LOCUS       sequence-295556-        8030 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 8030)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8030)
  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-m4134219
FEATURES             Location/Qualifiers
     source          1..8030
                     /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             980..1057
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1085..1105
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1570..1691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1581..1600)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1635..1654
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1806..1852
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1882..2018
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2063..2134
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2360..2948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2440..2459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3448..4657
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4494..4513
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4661..5689
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5716..5764
                     /note="synthetic polyadenylation signal"
     CDS             join(6030..6098,6099..6890)
                     /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(6248..6267)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6903..7039
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7184..7255
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7260..7306)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7324..7372
                     /note="synthetic polyadenylation signal"
     enhancer        7453..7756
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7758..7960
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7911..7931
                     /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 cagtcgggac ggcgaatccc
      961 ggggctgata aatccctgga aaaggagatg gaagaagaac tttattgcag tgtctgcggc
     1021 taaccggttc aaaaagatca gttcaagcgg cgccctagat tacgacattc ctacgaccgc
     1081 ctccgagaat ctctatttcc agggtgaatt tggcttggca caacatgacg aggccgtcga
     1141 taacaagttt aataaagaac agcagaacgc tttctatgag atccttcact taccaaatct
     1201 gaacgaagag caacgcaacg ctttcattca gagcctgaaa gacgatccct cgcagtctgc
     1261 aaatctcctg gccgaagcca agaagctgaa tgacgcacag gctccaaaag ttgataacaa
     1321 gtttaacaag gagcagcaga atgctttcta cgagatattg caccttccta atctgaatga
     1381 ggaacaaaga aatgcattta tccagtccct aaaagacgat ccatcacaga gtgccaacct
     1441 gcttgcggaa gccaaaaagc tcaatgatgc ccaggccccg aaagtagacg cgaaccatca
     1501 ataaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
     1561 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1621 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1681 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1741 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
     1801 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
     1861 gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     1921 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     1981 ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc
     2041 gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt
     2101 cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg
     2161 aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact
     2221 acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga
     2281 ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2341 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2401 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2461 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2521 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2581 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2641 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2701 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2761 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2821 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2881 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2941 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3001 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3061 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3121 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3181 ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     3241 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     3301 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     3361 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     3421 gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg
     3481 gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga
     3541 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     3601 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     3661 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     3721 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     3781 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     3841 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     3901 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     3961 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     4021 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     4081 gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat
     4141 gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     4201 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     4261 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     4321 gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     4381 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     4441 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     4501 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     4561 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc
     4621 cgctaaattc tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact
     4681 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     4741 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     4801 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     4861 ctttgcatcc gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     4921 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     4981 cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga
     5041 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     5101 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     5161 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     5221 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     5281 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     5341 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     5401 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat
     5461 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     5521 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     5581 tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc
     5641 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta
     5701 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     5761 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     5821 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     5881 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     5941 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     6001 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     6061 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6121 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     6181 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     6241 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6301 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     6361 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6421 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     6481 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6541 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     6601 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     6661 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6721 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6781 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6841 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     6901 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     6961 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     7021 accagaaatt tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     7081 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     7141 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     7201 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     7261 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     7321 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     7381 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     7441 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     7501 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     7561 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     7621 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     7681 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     7741 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     7801 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     7861 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     7921 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     7981 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
