LOCUS       sequence-425637-        7545 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 7545)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7545)
  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-m4133949
FEATURES             Location/Qualifiers
     source          1..7545
                     /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"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1498..1518
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1522..1587
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1656..1777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1667..1686)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1721..1740
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1892..1938
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1968..2104
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2149..2220
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2446..3034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2526..2545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3534..4743
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4580..4599
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4747..5775
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5802..5850
                     /note="synthetic polyadenylation signal"
     CDS             join(6116..6184,6185..6976)
                     /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(6334..6353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6989..7125
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7277..7348
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7353..7399)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7417..7465
                     /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 ccatgggttt tagaaaaatg
      601 gcattcccat ctggtaaagt tgagggttgt atggtacaag taacttgtgg tacaactaca
      661 cttaacggtc tttggcttga tgacgtagtt tactgtccaa gacatgtgat ctgcacctct
      721 gaagacatgc ttaaccctaa ttatgaagat ttactcattc gtaagtctaa tcataatttc
      781 ttggtacagg ctggtaatgt tcaactcagg gttattggac attctatgca aaattgtgta
      841 cttaagctta aggttgatac agccaatcct aagacaccta agtataagtt tgttcgcatt
      901 caaccaggac agactttttc agtgttagct tgttacaatg gttcaccatc tggtgtttac
      961 caatgtgcta tgaggcccaa tttcactatt aagggttcat tccttaatgg ttcatgtggt
     1021 agtgttggtt ttaacataga ttatgactgt gtctcttttt gttacatgca ccatatggaa
     1081 ttaccaactg gagttcatgc tggcacagac ttagaaggta acttttatgg accttttgtt
     1141 gacaggcaaa cagcacaagc agctggtacg gacacaacta ttacagttaa tgttttagct
     1201 tggttgtacg ctgctgttat aaatggagac aggtggtttc tcaatcgatt taccacaact
     1261 cttaatgact ttaaccttgt ggctatgaag tacaattatg aacctctaac acaagaccat
     1321 gttgacatac taggacctct ttctgctcaa actggaattg ccgttttaga tatgtgtgct
     1381 tcattaaaag aattactgca aaatggtatg aatggacgta ccatattggg tagtgcttta
     1441 ttagaagatg aatttacacc ttttgatgtt gttagacaat gctcaggtgt tactttcgaa
     1501 aacctttact ttcagtcgat ggattataag gaccacgatg gagactacaa agatcatgac
     1561 attgattaca aggacgacga tgacaaataa ggatccgcag gcctctgcta gcttgactga
     1621 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
     1681 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1741 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1801 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     1861 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
     1921 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
     1981 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     2041 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     2101 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     2161 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     2221 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     2281 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     2341 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     2401 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2461 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2521 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2581 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2641 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2701 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2761 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2821 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2881 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2941 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3001 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3061 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3121 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3181 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3241 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     3301 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     3361 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     3421 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     3481 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc
     3541 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
     3601 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     3661 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     3721 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     3781 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     3841 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     3901 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     3961 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     4021 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     4081 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     4141 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
     4201 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
     4261 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     4321 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     4381 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
     4441 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     4501 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     4561 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     4621 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     4681 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     4741 gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     4801 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     4861 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     4921 tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat tccggaagtg
     4981 cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     5041 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     5101 gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg cccattcgga
     5161 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     5221 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     5281 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     5341 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     5401 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     5461 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     5521 ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     5581 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     5641 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc cgccgtctgg
     5701 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     5761 agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     5821 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     5881 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     5941 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     6001 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     6061 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     6121 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6181 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     6241 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6301 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6361 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6421 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6481 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     6541 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6601 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6661 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     6721 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6781 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6841 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6901 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6961 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     7021 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     7081 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     7141 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     7201 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     7261 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     7321 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     7381 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     7441 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     7501 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
