LOCUS       sequence-298141-        5214 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 5214)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5214)
  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-m4046571
FEATURES             Location/Qualifiers
     source          1..5214
                     /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..924
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(969..987)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1338..2141
                     /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      2224..3206
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3284..3313
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3335..3922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3934..3961)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3985..4176)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4177..4968,4969..5037))
                     /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     4800..4819
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5038..5140)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5141..5172)
                     /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 gaggttctga ttataaagat catgatggtg attataaaga tcatgatatt
      901 gattataaag atgatgatga taaataaaca ggcccctttt cctttgtcga tatcatgtaa
      961 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     1021 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     1081 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac
     1141 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac
     1201 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     1261 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca
     1321 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     1381 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     1441 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     1501 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     1561 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg
     1621 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     1681 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     1741 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     1801 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     1861 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     1921 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc
     1981 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa
     2041 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     2101 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     2161 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     2221 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     2281 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     2341 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     2401 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     2461 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     2521 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     2581 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     2641 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     2701 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     2761 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     2821 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     2881 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     2941 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     3001 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     3061 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     3121 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag
     3181 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt
     3241 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
     3301 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3361 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3421 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3481 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3541 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3601 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3661 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3721 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3781 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct
     3841 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     3901 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     3961 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag
     4021 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     4081 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     4141 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc
     4201 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4261 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4321 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg
     4381 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4441 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4501 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4561 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4621 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4681 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa
     4741 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4801 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4861 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4921 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4981 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt
     5041 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     5101 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg
     5161 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga
//
