LOCUS       sequence-287741-        5730 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 5730)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5730)
  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-m4046400
FEATURES             Location/Qualifiers
     source          1..5730
                     /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"
     CDS             1417..1440
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1485..1503)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1854..2657
                     /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      2740..3722
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3800..3829
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3851..4438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4450..4477)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4501..4692)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4693..5484,5485..5553))
                     /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     5316..5335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5554..5656)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5657..5688)
                     /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 ggttcttgga gccatccgca gtttgaaaaa
     1441 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     1501 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     1561 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
     1621 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
     1681 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     1741 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     1801 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     1861 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     1921 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     1981 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     2041 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     2101 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     2161 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     2221 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     2281 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     2341 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     2401 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     2461 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     2521 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     2581 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     2641 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     2701 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     2761 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2821 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     2881 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     2941 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     3001 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     3061 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     3121 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     3181 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     3241 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     3301 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     3361 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     3421 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     3481 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     3541 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     3601 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     3661 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     3721 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     3781 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     3841 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3901 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     3961 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4021 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4081 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4141 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4201 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4261 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4321 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4381 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4441 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     4501 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     4561 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     4621 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     4681 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4741 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     4801 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4861 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     4921 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4981 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5041 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5101 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     5161 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5221 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5281 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5341 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     5401 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5461 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5521 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     5581 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     5641 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     5701 ggtcaggtat gatttaaatg gtcagttaga
//
