LOCUS       sequence-295141-        5208 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 5208)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5208)
  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-m4133292
FEATURES             Location/Qualifiers
     source          1..5208
                     /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..1024
                     /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"
     polyA_signal    1077..1198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1088..1107)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1142..1161
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1313..1359
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1389..1525
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1570..1641
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1867..2455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1947..1966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3201..3269,3270..4061)
                     /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(3419..3438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4074..4210
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4362..4433
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4438..4484)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4502..4550
                     /note="synthetic polyadenylation signal"
     enhancer        4631..4934
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4936..5138
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5089..5109
                     /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 cagtcgaaga ggcgatggaa
      961 aaagaatttt atcgcggtga gtgcagccaa cagattcaaa aagatatcct cgtctggtgc
     1021 cctctaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
     1081 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1141 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1201 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1261 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
     1321 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
     1381 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     1441 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     1501 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
     1561 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
     1621 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
     1681 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     1741 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     1801 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1861 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1921 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1981 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2041 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2101 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2161 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2221 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2281 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2341 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2401 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2461 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2521 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2581 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2641 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     2701 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     2761 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     2821 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2881 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     2941 atcggggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     3001 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     3061 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     3121 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     3181 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     3241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3301 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3361 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     3421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3481 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     3541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3601 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     3781 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     3841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4021 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     4081 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     4141 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     4201 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     4261 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     4321 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     4381 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     4441 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     4501 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     4561 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     4621 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4681 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4741 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4801 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4861 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4921 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4981 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     5041 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     5101 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     5161 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
