LOCUS       sequence-291636-        5317 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 5317)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5317)
  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-m4046711
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1072..1090)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1441..2244
                     /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      2327..3309
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3387..3416
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3438..4025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4037..4064)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4088..4279)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4280..5071,5072..5140))
                     /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     4903..4922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5141..5243)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5244..5275)
                     /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 caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggctatcg ccagtgaatt ttcctcatta cctagctatg cagctttcgc
      481 tacggctcag gaggcctacg aacaagctgt agccaacggc gactctgaag ttgttcttaa
      541 gaaattgaag aagtctttga atgtcgcgaa atctgaattt gatagagatg ctgctatgca
      601 acgtaagcta gaaaagatgg ctgaccaagc aatgacccaa atgtacaaac aagccagaag
      661 tgaggacaag cgagctaagg tgacttccgc tatgcaaaca atgctgttca ctatgttgag
      721 aaagttggat aacgatgctc tgaacaatat tataaacaat gccagagatg gttgtgtccc
      781 cctcaacatt atcccattaa ccactgccgc taaattgatg gttgttattc cggattataa
      841 tacatacaaa aacacttgcg acggtaccac cttcacatac gcctctgctc tttgggaaat
      901 ccaacaggta gtcgacgctg actccaagat agttcaattg agcgaaattt ctatggataa
      961 ctcgccaaac ctagcttggc cattaatcgt cactgcgttg agagctaata gtgcggttaa
     1021 gttgcaataa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     1081 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     1141 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     1201 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     1261 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     1321 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     1381 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     1441 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     1501 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     1561 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     1621 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     1681 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     1741 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     1801 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     1861 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     1921 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     1981 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     2041 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     2101 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     2161 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     2221 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     2281 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     2341 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     2401 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     2461 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     2521 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     2581 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     2641 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     2701 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     2761 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     2821 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     2881 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     2941 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     3001 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     3061 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     3121 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     3181 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     3241 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     3301 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag
     3361 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     3421 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3481 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3541 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3601 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3661 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3721 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3781 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3841 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3901 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt
     3961 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     4021 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     4081 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     4141 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     4201 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4261 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct
     4321 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg
     4381 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4441 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa
     4501 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4561 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4621 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4681 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt
     4741 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4801 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4861 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4921 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga
     4981 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5041 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5101 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg
     5161 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     5221 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg
     5281 ctatggaggt caggtatgat ttaaatggtc agttaga
//
