LOCUS       sequence-295780-        5685 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 5685)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5685)
  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-m4133345
FEATURES             Location/Qualifiers
     source          1..5685
                     /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    1554..1675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1565..1584)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1619..1638
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1790..1836
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1866..2002
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2047..2118
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2344..2932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2424..2443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3678..3746,3747..4538)
                     /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(3896..3915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4551..4687
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4839..4910
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4915..4961)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4979..5027
                     /note="synthetic polyadenylation signal"
     enhancer        5108..5411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5413..5615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5566..5586
                     /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 ataagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
     1561 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1621 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1681 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1741 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
     1801 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
     1861 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     1921 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     1981 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     2041 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     2101 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     2161 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     2221 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     2281 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2521 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2761 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2881 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2941 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3001 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3061 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     3181 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     3241 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     3301 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     3361 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     3421 ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     3481 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     3541 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     3601 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     3661 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3721 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3781 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3841 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     3901 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3961 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     4021 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4081 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4141 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4201 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     4261 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     4321 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4381 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4441 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4501 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     4561 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     4621 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     4681 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     4741 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     4801 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     4861 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     4921 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     4981 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     5041 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5101 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     5161 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     5221 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     5281 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     5341 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     5401 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     5461 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     5521 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     5581 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     5641 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
