LOCUS       sequence-295462-        7075 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 7075)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7075)
  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-m4133876
FEATURES             Location/Qualifiers
     source          1..7075
                     /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             947..991
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1044..1165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1055..1074)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1109..1128
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1280..1326
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1356..1492
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1537..1608
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1834..2422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1914..1933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2922..4131
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3968..3987
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4135..4734
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4135..4154)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4631..4651
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4761..4809
                     /note="synthetic polyadenylation signal"
     CDS             join(5075..5143,5144..5935)
                     /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(5293..5312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5948..6084
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6229..6300
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6305..6351)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6369..6417
                     /note="synthetic polyadenylation signal"
     enhancer        6498..6801
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6803..7005
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6956..6976
                     /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 cagtcgaaag aaaccgcggc
      961 agctaagttc gagaggcagc acatggacag ttaagctagc ttgactgact gagatacagc
     1021 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1081 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1141 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1201 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1261 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1321 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1381 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1441 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1501 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1561 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1621 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1681 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1741 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     1801 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1861 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1921 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1981 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2041 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2101 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2161 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2221 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2281 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2341 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2401 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2461 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2521 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2581 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2641 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2701 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     2761 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     2821 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     2881 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     2941 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     3001 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     3061 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     3121 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     3181 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     3241 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     3301 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     3361 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     3421 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     3481 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     3541 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     3601 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     3661 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     3721 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     3781 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     3841 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     3901 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     3961 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     4021 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     4081 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgacc
     4141 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     4201 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     4261 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     4321 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     4381 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     4441 ctggccgcgc agcaacagat ggagggcctc ctggcgccgc accggcccaa ggagcccgcg
     4501 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     4561 gtcgtgctcc ccggagtgga ggccgccgag cgcgccgggg tgcccgcctt cctggagacc
     4621 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     4681 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctaggctagt
     4741 atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
     4801 ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     4861 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     4921 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     4981 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     5041 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     5101 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5161 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     5221 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     5281 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5341 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     5401 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5461 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     5521 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5581 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     5641 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     5701 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5761 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5821 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5881 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     5941 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     6001 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     6061 tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct atcgaatctc
     6121 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     6181 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     6241 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     6301 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     6361 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     6421 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     6481 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6541 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6601 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6661 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6721 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6781 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag
     6841 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     6901 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     6961 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     7021 cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
