LOCUS       sequence-295864-        5166 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 5166)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5166)
  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-m4133410
FEATURES             Location/Qualifiers
     source          1..5166
                     /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             941..982
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1035..1156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1046..1065)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1100..1119
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1271..1317
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1347..1483
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1528..1599
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1825..2413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1905..1924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3159..3227,3228..4019)
                     /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(3377..3396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4032..4168
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4320..4391
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4396..4442)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4460..4508
                     /note="synthetic polyadenylation signal"
     enhancer        4589..4892
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4894..5096
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5047..5067
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5165..5166,1..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 gggaaaccta tacccaatcc
      961 acttctcggc ttagattcta cctaagctag cttgactgac tgagatacag cgtaccttca
     1021 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1081 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1141 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1201 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
     1261 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
     1321 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
     1381 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     1441 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
     1501 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
     1561 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
     1621 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     1681 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     1741 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1801 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1861 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1921 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1981 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2041 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2101 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2161 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2221 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2281 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2341 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2401 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2461 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2521 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2581 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2641 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     2701 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     2761 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     2821 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     2881 gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     2941 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     3001 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     3061 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     3121 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     3181 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3241 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     3301 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     3361 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3421 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     3481 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3541 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     3601 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3661 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     3721 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     3781 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3841 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3901 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3961 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4021 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     4081 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     4141 gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc
     4201 tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg
     4261 gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac
     4321 atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc
     4381 agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
     4441 tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg
     4501 attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat
     4561 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     4621 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     4681 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4741 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4801 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     4861 acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg
     4921 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     4981 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     5041 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
     5101 tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc
     5161 ggccgc
//
