LOCUS       sequence-295467-        7859 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 7859)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7859)
  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-m4133947
FEATURES             Location/Qualifiers
     source          1..7859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             920..940
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1289..1306
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1399..1520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1410..1429)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1464..1483
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1635..1681
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1711..1847
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1892..1963
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2189..2777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2269..2288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3277..4486
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4323..4342
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4490..5518
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5545..5593
                     /note="synthetic polyadenylation signal"
     CDS             join(5859..5927,5928..6719)
                     /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(6077..6096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6732..6868
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7013..7084
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7089..7135)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7153..7201
                     /note="synthetic polyadenylation signal"
     enhancer        7282..7585
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7587..7789
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7740..7760
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7858..7859,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggt tttagaaaaa tggcattccc atctggtaaa gttgagggtt gtatggtaca
       61 agtaacttgt ggtacaacta cacttaacgg tctttggctt gatgacgtag tttactgtcc
      121 aagacatgtg atctgcacct ctgaagacat gcttaaccct aattatgaag atttactcat
      181 tcgtaagtct aatcataatt tcttggtaca ggctggtaat gttcaactca gggttattgg
      241 acattctatg caaaattgtg tacttaagct taaggttgat acagccaatc ctaagacacc
      301 taagtataag tttgttcgca ttcaaccagg acagactttt tcagtgttag cttgttacaa
      361 tggttcacca tctggtgttt accaatgtgc tatgaggccc aatttcacta ttaagggttc
      421 attccttaat ggttcatgtg gtagtgttgg ttttaacata gattatgact gtgtctcttt
      481 ttgttacatg caccatatgg aattaccaac tggagttcat gctggcacag acttagaagg
      541 taacttttat ggaccttttg ttgacaggca aacagcacaa gcagctggta cggacacaac
      601 tattacagtt aatgttttag cttggttgta cgctgctgtt ataaatggag acaggtggtt
      661 tctcaatcga tttaccacaa ctcttaatga ctttaacctt gtggctatga agtacaatta
      721 tgaacctcta acacaagacc atgttgacat actaggacct ctttctgctc aaactggaat
      781 tgccgtttta gatatgtgtg cttcattaaa agaattactg caaaatggta tgaatggacg
      841 taccatattg ggtagtgctt tattagaaga tgaatttaca ccttttgatg ttgttagaca
      901 atgctcaggt gttactttcg aaaaccttta ctttcagtcg atgagtgaca aaatcattca
      961 tctaaccgat gactcctttg atacagacgt gttgaaggcc gatggtgcga tactggtaga
     1021 tttctgggct gaatggtgtg gcccgtgcaa gatgatcgca cccattctgg acgagatcgc
     1081 cgacgagtat caaggaaaat taactgtcgc taagcttaat atagatcaga acccagggac
     1141 ggcacctaaa tacggcatca ggggaatacc taccctcctg ctctttaaga atggggaagt
     1201 tgccgcgaca aaagtgggag ccctgtctaa gggccagcta aaagagttct tggacgctaa
     1261 cctggcaggt tcaggctcgg ggcacatgca ccatcaccat caccatagca gcgggctcgt
     1321 gccacgcggc taaggatccg caggcctctg ctagcttgac tgactgagat acagcgtacc
     1381 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1441 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1501 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1561 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
     1621 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1681 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
     1741 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
     1801 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
     1861 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
     1921 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
     1981 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
     2041 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
     2101 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
     2161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3001 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa
     3061 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc
     3121 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg
     3181 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat
     3241 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc
     3301 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga
     3361 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     3421 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     3481 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     3541 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     3601 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     3661 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     3721 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     3781 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     3841 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     3901 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc
     3961 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     4021 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     4081 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     4141 taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     4201 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     4261 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     4321 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     4381 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg
     4441 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa
     4501 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     4561 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     4621 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     4681 ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt
     4741 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4801 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc
     4861 tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4921 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4981 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     5041 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     5101 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     5161 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     5221 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cccggctccg
     5281 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     5341 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     5401 tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga
     5461 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataagc
     5521 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
     5581 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     5641 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     5701 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     5761 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     5821 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     5881 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5941 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     6001 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     6061 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6121 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     6181 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6241 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     6301 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6361 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     6421 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     6481 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6541 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6601 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6661 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6721 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     6781 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     6841 gtcctcctta ccagaaattt atccttaaga tcgtttaaac tcgactctgg ctctatcgaa
     6901 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     6961 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     7021 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     7081 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     7141 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     7201 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     7261 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     7321 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     7381 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7441 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     7501 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     7561 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     7621 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     7681 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     7741 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     7801 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
