LOCUS       sequence-287696-        5394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5394)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5394)
  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-m4046391
FEATURES             Location/Qualifiers
     source          1..5394
                     /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             1069..1086
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1087..1104
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1149..1167)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1518..2321
                     /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      2404..3386
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3464..3493
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3515..4102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4114..4141)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4165..4356)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4357..5148,5149..5217))
                     /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     4980..4999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5218..5320)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5321..5352)
                     /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 atagcctcag agtttagttc ccttccatca tatgcagctt ttgctactgc tcaagaagct
      541 tatgagcagg ctgttgctaa tggtgattct gaagttgttc ttaaaaagtt gaagaagtct
      601 ttgaatgtgg ctaaatctga atttgaccgt gatgcagcca tgcaacgtaa gttggaaaag
      661 atggctgatc aagctatgac ccaaatgtat aaacaggcta gatctgagga caagagggca
      721 aaagttacta gtgctatgca gacaatgctt ttcactatgc ttagaaagtt ggataatgat
      781 gcactcaaca acattatcaa caatgcaaga gatggttgtg ttcccttgaa cataatacct
      841 cttacaacag cagccaaact aatggttgtc ataccagact ataacacata taaaaatacg
      901 tgtgatggta caacatttac ttatgcatca gcattgtggg aaatccaaca ggttgtagat
      961 gcagatagta aaattgttca acttagtgaa attagtatgg acaattcacc taatttagca
     1021 tggcctctta ttgtaacagc tttaagggcc aattctgctg tcaaattact cgtgccgaga
     1081 ggttctcacc atcatcacca tcattaaaca ggcccctttt cctttgtcga tatcatgtaa
     1141 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     1201 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     1261 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac
     1321 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac
     1381 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     1441 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca
     1501 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     1561 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     1621 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     1681 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     1741 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg
     1801 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     1861 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     1921 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     1981 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     2041 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     2101 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc
     2161 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa
     2221 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     2281 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     2341 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     2401 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     2461 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     2521 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     2581 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     2641 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     2701 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     2761 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     2821 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     2881 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     2941 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     3001 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     3061 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     3121 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     3181 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     3241 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     3301 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag
     3361 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt
     3421 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
     3481 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3601 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3661 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct
     4021 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     4081 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     4141 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag
     4201 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     4261 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     4321 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc
     4381 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4441 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4501 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg
     4561 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4621 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4681 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4741 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4801 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4861 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa
     4921 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4981 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5041 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5101 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5161 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt
     5221 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     5281 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg
     5341 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga
//
