LOCUS       sequence-420347-        7524 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 7524)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7524)
  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-m4133568
FEATURES             Location/Qualifiers
     source          1..7524
                     /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..1566
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1635..1756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1646..1665)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1700..1719
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1871..1917
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1947..2083
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2128..2199
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2425..3013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2505..2524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3513..4722
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4559..4578
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4726..5754
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5781..5829
                     /note="synthetic polyadenylation signal"
     CDS             join(6095..6163,6164..6955)
                     /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(6313..6332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6968..7104
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7256..7327
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7332..7378)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7396..7444
                     /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 gtcggggaaa cctataccca atccacttct cggcttagat
     1561 tctacctaag gatccgcagg cctctgctag cttgactgac tgagatacag cgtaccttca
     1621 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1681 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1741 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1801 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
     1861 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
     1921 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
     1981 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     2041 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
     2101 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
     2161 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
     2221 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     2281 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     2341 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2401 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2461 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2521 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2581 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2641 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2701 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2761 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2821 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2881 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2941 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3001 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3061 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3121 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3181 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3241 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     3301 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     3361 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     3421 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     3481 gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc
     3541 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     3601 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     3661 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     3721 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     3781 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     3841 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     3901 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     3961 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     4021 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     4081 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     4141 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     4201 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     4261 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     4321 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     4381 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     4441 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     4501 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     4561 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     4621 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     4681 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg taaaaagcct
     4741 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
     4801 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
     4861 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
     4921 cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg ggagttcagc
     4981 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
     5041 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc catcgctgcc
     5101 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
     5161 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     5221 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     5281 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     5341 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     5401 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     5461 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg gctccgggcg
     5521 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     5581 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     5641 gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg tgtagaagta
     5701 ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga ataagctagt
     5761 atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
     5821 ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     5881 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     5941 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     6001 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     6061 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     6121 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6181 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     6241 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     6301 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6361 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     6421 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6481 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     6541 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6601 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     6661 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     6721 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6781 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6841 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6901 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     6961 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     7021 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     7081 tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc tggctctatc
     7141 gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat
     7201 cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct
     7261 tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct
     7321 acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc
     7381 aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     7441 tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt
     7501 tcatagccca tatatggagt tccg
//
