LOCUS       sequence-295758-        7865 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 7865)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7865)
  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-m4133650
FEATURES             Location/Qualifiers
     source          1..7865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             926..946
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1295..1312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1405..1526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1416..1435)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1470..1489
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1641..1687
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1717..1853
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1898..1969
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2195..2783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2275..2294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3283..4492
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4329..4348
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4496..5524
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5551..5599
                     /note="synthetic polyadenylation signal"
     CDS             join(5865..5933,5934..6725)
                     /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(6083..6102)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6738..6874
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7019..7090
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7095..7141)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7159..7207
                     /note="synthetic polyadenylation signal"
     enhancer        7288..7591
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7593..7795
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7746..7766
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gggttccgga aaatggcgtt tccctccggt aaggtcgaag gatgcatggt
       61 tcaggtgacc tgtggcacaa ctacgttaaa cggcctatgg ctggacgatg tggtatactg
      121 tcctcgacac gtcatctgca catcagagga catgctgaat ccaaactatg aggatttgct
      181 cataagaaag agcaatcata actttctggt gcaagctgga aatgtacagc ttagggttat
      241 tgggcactct atgcagaact gcgtgttaaa actcaaggtg gataccgcca atccgaaaac
      301 acctaagtac aaattcgtgc gcatccagcc aggccaaacc ttctctgtcc tggcatgtta
      361 taacggcagc ccctcaggtg tctaccagtg cgccatgcgc cctaatttta ctattaaggg
      421 ttccttcttg aacgggtcgt gcggatcggt cggctttaat atcgactatg actgtgtgag
      481 tttttgctac atgcatcaca tggaacttcc gactggggtt cacgctggaa cggatctgga
      541 gggtaacttc tatggaccct tcgtggatag acagaccgcc caggcagccg gcactgacac
      601 taccattacg gtgaatgttc tagcttggct ctacgcagct gtaataaacg gcgatcgttg
      661 gttcttgaac aggtttacaa caaccctgaa cgactttaat ctagtcgcca tgaaatataa
      721 ttatgaacca ctgacccaag atcatgtaga cattctcggg ccactgagcg cacagacagg
      781 gatcgctgtg ctggacatgt gtgccagttt aaaggagctg cttcagaatg gtatgaacgg
      841 ccgaacaatc ctagggtctg cgctcttaga ggatgaattt acccccttcg atgtagtgcg
      901 gcaatgtagc ggcgttactt ttcaagaaaa cctttacttt cagtcgatga gtgacaaaat
      961 cattcatcta accgatgact cctttgatac agacgtgttg aaggccgatg gtgcgatact
     1021 ggtagatttc tgggctgaat ggtgtggccc gtgcaagatg atcgcaccca ttctggacga
     1081 gatcgccgac gagtatcaag gaaaattaac tgtcgctaag cttaatatag atcagaaccc
     1141 agggacggca cctaaatacg gcatcagggg aatacctacc ctcctgctct ttaagaatgg
     1201 ggaagttgcc gcgacaaaag tgggagccct gtctaagggc cagctaaaag agttcttgga
     1261 cgctaacctg gcaggttcag gctcggggca catgcaccat caccatcacc atagcagcgg
     1321 gctcgtgcca cgcggctaag gatccgcagg cctctgctag cttgactgac tgagatacag
     1381 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     1441 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1501 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1561 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     1621 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1681 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     1741 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     1801 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     1861 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     1921 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     1981 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     2041 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     2101 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     2161 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2221 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2281 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2341 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2401 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2461 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2521 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2581 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2641 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2701 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2761 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2821 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2881 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2941 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3001 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     3061 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     3121 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     3181 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     3241 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     3301 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     3361 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     3421 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     3481 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     3541 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     3601 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     3661 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     3721 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     3781 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     3841 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     3901 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     3961 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     4021 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     4081 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     4141 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg
     4201 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     4261 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     4321 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     4381 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     4441 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     4501 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     4561 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     4621 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     4681 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     4741 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4801 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     4861 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     4921 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     4981 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     5041 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     5101 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     5161 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     5221 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     5281 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     5341 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     5401 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     5461 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     5521 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     5581 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     5641 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     5701 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     5761 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     5821 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     5881 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5941 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     6001 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     6061 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6121 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     6181 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6241 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     6301 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6361 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6421 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     6481 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6541 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6601 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6661 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6721 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     6781 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     6841 atacgtgtcc tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct
     6901 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     6961 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     7021 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     7081 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     7141 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     7201 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     7261 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     7321 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     7381 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     7441 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     7501 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     7561 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
     7621 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     7681 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     7741 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     7801 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
     7861 gccgc
//
