LOCUS       sequence-291584-        6757 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 6757)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6757)
  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-m4046557
FEATURES             Location/Qualifiers
     source          1..6757
                     /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             1352..1369
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2512..2530)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2881..3684
                     /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      3767..4749
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4827..4856
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4878..5465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5477..5504)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5528..5719)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5720..6511,6512..6580))
                     /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     6343..6362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6581..6683)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6684..6715)
                     /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 actcgtgccg agaggttcta aaatcgagga
     1381 aggtaagcta gtcatttgga taaatggaga taagggttac aacggtttgg cagaagtagg
     1441 caagaaattt gaaaaggaca caggtatcaa ggtgacggtt gaacatccag acaagttaga
     1501 agaaaaattc cctcaagttg ctgctactgg tgatggtccc gacattattt tctgggccca
     1561 cgatcgtttt ggtggatatg ctcagtcagg tttgctggct gagatcaccc cagacaaggc
     1621 cttccaagat aaattgtacc cattcacttg ggatgctgtc agatacaacg gtaagcttat
     1681 agcatatcca attgctgttg aagccttatc gttgatctac aataaggact tgttacctaa
     1741 cccaccaaag acatgggaag aaattccagc tttggataaa gagctaaagg ccaagggaaa
     1801 aagcgctttg atgtttaact tacaagaacc ttatttcacc tggccattaa tcgctgctga
     1861 cggtggttac gcatttaaat acgaaaacgg taagtatgac attaaggatg tcggcgttga
     1921 taatgctggt gcaaaggctg gtttgacctt cttggttgat ttaataaaga ataaacacat
     1981 gaacgctgac actgactact ctatcgccga agccgctttc aacaagggtg aaactgctat
     2041 gaccataaat ggtccatggg cctggagtaa cattgatact tctaaagtaa actatggtgt
     2101 cactgttttg cctacattca aaggccaacc atccaagccc tttgtcggtg ttctatccgc
     2161 tggcatcaat gctgcttctc caaacaagga gctagctaaa gaatttttgg aaaattactt
     2221 gttaactgac gaaggtttgg aggccgttaa caaggacaag ccattgggtg ctgtcgcttt
     2281 gaaatcttac gaagaagaat tggctaagga tcccagaatt gccgctacca tggagaacgc
     2341 ccaaaagggt gaaattatgc caaacatccc acaaatgtca gccttctggt acgcagttag
     2401 gactgccgtg attaatgctg cttccggtag acaaaccgtc gacgaagcat taaaagatgc
     2461 tcaaacgtaa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     2521 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     2581 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     2641 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     2701 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     2761 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     2821 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     2881 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     2941 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     3001 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     3061 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     3121 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     3181 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     3241 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     3301 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     3361 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     3421 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     3481 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     3541 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     3601 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     3661 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     3721 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     3781 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     3841 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     3901 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     3961 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     4021 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     4081 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     4141 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     4201 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     4261 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     4321 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     4381 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     4441 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     4501 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     4561 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     4621 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     4681 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     4741 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag
     4801 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     4861 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4921 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4981 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5041 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5101 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5161 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5221 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5281 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5341 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt
     5401 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     5461 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     5521 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     5581 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     5641 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     5701 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct
     5761 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg
     5821 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5881 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa
     5941 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6001 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6061 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6121 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt
     6181 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6241 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6301 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6361 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga
     6421 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6481 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6541 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg
     6601 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     6661 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg
     6721 ctatggaggt caggtatgat ttaaatggtc agttaga
//
