LOCUS       sequence-297810-        5364 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 5364)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5364)
  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-m4046642
FEATURES             Location/Qualifiers
     source          1..5364
                     /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"
     primer_bind     complement(1119..1137)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1488..2291
                     /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      2374..3356
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3434..3463
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3485..4072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4084..4111)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4135..4326)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4327..5118,5119..5187))
                     /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     4950..4969
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5188..5290)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5291..5322)
                     /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 gctatcgcca gtgaattttc ctcattacct agctatgcag ctttcgctac ggctcaggag
      541 gcctacgaac aagctgtagc caacggcgac tctgaagttg ttcttaagaa attgaagaag
      601 tctttgaatg tcgcgaaatc tgaatttgat agagatgctg ctatgcaacg taagctagaa
      661 aagatggctg accaagcaat gacccaaatg tacaaacaag ccagaagtga ggacaagcga
      721 gctaaggtga cttccgctat gcaaacaatg ctgttcacta tgttgagaaa gttggataac
      781 gatgctctga acaatattat aaacaatgcc agagatggtt gtgtccccct caacattatc
      841 ccattaacca ctgccgctaa attgatggtt gttattccgg attataatac atacaaaaac
      901 acttgcgacg gtaccacctt cacatacgcc tctgctcttt gggaaatcca acaggtagtc
      961 gacgctgact ccaagatagt tcaattgagc gaaatttcta tggataactc gccaaaccta
     1021 gcttggccat taatcgtcac tgcgttgaga gctaatagtg cggttaagtt gcaataaaca
     1081 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     1141 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     1201 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     1261 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt
     1321 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     1381 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     1441 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta
     1501 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     1561 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     1621 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     1681 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     1741 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     1801 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     1861 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     1921 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     1981 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     2041 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     2101 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt
     2161 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     2221 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     2281 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     2341 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta
     2401 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     2461 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     2521 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     2581 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     2641 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     2701 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     2761 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     2821 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     2881 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     2941 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     3001 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     3061 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     3121 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     3181 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     3241 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     3301 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag
     3361 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg
     3421 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat
     3481 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3541 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg
     3601 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3661 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3721 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3781 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3841 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3901 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3961 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4021 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4081 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag
     4141 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4201 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4261 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4321 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4381 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc
     4441 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4501 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc
     4561 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4621 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4681 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4741 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt
     4801 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg
     4861 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4921 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4981 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt
     5041 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5101 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5161 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa
     5221 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     5281 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag
     5341 gtatgattta aatggtcagt taga
//
