LOCUS       sequence-292517-        5634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF3a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5634)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5634)
  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-m4046333
FEATURES             Location/Qualifiers
     source          1..5634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        45..486
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    45..162
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     457..480
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1323..1340
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1341..1364
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1409..1427)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1778..2581
                     /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      2664..3646
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3724..3753
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3775..4362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4374..4401)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4425..4616)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4617..5408,5409..5477))
                     /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     5240..5259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5478..5580)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5581..5612)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gatttaaatg gtcagttaga gcggccgctt gctatattga agtacggatt agaagccgcc
       61 gagcgggcga cagccctccg acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc
      121 gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa agattctaca
      181 atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc cacaaacctt
      241 caaattaacg aatcaaatta acaaccatag gatgataatg cgattagttt tttagcctta
      301 tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga tatataaatg
      361 gaaaagctgc ataaccactt taactaatac tttcaacatt ttcagtttgt attacttctt
      421 attcaaatgt cataaaagta tcaacaaaaa attgttaata tacctctata ctttaacgtc
      481 aaggagaaaa aactataatg gatttgttta tgagaatctt cacaattgga actgtaactt
      541 tgaagcaagg tgaaatcaag gatgctactc cttcagattt tgttcgcgct actgcaacga
      601 taccgataca agcctcactc cctttcggat ggcttattgt tggcgttgca cttcttgctg
      661 tttttcagag cgcttccaaa atcataaccc tcaaaaagag atggcaacta gcactctcca
      721 agggtgttca ctttgtttgc aacttgctgt tgttgtttgt aacagtttac tcacaccttt
      781 tgctcgttgc tgctggcctt gaagcccctt ttctctatct ttatgcttta gtctacttct
      841 tgcagagtat aaactttgta agaataataa tgaggctttg gctttgctgg aaatgccgtt
      901 ccaaaaaccc attactttat gatgccaact attttctttg ctggcatact aattgttacg
      961 actattgtat accttacaat agtgtaactt cttcaattgt cattacttca ggtgatggca
     1021 caacaagtcc tatttctgaa catgactacc agattggtgg ttatactgaa aaatgggaat
     1081 ctggagtaaa agactgtgtt gtattacaca gttacttcac ttcagactat taccagctgt
     1141 actcaactca attgagtaca gacactggtg ttgaacatgt taccttcttc atctacaata
     1201 aaattgttga tgagcctgaa gaacatgtcc aaattcacac aatcgacggt tcatccggag
     1261 ttgttaatcc agtaatggaa ccaatttatg atgaaccgac gacgactact agcgtgcctt
     1321 tgctcgtgcc gagaggttct tggagccatc cgcagtttga aaaataaaca ggcccctttt
     1381 cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat
     1441 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     1501 tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     1561 gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt
     1621 tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
     1681 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
     1741 gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga
     1801 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa
     1861 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt
     1921 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt
     1981 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact
     2041 cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg
     2101 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg
     2161 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
     2221 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
     2281 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
     2341 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
     2401 caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc
     2461 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
     2521 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta
     2581 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     2641 attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta gaacaaaaat
     2701 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     2761 aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca
     2821 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     2881 acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt
     2941 ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt
     3001 tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta
     3061 aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca
     3121 cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat
     3181 ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc
     3241 gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat
     3301 actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc
     3361 ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt
     3421 cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag
     3481 cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa
     3541 tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga
     3601 gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa
     3661 cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg caatcaactc
     3721 actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat cttcttgaga
     3781 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3841 ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga
     3901 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3961 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4021 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4081 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4141 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4201 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4261 ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat
     4321 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa
     4381 aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg
     4441 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     4501 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4561 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac
     4621 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4681 gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt
     4741 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4801 ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct
     4861 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4921 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4981 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5041 agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg
     5101 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg
     5161 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5221 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5281 attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt
     5341 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5401 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5461 aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     5521 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca
     5581 agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtat
//
