LOCUS       sequence-297970-        5172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5172)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5172)
  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-m4046410
FEATURES             Location/Qualifiers
     source          1..5172
                     /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             841..858
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             859..882
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(927..945)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1296..2099
                     /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      2182..3164
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3242..3271
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3293..3880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3892..3919)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3943..4134)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4135..4926,4927..4995))
                     /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     4758..4777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4996..5098)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5099..5130)
                     /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 aaatttcttg ttttcttagg aatcatcaca actgtagctg catttcacca agaatgtagt
      541 ttacagtcat gtactcaaca tcaaccatat gtagttgatg acccgtgtcc tattcacttc
      601 tattctaaat ggtatattag agtaggagct agaaaatcag cacctttaat tgaattgtgc
      661 gtggatgagg ctggttctaa atcacccatt cagtacatcg atatcggtaa ttatacagtt
      721 tcctgtttac cttttacaat taattgccag gaacctaaat tgggtagtct tgtagtgcgt
      781 tgttcgttct atgaagactt tttagagtat catgacgttc gtgttgtttt agatttcatc
      841 ctcgtgccga gaggttcttg gagccatccg cagtttgaaa aataaacagg ccccttttcc
      901 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc
      961 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     1021 taatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     1081 acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg
     1141 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     1201 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     1261 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     1321 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     1381 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     1441 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     1501 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     1561 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     1621 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     1681 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     1741 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     1801 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     1861 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     1921 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg
     1981 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     2041 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     2101 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     2161 tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     2221 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2281 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     2341 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     2401 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     2461 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     2521 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     2581 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     2641 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     2701 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     2761 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     2821 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     2881 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     2941 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     3001 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     3061 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc
     3121 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     3181 tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac
     3241 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc
     3301 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3361 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc
     3421 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3481 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3541 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3601 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3661 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3721 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3781 ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg
     3841 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag
     3901 gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc
     3961 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     4021 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     4081 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca
     4141 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4201 ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc
     4261 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4321 agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat
     4381 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4441 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4501 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4561 ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt
     4621 tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac
     4681 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4741 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4801 tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc
     4861 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4921 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4981 atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     5041 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag
     5101 tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa
     5161 tggtcagtta ga
//
