LOCUS       sequence-287709-        6474 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 6474)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6474)
  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-m4046363
FEATURES             Location/Qualifiers
     source          1..6474
                     /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"
     primer_bind     complement(2229..2247)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2598..3401
                     /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      3484..4466
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4544..4573
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4595..5182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5194..5221)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5245..5436)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5437..6228,6229..6297))
                     /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     6060..6079
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6298..6400)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6401..6432)
                     /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 ggttctaaaa tcgaggaagg taagctagtc atttggataa atggagataa gggttacaac
     1141 ggtttggcag aagtaggcaa gaaatttgaa aaggacacag gtatcaaggt gacggttgaa
     1201 catccagaca agttagaaga aaaattccct caagttgctg ctactggtga tggtcccgac
     1261 attattttct gggcccacga tcgttttggt ggatatgctc agtcaggttt gctggctgag
     1321 atcaccccag acaaggcctt ccaagataaa ttgtacccat tcacttggga tgctgtcaga
     1381 tacaacggta agcttatagc atatccaatt gctgttgaag ccttatcgtt gatctacaat
     1441 aaggacttgt tacctaaccc accaaagaca tgggaagaaa ttccagcttt ggataaagag
     1501 ctaaaggcca agggaaaaag cgctttgatg tttaacttac aagaacctta tttcacctgg
     1561 ccattaatcg ctgctgacgg tggttacgca tttaaatacg aaaacggtaa gtatgacatt
     1621 aaggatgtcg gcgttgataa tgctggtgca aaggctggtt tgaccttctt ggttgattta
     1681 ataaagaata aacacatgaa cgctgacact gactactcta tcgccgaagc cgctttcaac
     1741 aagggtgaaa ctgctatgac cataaatggt ccatgggcct ggagtaacat tgatacttct
     1801 aaagtaaact atggtgtcac tgttttgcct acattcaaag gccaaccatc caagcccttt
     1861 gtcggtgttc tatccgctgg catcaatgct gcttctccaa acaaggagct agctaaagaa
     1921 tttttggaaa attacttgtt aactgacgaa ggtttggagg ccgttaacaa ggacaagcca
     1981 ttgggtgctg tcgctttgaa atcttacgaa gaagaattgg ctaaggatcc cagaattgcc
     2041 gctaccatgg agaacgccca aaagggtgaa attatgccaa acatcccaca aatgtcagcc
     2101 ttctggtacg cagttaggac tgccgtgatt aatgctgctt ccggtagaca aaccgtcgac
     2161 gaagcattaa aagatgctca aacgtaaaca ggcccctttt cctttgtcga tatcatgtaa
     2221 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     2281 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     2341 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac
     2401 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac
     2461 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     2521 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca
     2581 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     2641 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     2701 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     2761 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     2821 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg
     2881 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     2941 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     3001 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     3061 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     3121 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     3181 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc
     3241 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa
     3301 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     3361 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     3421 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     3481 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     3541 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     3601 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     3661 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     3721 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     3781 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     3841 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     3901 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     3961 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     4021 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     4081 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     4141 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     4201 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     4261 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     4321 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     4381 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag
     4441 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt
     4501 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
     4561 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4621 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4681 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4741 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4801 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4861 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4921 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4981 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5041 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct
     5101 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     5161 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     5221 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag
     5281 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     5341 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     5401 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc
     5461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5521 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5581 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg
     5641 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5701 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5761 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5821 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5881 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5941 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa
     6001 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6061 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6121 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6181 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6241 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt
     6301 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     6361 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg
     6421 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga
//
