LOCUS       sequence-313507-        7551 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 7551)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7551)
  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-m4133731
FEATURES             Location/Qualifiers
     source          1..7551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1504..1524
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1528..1593
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1662..1783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1673..1692)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1727..1746
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1898..1944
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1974..2110
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2155..2226
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2452..3040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2532..2551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3540..4749
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4586..4605
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4753..5781
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5808..5856
                     /note="synthetic polyadenylation signal"
     CDS             join(6122..6190,6191..6982)
                     /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(6340..6359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6995..7131
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7283..7354
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7359..7405)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7423..7471
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgagtgg gttccggaaa
      601 atggcgtttc cctccggtaa ggtcgaagga tgcatggttc aggtgacctg tggcacaact
      661 acgttaaacg gcctatggct ggacgatgtg gtatactgtc ctcgacacgt catctgcaca
      721 tcagaggaca tgctgaatcc aaactatgag gatttgctca taagaaagag caatcataac
      781 tttctggtgc aagctggaaa tgtacagctt agggttattg ggcactctat gcagaactgc
      841 gtgttaaaac tcaaggtgga taccgccaat ccgaaaacac ctaagtacaa attcgtgcgc
      901 atccagccag gccaaacctt ctctgtcctg gcatgttata acggcagccc ctcaggtgtc
      961 taccagtgcg ccatgcgccc taattttact attaagggtt ccttcttgaa cgggtcgtgc
     1021 ggatcggtcg gctttaatat cgactatgac tgtgtgagtt tttgctacat gcatcacatg
     1081 gaacttccga ctggggttca cgctggaacg gatctggagg gtaacttcta tggacccttc
     1141 gtggatagac agaccgccca ggcagccggc actgacacta ccattacggt gaatgttcta
     1201 gcttggctct acgcagctgt aataaacggc gatcgttggt tcttgaacag gtttacaaca
     1261 accctgaacg actttaatct agtcgccatg aaatataatt atgaaccact gacccaagat
     1321 catgtagaca ttctcgggcc actgagcgca cagacaggga tcgctgtgct ggacatgtgt
     1381 gccagtttaa aggagctgct tcagaatggt atgaacggcc gaacaatcct agggtctgcg
     1441 ctcttagagg atgaatttac ccccttcgat gtagtgcggc aatgtagcgg cgttactttt
     1501 caagaaaacc tttactttca gtcgatggat tataaggacc acgatggaga ctacaaagat
     1561 catgacattg attacaagga cgacgatgac aaataaggat ccgcaggcct ctgctagctt
     1621 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
     1681 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1741 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1801 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     1861 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
     1921 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
     1981 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     2041 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     2101 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
     2161 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     2221 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     2281 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     2341 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     2401 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2461 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2521 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2581 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2641 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2701 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2761 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2821 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2881 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2941 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3001 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3061 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3121 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3181 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3241 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     3301 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     3361 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     3421 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     3481 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     3541 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc
     3601 cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg
     3661 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
     3721 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
     3781 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
     3841 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
     3901 cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg
     3961 ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca
     4021 agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca
     4081 gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg
     4141 tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg
     4201 ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac
     4261 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
     4321 ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg
     4381 acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg
     4441 taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga
     4501 caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt
     4561 ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg
     4621 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata
     4681 tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt
     4741 ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     4801 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     4861 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     4921 ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct cccgattccg
     4981 gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc ccgccgtgca
     5041 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     5101 gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg gttcggccca
     5161 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     5221 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     5281 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     5341 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     5401 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     5461 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     5521 cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     5581 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     5641 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc
     5701 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     5761 cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat aaaatcgcta
     5821 tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt
     5881 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     5941 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     6001 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     6061 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     6121 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6181 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6241 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6301 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6361 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6421 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6481 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6541 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6601 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6661 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     6721 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6781 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6841 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6901 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6961 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     7021 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     7081 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     7141 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     7201 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     7261 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     7321 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     7381 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     7441 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     7501 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc g
//
