LOCUS       sequence-297241-        6404 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6404)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6404)
  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-m4046495
FEATURES             Location/Qualifiers
     source          1..6404
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             999..1016
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2159..2177)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2528..3331
                     /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      3414..4396
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4474..4503
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4525..5112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5124..5151)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5175..5366)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5367..6158,6159..6227))
                     /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     5990..6009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6228..6330)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6331..6362)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaatg aaaatttttt
      241 tttttgattt ttttctcttt cgatgacctc ccattgatat ttaagttaat aaacggtctt
      301 caatttctca agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgc
      361 tcattagaaa gaaagcatag caatctaatc taagttttaa ttacaaaatg atagcctcag
      421 agtttagttc ccttccatca tatgcagctt ttgctactgc tcaagaagct tatgagcagg
      481 ctgttgctaa tggtgattct gaagttgttc ttaaaaagtt gaagaagtct ttgaatgtgg
      541 ctaaatctga atttgaccgt gatgcagcca tgcaacgtaa gttggaaaag atggctgatc
      601 aagctatgac ccaaatgtat aaacaggcta gatctgagga caagagggca aaagttacta
      661 gtgctatgca gacaatgctt ttcactatgc ttagaaagtt ggataatgat gcactcaaca
      721 acattatcaa caatgcaaga gatggttgtg ttcccttgaa cataatacct cttacaacag
      781 cagccaaact aatggttgtc ataccagact ataacacata taaaaatacg tgtgatggta
      841 caacatttac ttatgcatca gcattgtggg aaatccaaca ggttgtagat gcagatagta
      901 aaattgttca acttagtgaa attagtatgg acaattcacc taatttagca tggcctctta
      961 ttgtaacagc tttaagggcc aattctgctg tcaaattact cgtgccgaga ggttctaaaa
     1021 tcgaggaagg taagctagtc atttggataa atggagataa gggttacaac ggtttggcag
     1081 aagtaggcaa gaaatttgaa aaggacacag gtatcaaggt gacggttgaa catccagaca
     1141 agttagaaga aaaattccct caagttgctg ctactggtga tggtcccgac attattttct
     1201 gggcccacga tcgttttggt ggatatgctc agtcaggttt gctggctgag atcaccccag
     1261 acaaggcctt ccaagataaa ttgtacccat tcacttggga tgctgtcaga tacaacggta
     1321 agcttatagc atatccaatt gctgttgaag ccttatcgtt gatctacaat aaggacttgt
     1381 tacctaaccc accaaagaca tgggaagaaa ttccagcttt ggataaagag ctaaaggcca
     1441 agggaaaaag cgctttgatg tttaacttac aagaacctta tttcacctgg ccattaatcg
     1501 ctgctgacgg tggttacgca tttaaatacg aaaacggtaa gtatgacatt aaggatgtcg
     1561 gcgttgataa tgctggtgca aaggctggtt tgaccttctt ggttgattta ataaagaata
     1621 aacacatgaa cgctgacact gactactcta tcgccgaagc cgctttcaac aagggtgaaa
     1681 ctgctatgac cataaatggt ccatgggcct ggagtaacat tgatacttct aaagtaaact
     1741 atggtgtcac tgttttgcct acattcaaag gccaaccatc caagcccttt gtcggtgttc
     1801 tatccgctgg catcaatgct gcttctccaa acaaggagct agctaaagaa tttttggaaa
     1861 attacttgtt aactgacgaa ggtttggagg ccgttaacaa ggacaagcca ttgggtgctg
     1921 tcgctttgaa atcttacgaa gaagaattgg ctaaggatcc cagaattgcc gctaccatgg
     1981 agaacgccca aaagggtgaa attatgccaa acatcccaca aatgtcagcc ttctggtacg
     2041 cagttaggac tgccgtgatt aatgctgctt ccggtagaca aaccgtcgac gaagcattaa
     2101 aagatgctca aacgtaaaca ggcccctttt cctttgtcga tatcatgtaa ttagttatgt
     2161 cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa ggagttagac
     2221 aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat taagaacgtt
     2281 atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac cactagtgca
     2341 ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga
     2401 acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac
     2461 atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca ggaaacgaag
     2521 ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc
     2581 tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt
     2641 tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact
     2701 aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa
     2761 ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg ctgacattgg
     2821 taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat
     2881 tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga
     2941 agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc
     3001 cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt
     3061 tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt
     3121 gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc aacagtatag
     3181 aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa gaggactatt
     3241 tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc
     3301 atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat
     3361 actaaactca caaattagag cttcaattta attatatcag ttattacccg agtaacgaag
     3421 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     3481 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     3541 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     3601 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     3661 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     3721 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     3781 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     3841 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     3901 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     3961 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     4021 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     4081 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     4141 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     4201 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     4261 gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
     4321 tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag
     4381 ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt tacaagcttg
     4441 actccagcgt aactggactg caatcaactc actggctcac cttcacgggt gggcctttct
     4501 tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4561 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4621 tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4681 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4741 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4801 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4861 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4921 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4981 ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct ttatagtcct
     5041 gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc aggggggcgg
     5101 agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc caggtgatta
     5161 catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     5221 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     5281 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     5341 agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc acctatctca
     5401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5461 atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5521 ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg cagaagtgat
     5581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgccaga
     5821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5881 gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgtaaactc
     6061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt agcttcctta
     6241 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     6301 tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg aggcttttga
     6361 ctttctgcta tggaggtcag gtatgattta aatggtcagt taga
//
