LOCUS       sequence-297936-        6751 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 6751)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6751)
  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-m4046462
FEATURES             Location/Qualifiers
     source          1..6751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1346..1363
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2506..2524)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2875..3678
                     /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      3761..4743
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4821..4850
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4872..5459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5471..5498)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5522..5713)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5714..6505,6506..6574))
                     /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     6337..6356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6587..6677)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6678..6709)
                     /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 atgggtttta gaaaaatggc attcccatct ggtaaagttg agggttgtat
      481 ggtacaagta acttgtggta caactacact taacggtctt tggcttgatg acgtagttta
      541 ctgtccaaga catgtgatct gcacctctga agacatgctt aaccctaatt atgaagattt
      601 actcattcgt aagtctaatc ataatttctt ggtacaggct ggtaatgttc aactcagggt
      661 tattggacat tctatgcaaa attgtgtact taagcttaag gttgatacag ccaatcctaa
      721 gacacctaag tataagtttg ttcgcattca accaggacag actttttcag tgttagcttg
      781 ttacaatggt tcaccatctg gtgtttacca atgtgctatg aggcccaatt tcactattaa
      841 gggttcattc cttaatggtt catgtggtag tgttggtttt aacatagatt atgactgtgt
      901 ctctttttgt tacatgcacc atatggaatt accaactgga gttcatgctg gcacagactt
      961 agaaggtaac ttttatggac cttttgttga caggcaaaca gcacaagcag ctggtacgga
     1021 cacaactatt acagttaatg ttttagcttg gttgtacgct gctgttataa atggagacag
     1081 gtggtttctc aatcgattta ccacaactct taatgacttt aaccttgtgg ctatgaagta
     1141 caattatgaa cctctaacac aagaccatgt tgacatacta ggacctcttt ctgctcaaac
     1201 tggaattgcc gttttagata tgtgtgcttc attaaaagaa ttactgcaaa atggtatgaa
     1261 tggacgtacc atattgggta gtgctttatt agaagatgaa tttacacctt ttgatgttgt
     1321 tagacaatgc tcaggtgtta ctttcctcgt gccgagaggt tctaaaatcg aggaaggtaa
     1381 gctagtcatt tggataaatg gagataaggg ttacaacggt ttggcagaag taggcaagaa
     1441 atttgaaaag gacacaggta tcaaggtgac ggttgaacat ccagacaagt tagaagaaaa
     1501 attccctcaa gttgctgcta ctggtgatgg tcccgacatt attttctggg cccacgatcg
     1561 ttttggtgga tatgctcagt caggtttgct ggctgagatc accccagaca aggccttcca
     1621 agataaattg tacccattca cttgggatgc tgtcagatac aacggtaagc ttatagcata
     1681 tccaattgct gttgaagcct tatcgttgat ctacaataag gacttgttac ctaacccacc
     1741 aaagacatgg gaagaaattc cagctttgga taaagagcta aaggccaagg gaaaaagcgc
     1801 tttgatgttt aacttacaag aaccttattt cacctggcca ttaatcgctg ctgacggtgg
     1861 ttacgcattt aaatacgaaa acggtaagta tgacattaag gatgtcggcg ttgataatgc
     1921 tggtgcaaag gctggtttga ccttcttggt tgatttaata aagaataaac acatgaacgc
     1981 tgacactgac tactctatcg ccgaagccgc tttcaacaag ggtgaaactg ctatgaccat
     2041 aaatggtcca tgggcctgga gtaacattga tacttctaaa gtaaactatg gtgtcactgt
     2101 tttgcctaca ttcaaaggcc aaccatccaa gccctttgtc ggtgttctat ccgctggcat
     2161 caatgctgct tctccaaaca aggagctagc taaagaattt ttggaaaatt acttgttaac
     2221 tgacgaaggt ttggaggccg ttaacaagga caagccattg ggtgctgtcg ctttgaaatc
     2281 ttacgaagaa gaattggcta aggatcccag aattgccgct accatggaga acgcccaaaa
     2341 gggtgaaatt atgccaaaca tcccacaaat gtcagccttc tggtacgcag ttaggactgc
     2401 cgtgattaat gctgcttccg gtagacaaac cgtcgacgaa gcattaaaag atgctcaaac
     2461 gtaaacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     2521 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     2581 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
     2641 atttttcttt tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg
     2701 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     2761 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     2821 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     2881 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     2941 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     3001 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     3061 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     3121 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     3181 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     3241 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     3301 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     3361 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     3421 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     3481 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     3541 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     3601 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     3661 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     3721 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     3781 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     3841 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     3901 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     3961 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     4021 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     4081 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     4141 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     4201 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     4261 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     4321 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     4381 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     4441 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     4501 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     4561 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     4621 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     4681 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     4741 tagctagaga ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac
     4801 tggactgcaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc
     4861 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4921 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg
     4981 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5041 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5101 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5161 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5221 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5281 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5341 gcacgaggga gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5401 acctctgact tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5461 acgccagcaa cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc
     5521 atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     5581 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     5641 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     5701 cctgtttggt cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5761 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct
     5821 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5881 tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat
     5941 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6001 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6061 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6121 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg
     6181 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6241 tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6301 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6361 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac
     6421 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6481 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6541 gaataagggc gacacggaaa tgttgaatac tcatwytmgc ttccttagct cctgaaaatc
     6601 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     6661 ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg
     6721 aggtcaggta tgatttaaat ggtcagttag a
//
