LOCUS       sequence-297672-        6798 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 6798)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6798)
  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-m4046206
FEATURES             Location/Qualifiers
     source          1..6798
                     /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"
     primer_bind     complement(2553..2571)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2922..3725
                     /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      3808..4790
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4868..4897
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4919..5506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5518..5545)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5569..5760)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5761..6552,6553..6621))
                     /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     6384..6403
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6622..6724)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6725..6756)
                     /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 aaaatcgagg aaggtaagct agtcatttgg
     1441 ataaatggag ataagggtta caacggtttg gcagaagtag gcaagaaatt tgaaaaggac
     1501 acaggtatca aggtgacggt tgaacatcca gacaagttag aagaaaaatt ccctcaagtt
     1561 gctgctactg gtgatggtcc cgacattatt ttctgggccc acgatcgttt tggtggatat
     1621 gctcagtcag gtttgctggc tgagatcacc ccagacaagg ccttccaaga taaattgtac
     1681 ccattcactt gggatgctgt cagatacaac ggtaagctta tagcatatcc aattgctgtt
     1741 gaagccttat cgttgatcta caataaggac ttgttaccta acccaccaaa gacatgggaa
     1801 gaaattccag ctttggataa agagctaaag gccaagggaa aaagcgcttt gatgtttaac
     1861 ttacaagaac cttatttcac ctggccatta atcgctgctg acggtggtta cgcatttaaa
     1921 tacgaaaacg gtaagtatga cattaaggat gtcggcgttg ataatgctgg tgcaaaggct
     1981 ggtttgacct tcttggttga tttaataaag aataaacaca tgaacgctga cactgactac
     2041 tctatcgccg aagccgcttt caacaagggt gaaactgcta tgaccataaa tggtccatgg
     2101 gcctggagta acattgatac ttctaaagta aactatggtg tcactgtttt gcctacattc
     2161 aaaggccaac catccaagcc ctttgtcggt gttctatccg ctggcatcaa tgctgcttct
     2221 ccaaacaagg agctagctaa agaatttttg gaaaattact tgttaactga cgaaggtttg
     2281 gaggccgtta acaaggacaa gccattgggt gctgtcgctt tgaaatctta cgaagaagaa
     2341 ttggctaagg atcccagaat tgccgctacc atggagaacg cccaaaaggg tgaaattatg
     2401 ccaaacatcc cacaaatgtc agccttctgg tacgcagtta ggactgccgt gattaatgct
     2461 gcttccggta gacaaaccgt cgacgaagca ttaaaagatg ctcaaacgta aacaggcccc
     2521 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     2581 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     2641 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     2701 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
     2761 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     2821 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     2881 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     2941 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     3001 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     3061 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     3121 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     3181 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     3241 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     3301 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     3361 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     3421 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     3481 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     3541 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     3601 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     3661 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     3721 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     3781 tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa
     3841 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3901 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     3961 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     4021 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     4081 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     4141 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     4201 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     4261 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     4321 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     4381 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     4441 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     4501 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     4561 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     4621 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     4681 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat
     4741 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata
     4801 ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca
     4861 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     4921 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4981 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     5041 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5101 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5161 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5221 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5281 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5341 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5401 tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5461 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5521 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     5581 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     5641 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     5701 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     5761 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5821 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     5881 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5941 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     6001 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6061 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6121 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6181 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     6241 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     6301 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6361 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6421 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     6481 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6541 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6601 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     6661 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     6721 atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga
     6781 tttaaatggt cagttaga
//
