LOCUS       sequence-317734-        7509 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 7509)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7509)
  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-m4133862
FEATURES             Location/Qualifiers
     source          1..7509
                     /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             1525..1551
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1620..1741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1631..1650)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1685..1704
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1856..1902
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1932..2068
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2113..2184
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2410..2998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2490..2509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3498..4707
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4544..4563
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4711..5739
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5766..5814
                     /note="synthetic polyadenylation signal"
     CDS             join(6080..6148,6149..6940)
                     /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(6298..6317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6953..7089
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7241..7312
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7317..7363)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7381..7429
                     /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 gtcgtacccc tatgacgtac cagattatgc gtaaggatcc
     1561 gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
     1621 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1681 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1741 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1801 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
     1861 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     1921 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1981 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     2041 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     2101 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     2161 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     2221 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     2281 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     2341 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2401 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2461 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2521 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2581 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2641 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2701 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2761 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2821 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2881 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2941 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3001 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3061 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3121 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3181 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     3241 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     3301 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     3361 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     3421 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     3481 gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     3541 cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
     3601 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     3661 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     3721 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     3781 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     3841 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     3901 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     3961 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     4021 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     4081 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     4141 aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca
     4201 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     4261 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     4321 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     4381 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     4441 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     4501 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     4561 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     4621 taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc
     4681 tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact caccgcgacg
     4741 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     4801 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     4861 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcc
     4921 gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag cctgacctat
     4981 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     5041 gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga tcttagccag
     5101 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     5161 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     5221 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     5281 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     5341 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     5401 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     5461 gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat gctccgcatt
     5521 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     5581 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     5641 gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     5701 aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta agcctagtct
     5761 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     5821 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     5881 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     5941 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     6001 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     6061 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     6121 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6181 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6241 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     6301 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6361 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     6421 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6481 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6541 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6601 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     6661 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     6721 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6781 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6841 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6901 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     6961 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     7021 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     7081 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     7141 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     7201 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     7261 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     7321 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     7381 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     7441 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     7501 ggagttccg
//
