LOCUS       sequence-295394-        7066 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 7066)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7066)
  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-m4134047
FEATURES             Location/Qualifiers
     source          1..7066
                     /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"
     promoter        2913..4122
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3959..3978
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4126..4725
                     /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(4126..4145)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4622..4642
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4752..4800
                     /note="synthetic polyadenylation signal"
     CDS             join(5066..5134,5135..5926)
                     /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(5284..5303)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5939..6075
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6220..6291
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6296..6342)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6360..6408
                     /note="synthetic polyadenylation signal"
     enhancer        6489..6792
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6794..6996
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6947..6967
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7065..7066,1..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 ctggcctccg cgccgggttt tggcgcctcc
     2941 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     3001 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     3061 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     3121 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     3181 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     3241 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     3301 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     3361 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     3421 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     3481 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     3541 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     3601 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     3661 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     3721 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     3781 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     3841 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     3901 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     3961 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     4021 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     4081 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag
     4141 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4201 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4261 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4321 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4381 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4441 cagcaacaga tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4501 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4561 cccggagtgg aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4621 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4681 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc
     4741 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
     4801 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     4861 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     4921 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     4981 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     5041 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     5101 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5161 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     5221 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     5281 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5341 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     5401 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5461 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     5521 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5581 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     5641 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     5701 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5761 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5821 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5881 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     5941 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     6001 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     6061 gaaatttatc cttaagatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     6121 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     6181 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     6241 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     6301 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     6361 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
     6421 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg
     6481 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6541 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6601 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6661 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6721 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6781 gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     6841 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     6901 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     6961 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga
     7021 tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgc
//
