LOCUS       sequence-287560-        5725 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 5725)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5725)
  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-m4046283
FEATURES             Location/Qualifiers
     source          1..5725
                     /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"
     CDS             1370..1435
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1480..1498)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1849..2652
                     /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      2735..3717
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3795..3824
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3846..4433
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4445..4472)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4496..4687)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4688..5479,5480..5548))
                     /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     5311..5330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5549..5651)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5652..5683)
                     /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 agaggttctg attataaaga
     1381 tcatgatggt gattataaag atcatgatat tgattataaa gatgatgatg ataaataaac
     1441 aggccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc
     1501 tcctcccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     1561 tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     1621 cttttttttc tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct
     1681 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     1741 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     1801 ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct
     1861 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     1921 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     1981 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     2041 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     2101 aattttttac tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag
     2161 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     2221 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     2281 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     2341 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     2401 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     2461 ggtttagatg acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt
     2521 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     2581 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     2641 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     2701 gcttcaattt aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt
     2761 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     2821 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     2881 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2941 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     3001 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     3061 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     3121 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     3181 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     3241 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     3301 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     3361 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     3421 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     3481 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     3541 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     3601 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     3661 gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     3721 gagaatagga acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact
     3781 gcaatcaact cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga
     3841 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3901 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg
     3961 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4021 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4081 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4141 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4201 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4261 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4321 gggagcttcc aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4381 gacttgagca tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4441 gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga
     4501 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     4561 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4621 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4681 tggtcactta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4741 catccatagt tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat
     4801 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4861 caataaacca gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct
     4921 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4981 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5041 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5101 aaaaagcggt tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt
     5161 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat
     5221 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5281 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5341 aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt
     5401 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5461 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5521 gggcgacacg gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata
     5581 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     5641 cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca
     5701 ggtatgattt aaatggtcag ttaga
//
