LOCUS       sequence-307330-        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-m4046690
FEATURES             Location/Qualifiers
     source          1..5592
                     /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"
     primer_bind     complement(1367..1385)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1736..2539
                     /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      2622..3604
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3682..3711
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3733..4320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4332..4359)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4383..4574)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4575..5366,5367..5435))
                     /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     5198..5217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5436..5538)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5539..5570)
                     /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 tgtaaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt
     1381 cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     1441 aggtccctat ttattttttt taatagttat gttagtatta agaacgttat ttatatttca
     1501 aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg
     1561 gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga
     1621 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
     1681 agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc
     1741 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
     1801 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
     1861 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
     1921 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
     1981 caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa
     2041 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
     2101 cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa
     2161 ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg
     2221 agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt
     2281 tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc
     2341 cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga
     2401 tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg caaagggaag
     2461 ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag
     2521 atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca
     2581 aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc
     2641 attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     2701 gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg
     2761 cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     2821 gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat
     2881 ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc
     2941 atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     3001 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     3061 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     3121 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     3181 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     3241 tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat
     3301 tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa
     3361 cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta
     3421 ggttatatag ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg
     3481 tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt
     3541 tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt
     3601 ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa
     3661 ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat
     3721 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3781 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag
     3841 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3901 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3961 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4021 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4081 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4141 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4201 cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4261 cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4321 aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct
     4381 catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     4441 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     4501 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     4561 tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4621 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc
     4681 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4741 ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta
     4801 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4861 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4921 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4981 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc
     5041 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5101 gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5161 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5221 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta
     5281 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5341 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5401 ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat
     5461 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     5521 cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg
     5581 gaggtcaggt at
//
