg caaaatggta tgaatggacg
      841 taccatattg ggtagtgctt tattagaaga tgaatttaca ccttttgatg ttgttagaca
      901 atgctcaggt gttactttcg aaaaccttta ctttcagtcg ggacggcgaa tcccggggct
      961 gataaatccc tggaaaagga gatggaagaa gaactttatt gcagtgtctg cggctaaccg
     1021 gttcaaaaag atcagttcaa gcggcgccct agattacgac attcctacga ccgcctccga
     1081 gaatctctat ttccagggtg aatttggctt ggcacaacat gacgaggccg tcgataacaa
     1141 gtttaataaa gaacagcaga acgctttcta tgagatcctt cacttaccaa atctgaacga
     1201 agagcaacgc aacgctttca ttcagagcct gaaagacgat ccctcgcagt ctgcaaatct
     1261 cctggccgaa gccaagaagc tgaatgacgc acaggctcca aaagttgata acaagtttaa
     1321 caaggagcag cagaatgctt tctacgagat attgcacctt cctaatctga atgaggaaca
     1381 aagaaatgca tttatccagt ccctaaaaga cgatccatca cagagtgcca acctgcttgc
     1441 ggaagccaaa aagctcaatg atgcccaggc cccgaaagta gacgcgaacc atcaataagg
     1501 atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca
     1561 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1621 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1681 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1741 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
     1801 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
     1861 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
     1921 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
     1981 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
     2041 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
     2101 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
     2161 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
     2221 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     2281 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2341 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2401 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2461 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2521 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2581 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2641 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2701 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2761 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2821 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2881 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     3181 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     3241 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     3301 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     3361 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     3421 tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     3481 ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc
     3541 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     3601 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     3661 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     3721 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     3781 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     3841 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     3901 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     3961 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     4021 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     4081 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     4141 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     4201 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     4261 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     4321 ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     4381 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     4441 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     4501 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     4561 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
     4621 attctggccg tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc
     4681 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     4741 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     4801 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     4861 atccgccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac
     4921 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     4981 gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag
     5041 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     5101 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     5161 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     5221 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     5281 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     5341 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     5401 cttcgagcgg aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg
     5461 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     5521 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     5581 aatcgcccgc agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     5641 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta
     5701 gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg
     5761 taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     5821 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     5881 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     5941 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     6001 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     6061 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6121 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     6181 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     6241 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6301 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     6361 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6421 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     6481 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6541 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     6601 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     6661 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6721 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6781 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6841 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     6901 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     6961 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     7021 aatttatcct taagatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     7081 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     7141 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     7201 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     7261 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     7321 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     7381 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     7441 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     7501 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     7561 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     7621 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     7681 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     7741 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     7801 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     7861 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     7921 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc
     7981 tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
