LOCUS       sequence-287635-        5683 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 5683)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5683)
  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-m4046516
FEATURES             Location/Qualifiers
     source          1..5683
                     /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..1393
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1438..1456)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1807..2610
                     /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      2693..3675
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3753..3782
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3804..4391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4403..4430)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4454..4645)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4646..5437,5438..5506))
                     /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     5269..5288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5507..5609)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5610..5641)
                     /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 agaggttctt ggagccatcc
     1381 gcagtttgaa aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc
     1441 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
     1501 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
     1561 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac
     1621 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     1681 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     1741 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     1801 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     1861 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     1921 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     1981 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     2041 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt
     2101 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     2161 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     2221 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     2281 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     2341 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     2401 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     2461 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     2521 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     2581 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     2641 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc
     2701 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     2761 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     2821 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     2881 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     2941 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     3001 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     3061 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     3121 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     3181 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     3241 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     3301 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     3361 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     3421 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     3481 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     3541 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     3601 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt
     3661 tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga
     3721 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     3781 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3841 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3901 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3961 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4021 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4081 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4141 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4201 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4261 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg
     4321 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga
     4381 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac
     4441 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     4501 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     4561 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     4621 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag
     4681 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4741 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4801 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc
     4861 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4921 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4981 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5041 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa
     5101 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5161 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5221 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5281 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct
     5341 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5401 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5461 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag
     5521 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     5581 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac
     5641 tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga
//
