LOCUS       sequence-295388-        5172 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 5172)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5172)
  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-m4133627
FEATURES             Location/Qualifiers
     source          1..5172
                     /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..988
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1041..1162
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1052..1071)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1106..1125
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1277..1323
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1353..1489
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1534..1605
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1831..2419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1911..1930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3165..3233,3234..4025)
                     /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(3383..3402)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4038..4174
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4326..4397
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4402..4448)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4466..4514
                     /note="synthetic polyadenylation signal"
     enhancer        4595..4898
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4900..5102
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5053..5073
                     /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 cagtcgggga aacctatacc
      961 caatccactt ctcggcttag attctaccta agctagcttg actgactgag atacagcgta
     1021 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1081 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1141 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1201 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
     1261 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1321 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
     1381 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     1441 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     1501 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     1561 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     1621 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     1681 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     1741 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     1801 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1861 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1921 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1981 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2041 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2101 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2161 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2221 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2281 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2341 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2401 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2461 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2521 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2581 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2641 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     2701 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     2761 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     2821 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     2881 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct
     2941 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     3001 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     3061 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     3121 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     3181 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3241 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     3301 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     3361 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3421 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     3481 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3541 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     3601 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3661 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3721 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     3781 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3841 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3901 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3961 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4021 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     4081 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     4141 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     4201 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     4261 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     4321 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     4381 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     4441 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     4501 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     4561 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4621 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4681 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4741 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4801 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4861 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     4921 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4981 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     5041 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     5101 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     5161 gccagcggcc gc
//
