LOCUS       sequence-295789-        5520 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 5520)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5520)
  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-m4133558
FEATURES             Location/Qualifiers
     source          1..5520
                     /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             1295..1312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1389..1510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1400..1419)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1454..1473
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1625..1671
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1701..1837
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1882..1953
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2179..2767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2259..2278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3513..3581,3582..4373)
                     /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(3731..3750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4386..4522
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4674..4745
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4750..4796)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4814..4862
                     /note="synthetic polyadenylation signal"
     enhancer        4943..5246
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5248..5450
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5401..5421
                     /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 cagtcgatga gtgacaaaat
      961 cattcatcta accgatgact cctttgatac agacgtgttg aaggccgatg gtgcgatact
     1021 ggtagatttc tgggctgaat ggtgtggccc gtgcaagatg atcgcaccca ttctggacga
     1081 gatcgccgac gagtatcaag gaaaattaac tgtcgctaag cttaatatag atcagaaccc
     1141 agggacggca cctaaatacg gcatcagggg aatacctacc ctcctgctct ttaagaatgg
     1201 ggaagttgcc gcgacaaaag tgggagccct gtctaagggc cagctaaaag agttcttgga
     1261 cgctaacctg gcaggttcag gctcggggca catgcaccat caccatcacc atagcagcgg
     1321 gctcgtgcca cgcggctaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
     1381 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1441 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1501 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1561 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
     1621 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
     1681 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     1741 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     1801 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     1861 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     1921 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     1981 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     2041 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     2101 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2161 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2221 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2281 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2341 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2401 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2461 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2521 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2581 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2641 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2701 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2761 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2821 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2881 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2941 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3001 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     3061 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     3121 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     3181 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     3241 aggaatatcg aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     3301 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     3361 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     3421 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     3481 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     3541 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3601 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     3661 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     3721 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3781 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     3841 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3901 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     3961 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4021 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     4081 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     4141 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4201 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4261 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4321 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     4381 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     4441 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     4501 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     4561 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     4621 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     4681 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     4741 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     4801 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     4861 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     4921 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4981 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     5041 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     5101 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     5161 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     5221 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     5281 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     5341 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     5401 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     5461 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
