LOCUS       sequence-287718-        5766 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 5766)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5766)
  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-m4046371
FEATURES             Location/Qualifiers
     source          1..5766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1393..1410
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1411..1476
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1521..1539)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1890..2693
                     /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      2776..3758
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3836..3865
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3887..4474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4486..4513)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4537..4728)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4729..5520,5521..5589))
                     /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     5352..5371
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5590..5692)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5693..5724)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 ggttttagaa aaatggcatt cccatctggt aaagttgagg gttgtatggt acaagtaact
      541 tgtggtacaa ctacacttaa cggtctttgg cttgatgacg tagtttactg tccaagacat
      601 gtgatctgca cctctgaaga catgcttaac cctaattatg aagatttact cattcgtaag
      661 tctaatcata atttcttggt acaggctggt aatgttcaac tcagggttat tggacattct
      721 atgcaaaatt gtgtacttaa gcttaaggtt gatacagcca atcctaagac acctaagtat
      781 aagtttgttc gcattcaacc aggacagact ttttcagtgt tagcttgtta caatggttca
      841 ccatctggtg tttaccaatg tgctatgagg cccaatttca ctattaaggg ttcattcctt
      901 aatggttcat gtggtagtgt tggttttaac atagattatg actgtgtctc tttttgttac
      961 atgcaccata tggaattacc aactggagtt catgctggca cagacttaga aggtaacttt
     1021 tatggacctt ttgttgacag gcaaacagca caagcagctg gtacggacac aactattaca
     1081 gttaatgttt tagcttggtt gtacgctgct gttataaatg gagacaggtg gtttctcaat
     1141 cgatttacca caactcttaa tgactttaac cttgtggcta tgaagtacaa ttatgaacct
     1201 ctaacacaag accatgttga catactagga cctctttctg ctcaaactgg aattgccgtt
     1261 ttagatatgt gtgcttcatt aaaagaatta ctgcaaaatg gtatgaatgg acgtaccata
     1321 ttgggtagtg ctttattaga agatgaattt acaccttttg atgttgttag acaatgctca
     1381 ggtgttactt tcctcgtgcc gagaggttct gattataaag atcatgatgg tgattataaa
     1441 gatcatgata ttgattataa agatgatgat gataaataaa caggcccctt ttcctttgtc
     1501 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     1561 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     1621 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     1681 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     1741 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     1801 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     1861 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     1921 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     1981 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     2041 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     2101 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     2161 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     2221 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     2281 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     2341 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     2401 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     2461 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     2521 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     2581 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     2641 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     2701 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     2761 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     2821 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2881 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     2941 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3001 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     3061 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     3121 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     3181 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     3241 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     3301 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     3361 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3421 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     3481 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     3541 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     3601 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     3661 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     3721 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     3781 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     3841 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     3901 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3961 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     4021 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4081 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4141 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4201 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4261 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4321 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     4381 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     4441 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     4501 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     4561 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4621 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4681 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     4741 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4801 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     4861 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4921 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     4981 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5041 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5101 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5161 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5221 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     5281 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5341 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5401 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5461 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5521 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5581 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     5641 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     5701 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     5761 gttaga
//
