LOCUS       sequence-297879-        6804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast 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 6804)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6804)
  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-m4046366
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1399..1416
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2559..2577)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2928..3731
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      3814..4796
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4874..4903
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4925..5512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5524..5551)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5575..5766)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5767..6558,6559..6627))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6390..6409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6628..6730)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6731..6762)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 tcgggtttca gaaaaatggc ctttcctagt ggaaaggtcg agggctgtat ggtgcaagta
      541 acatgcggta cgaccacttt gaacggttta tggcttgacg atgttgttta ctgtccacgt
      601 catgtcatct gtacttcaga agatatgttg aatccaaact atgaagacct actgattcgc
      661 aagtctaacc acaatttctt agttcaggct ggtaacgttc aattgagagt tataggtcac
      721 agcatgcaaa attgtgtctt gaagttaaaa gttgataccg caaacccgaa gactccaaag
      781 tacaaatttg tcagaatcca acccggtcaa accttctccg tcttggcttg ttataacggg
      841 tccccatctg gtgtatacca gtgcgctatg aggccaaact tcactattaa gggttctttt
      901 ttgaatggtt cttgtggctc cgttggtttt aatatcgact acgattgcgt cagtttctgt
      961 tacatgcatc acatggaatt gcctacaggt gttcacgccg gaaccgacct agaagggaac
     1021 ttctatggtc cattcgttga tcggcaaact gctcaagctg cgggtacgga taccacaatt
     1081 actgtaaatg tgcttgcctg gttgtacgct gctgttataa acggtgaccg atggttttta
     1141 aacagattca ctaccacatt gaatgacttt aatctggtcg ctatgaaata taactacgaa
     1201 ccactcactc aagatcatgt cgatatttta ggtcctttgt ctgcacaaac tggcatcgct
     1261 gttttggaca tgtgtgcgtc ccttaaggaa ctactccaaa acggtatgaa cgggagaacg
     1321 attttgggtt cagccctatt ggaggacgaa tttaccccat tcgatgtggt tagacagtgc
     1381 tctggagtaa ccttccaact cgtgccgaga ggttctaaaa tcgaggaagg taagctagtc
     1441 atttggataa atggagataa gggttacaac ggtttggcag aagtaggcaa gaaatttgaa
     1501 aaggacacag gtatcaaggt gacggttgaa catccagaca agttagaaga aaaattccct
     1561 caagttgctg ctactggtga tggtcccgac attattttct gggcccacga tcgttttggt
     1621 ggatatgctc agtcaggttt gctggctgag atcaccccag acaaggcctt ccaagataaa
     1681 ttgtacccat tcacttggga tgctgtcaga tacaacggta agcttatagc atatccaatt
     1741 gctgttgaag ccttatcgtt gatctacaat aaggacttgt tacctaaccc accaaagaca
     1801 tgggaagaaa ttccagcttt ggataaagag ctaaaggcca agggaaaaag cgctttgatg
     1861 tttaacttac aagaacctta tttcacctgg ccattaatcg ctgctgacgg tggttacgca
     1921 tttaaatacg aaaacggtaa gtatgacatt aaggatgtcg gcgttgataa tgctggtgca
     1981 aaggctggtt tgaccttctt ggttgattta ataaagaata aacacatgaa cgctgacact
     2041 gactactcta tcgccgaagc cgctttcaac aagggtgaaa ctgctatgac cataaatggt
     2101 ccatgggcct ggagtaacat tgatacttct aaagtaaact atggtgtcac tgttttgcct
     2161 acattcaaag gccaaccatc caagcccttt gtcggtgttc tatccgctgg catcaatgct
     2221 gcttctccaa acaaggagct agctaaagaa tttttggaaa attacttgtt aactgacgaa
     2281 ggtttggagg ccgttaacaa ggacaagcca ttgggtgctg tcgctttgaa atcttacgaa
     2341 gaagaattgg ctaaggatcc cagaattgcc gctaccatgg agaacgccca aaagggtgaa
     2401 attatgccaa acatcccaca aatgtcagcc ttctggtacg cagttaggac tgccgtgatt
     2461 aatgctgctt ccggtagaca aaccgtcgac gaagcattaa aagatgctca aacgtaaaca
     2521 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     2581 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     2641 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     2701 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt
     2761 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     2821 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     2881 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta
     2941 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     3001 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     3061 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     3121 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     3181 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     3241 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     3301 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     3361 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     3421 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     3481 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     3541 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt
     3601 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     3661 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     3721 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     3781 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta
     3841 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     3901 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     3961 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     4021 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     4081 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     4141 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     4201 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     4261 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     4321 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     4381 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     4441 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     4501 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     4561 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     4621 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     4681 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     4741 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag
     4801 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg
     4861 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat
     4921 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4981 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg
     5041 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5101 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5161 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5221 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5281 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5341 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5401 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5461 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5521 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag
     5581 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     5641 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     5701 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     5761 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5821 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc
     5881 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5941 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc
     6001 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6061 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6121 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6181 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt
     6241 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg
     6301 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6361 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6421 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt
     6481 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6541 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6601 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa
     6661 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     6721 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag
     6781 gtatgattta aatggtcagt taga
//
