LOCUS       sequence-295465-        5679 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 5679)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5679)
  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-m4134119
FEATURES             Location/Qualifiers
     source          1..5679
                     /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             974..1051
                     /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             1079..1099
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1548..1669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1559..1578)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1613..1632
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1784..1830
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1860..1996
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2041..2112
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2338..2926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2418..2437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3672..3740,3741..4532)
                     /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(3890..3909)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4545..4681
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4833..4904
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4909..4955)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4973..5021
                     /note="synthetic polyadenylation signal"
     enhancer        5102..5405
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5407..5609
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5560..5580
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5678..5679,1..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 ggacggcgaa tcccggggct
      961 gataaatccc tggaaaagga gatggaagaa gaactttatt gcagtgtctg cggctaaccg
     1021 gttcaaaaag atcagttcaa gcggcgccct agattacgac attcctacga ccgcctccga
     1081 gaatctctat ttccagggtg aatttggctt ggcacaacat gacgaggccg tcgataacaa
     1141 gtttaataaa gaacagcaga acgctttcta tgagatcctt cacttaccaa atctgaacga
     1201 agagcaacgc aacgctttca ttcagagcct gaaagacgat ccctcgcagt ctgcaaatct
     1261 cctggccgaa gccaagaagc tgaatgacgc acaggctcca aaagttgata acaagtttaa
     1321 caaggagcag cagaatgctt tctacgagat attgcacctt cctaatctga atgaggaaca
     1381 aagaaatgca tttatccagt ccctaaaaga cgatccatca cagagtgcca acctgcttgc
     1441 ggaagccaaa aagctcaatg atgcccaggc cccgaaagta gacgcgaacc atcaataagc
     1501 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
     1561 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1621 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1681 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1741 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     1801 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     1861 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     1921 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     1981 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     2041 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     2101 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     2161 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     2221 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     2281 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2521 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2701 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2761 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2821 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2881 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2941 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3001 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3061 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3121 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     3181 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     3241 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     3301 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     3361 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     3421 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     3481 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     3541 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     3601 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     3661 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3721 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3781 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3841 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3901 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3961 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4021 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4081 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4141 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4201 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     4261 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     4321 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4381 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4441 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4501 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     4561 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     4621 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     4681 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     4741 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     4801 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     4861 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     4921 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     4981 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     5041 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     5101 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     5161 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     5221 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     5281 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     5341 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     5401 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     5461 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     5521 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     5581 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     5641 cgatcctacc atccactcga cacacccgcc agcggccgc
//
