LOCUS       sequence-291628-        5641 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 5641)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5641)
  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-m4046684
FEATURES             Location/Qualifiers
     source          1..5641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1396..1414)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1765..2568
                     /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      2651..3633
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3711..3740
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3762..4349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4361..4388)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4412..4603)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4604..5395,5396..5464))
                     /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     5227..5246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5465..5567)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5568..5599)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgtcgggtt tcagaaaaat ggcctttcct agtggaaagg tcgagggctg
      481 tatggtgcaa gtaacatgcg gtacgaccac tttgaacggt ttatggcttg acgatgttgt
      541 ttactgtcca cgtcatgtca tctgtacttc agaagatatg ttgaatccaa actatgaaga
      601 cctactgatt cgcaagtcta accacaattt cttagttcag gctggtaacg ttcaattgag
      661 agttataggt cacagcatgc aaaattgtgt cttgaagtta aaagttgata ccgcaaaccc
      721 gaagactcca aagtacaaat ttgtcagaat ccaacccggt caaaccttct ccgtcttggc
      781 ttgttataac gggtccccat ctggtgtata ccagtgcgct atgaggccaa acttcactat
      841 taagggttct tttttgaatg gttcttgtgg ctccgttggt tttaatatcg actacgattg
      901 cgtcagtttc tgttacatgc atcacatgga attgcctaca ggtgttcacg ccggaaccga
      961 cctagaaggg aacttctatg gtccattcgt tgatcggcaa actgctcaag ctgcgggtac
     1021 ggataccaca attactgtaa atgtgcttgc ctggttgtac gctgctgtta taaacggtga
     1081 ccgatggttt ttaaacagat tcactaccac attgaatgac tttaatctgg tcgctatgaa
     1141 atataactac gaaccactca ctcaagatca tgtcgatatt ttaggtcctt tgtctgcaca
     1201 aactggcatc gctgttttgg acatgtgtgc gtcccttaag gaactactcc aaaacggtat
     1261 gaacgggaga acgattttgg gttcagccct attggaggac gaatttaccc cattcgatgt
     1321 ggttagacag tgctctggag taaccttcca ataaacaggc cccttttcct ttgtcgatat
     1381 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
     1441 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
     1501 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
     1561 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     1621 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     1681 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     1741 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     1801 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     1861 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     1921 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     1981 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     2041 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     2101 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     2161 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     2221 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     2281 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     2341 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     2401 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     2461 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     2521 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     2581 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     2641 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     2701 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     2761 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     2821 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2881 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     2941 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     3001 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     3061 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     3121 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     3181 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     3241 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     3301 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     3361 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     3421 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     3481 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     3541 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     3601 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     3661 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     3721 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     3781 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3841 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     3901 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3961 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4021 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4081 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4141 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4201 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     4261 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     4321 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     4381 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     4441 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     4501 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     4561 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     4621 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4681 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     4741 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     4801 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4861 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4921 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4981 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5041 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5101 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     5161 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5221 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5281 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5341 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5401 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5461 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     5521 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     5581 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     5641 a
//
