LOCUS       sequence-287694-        5442 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 5442)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5442)
  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-m4046317
FEATURES             Location/Qualifiers
     source          1..5442
                     /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..1152
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1197..1215)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1566..2369
                     /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      2452..3434
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3512..3541
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3563..4150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4162..4189)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4213..4404)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4405..5196,5197..5265))
                     /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     5028..5047
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5266..5368)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5369..5400)
                     /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 ggttctgatt ataaagatca tgatggtgat tataaagatc atgatattga ttataaagat
     1141 gatgatgata aataaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     1201 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     1261 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     1321 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
     1381 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     1441 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     1501 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     1561 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     1621 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     1681 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     1741 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     1801 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     1861 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     1921 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     1981 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     2041 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     2101 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     2161 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     2221 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     2281 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     2341 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     2401 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca
     2461 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     2521 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     2581 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     2641 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     2701 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     2761 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     2821 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     2881 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     2941 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     3001 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     3061 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     3121 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     3181 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     3241 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     3301 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     3361 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     3421 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac
     3481 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc
     3541 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3601 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3661 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3721 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3781 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3841 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3901 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3961 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4021 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt
     4081 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag
     4141 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca
     4201 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     4261 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     4321 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     4381 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc
     4441 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4501 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4561 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc
     4621 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4681 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4741 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4801 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag
     4861 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4921 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4981 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5041 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc
     5101 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5161 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5221 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc
     5281 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     5341 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact
     5401 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
