LOCUS       sequence-297581-        5592 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 5592)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5592)
  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-m4046698
FEATURES             Location/Qualifiers
     source          1..5592
                     /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"
     primer_bind     complement(1347..1365)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1716..2519
                     /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      2602..3584
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3662..3691
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3713..4300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4312..4339)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4363..4554)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4555..5346,5347..5415))
                     /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     5178..5197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5416..5518)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5519..5550)
                     /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 tataaacagg ccccttttcc
     1321 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc
     1381 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     1441 taatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     1501 acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg
     1561 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     1621 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     1681 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     1741 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     1801 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     1861 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     1921 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     1981 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     2041 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     2101 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     2161 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     2221 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     2281 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     2341 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg
     2401 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     2461 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     2521 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     2581 tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     2641 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2701 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     2761 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     2821 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     2881 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     2941 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     3001 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     3061 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     3121 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     3181 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     3241 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     3301 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     3361 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     3421 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     3481 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc
     3541 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     3601 tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac
     3661 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc
     3721 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3781 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc
     3841 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3901 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3961 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4021 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4081 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4141 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4201 ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg
     4261 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag
     4321 gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc
     4381 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     4441 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     4501 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca
     4561 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4621 ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc
     4681 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4741 agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat
     4801 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4861 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4921 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4981 ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt
     5041 tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac
     5101 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5161 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5221 tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc
     5281 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5341 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5401 atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     5461 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag
     5521 tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa
     5581 tggtcagtta ga
//
