LOCUS       sequence-295151-        7511 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 7511)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7511)
  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-m4133414
FEATURES             Location/Qualifiers
     source          1..7511
                     /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    1051..1172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1062..1081)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1116..1135
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1287..1333
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1363..1499
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1544..1615
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1841..2429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1921..1940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2929..4138
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3975..3994
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4142..5170
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5197..5245
                     /note="synthetic polyadenylation signal"
     CDS             join(5511..5579,5580..6371)
                     /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(5729..5748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6384..6520
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6665..6736
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6741..6787)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6805..6853
                     /note="synthetic polyadenylation signal"
     enhancer        6934..7237
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7239..7441
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7392..7412
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7510..7511,1..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 cctaaggatc cgcaggcctc tgctagcttg actgactgag
     1021 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca
     1081 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1141 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1201 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     1261 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt
     1321 gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa
     1381 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     1441 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     1501 gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct
     1561 tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc
     1621 attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat
     1681 cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt
     1741 gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc
     1801 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1861 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1921 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1981 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2041 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2101 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2161 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2221 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2281 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2341 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2641 gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg
     2701 aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc
     2761 aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt
     2821 actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca
     2881 atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc
     2941 gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg
     3001 aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc
     3061 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg
     3121 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct
     3181 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga
     3241 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt
     3301 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc
     3361 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt
     3421 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg
     3481 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca
     3541 aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga
     3601 aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt
     3661 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
     3721 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
     3781 tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct
     3841 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
     3901 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
     3961 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
     4021 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
     4081 gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa
     4141 catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     4201 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     4261 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     4321 agatcgttat gtttatcggc actttgcatc cgccgcgctc ccgattccgg aagtgcttga
     4381 cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     4441 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggcaat
     4501 ggatgccatc gctgccgccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     4561 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     4621 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     4681 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     4741 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     4801 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     4861 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     4921 gccccggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     4981 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     5041 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgccgccg tctggaccga
     5101 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     5161 aaaggaataa gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga
     5221 tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga
     5281 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     5341 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     5401 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     5461 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     5521 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5581 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     5641 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     5701 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5761 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     5821 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5881 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     5941 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6001 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6061 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6121 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6181 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6241 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6301 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6361 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     6421 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     6481 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcgtttaa actcgactct
     6541 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     6601 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     6661 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     6721 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     6781 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     6841 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     6901 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     6961 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     7021 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     7081 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     7141 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     7201 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
     7261 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     7321 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     7381 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     7441 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
     7501 ccagcggccg c
//
