LOCUS       sequence-287732-        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-m4046373
FEATURES             Location/Qualifiers
     source          1..5634
                     /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             1303..1320
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1321..1344
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1389..1407)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1758..2561
                     /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      2644..3626
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3704..3733
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3755..4342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4354..4381)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4405..4596)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4597..5388,5389..5457))
                     /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     5220..5239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5458..5560)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5561..5592)
                     /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 gacttattta tgagaatatt cacgatcggc acagtgaccc taaagcaggg tgagattaaa
      541 gatgctactc catcagattt cgttagggca accgctacta tccccattca agccagcttg
      601 ccttttggtt ggttgattgt aggagtcgct ctcttggctg ttttccaatc cgccagtaag
      661 atcattactt taaaaaagag atggcaattg gcgctttcta agggtgttca cttcgtctgt
      721 aatctgttgt tattgtttgt cactgtttat tcccatttgc tcctagttgc cgctggttta
      781 gaagctccat tcctgtactt gtacgctttg gtctatttcc tgcaatctat caactttgtt
      841 cgtatcatta tgagattgtg gttgtgctgg aaatgtcgtt ctaagaaccc gttgttatac
      901 gacgctaatt acttcctttg ttggcacacc aactgttacg attattgtat tccatataac
      961 tcggttactt cctctatagt tatcacctcc ggggacggta ccacaagtcc aatttcagaa
     1021 catgactacc aaattggtgg ttacactgaa aagtgggagt ctggtgtcaa agattgcgta
     1081 gtgttacaca gctattttac ttctgattac taccagttgt atagcaccca attgtccact
     1141 gacaccggag tggaacacgt cacatttttc atctacaata agatagtcga tgaacctgaa
     1201 gaacatgttc aaatacacac tattgacggc tctagcggtg tagttaaccc agttatggaa
     1261 ccaatctatg atgagccaac aacaacgacc tctgttccct tactcgtgcc gagaggttct
     1321 tggagccatc cgcagtttga aaaataaaca ggcccctttt cctttgtcga tatcatgtaa
     1381 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     1441 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     1501 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac
     1561 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac
     1621 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     1681 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca
     1741 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     1801 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     1861 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     1921 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     1981 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg
     2041 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     2101 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     2161 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     2221 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     2281 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     2341 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc
     2401 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa
     2461 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     2521 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     2581 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     2641 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     2701 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     2761 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     2821 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     2881 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     2941 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     3001 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     3061 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     3121 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     3181 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     3241 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     3301 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     3361 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     3421 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     3481 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     3541 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag
     3601 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt
     3661 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
     3721 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3781 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3841 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3901 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3961 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4021 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4081 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4141 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4201 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct
     4261 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     4321 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     4381 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag
     4441 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     4501 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     4561 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc
     4621 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4681 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4741 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg
     4801 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4861 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4921 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4981 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5041 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5101 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa
     5161 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5221 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5281 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5341 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5401 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt
     5461 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     5521 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg
     5581 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga
//
