LOCUS       sequence-291436-        6427 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 6427)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6427)
  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..6427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1022..1039
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2182..2200)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2551..3354
                     /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      3437..4419
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4497..4526
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4548..5135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5147..5174)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5198..5389)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5390..6181,6182..6250))
                     /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     6013..6032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6251..6353)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6354..6385)
                     /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 atgatagcct cagagtttag ttcccttcca tcatatgcag cttttgctac
      481 tgctcaagaa gcttatgagc aggctgttgc taatggtgat tctgaagttg ttcttaaaaa
      541 gttgaagaag tctttgaatg tggctaaatc tgaatttgac cgtgatgcag ccatgcaacg
      601 taagttggaa aagatggctg atcaagctat gacccaaatg tataaacagg ctagatctga
      661 ggacaagagg gcaaaagtta ctagtgctat gcagacaatg cttttcacta tgcttagaaa
      721 gttggataat gatgcactca acaacattat caacaatgca agagatggtt gtgttccctt
      781 gaacataata cctcttacaa cagcagccaa actaatggtt gtcataccag actataacac
      841 atataaaaat acgtgtgatg gtacaacatt tacttatgca tcagcattgt gggaaatcca
      901 acaggttgta gatgcagata gtaaaattgt tcaacttagt gaaattagta tggacaattc
      961 acctaattta gcatggcctc ttattgtaac agctttaagg gccaattctg ctgtcaaatt
     1021 actcgtgccg agaggttcta aaatcgagga aggtaagcta gtcatttgga taaatggaga
     1081 taagggttac aacggtttgg cagaagtagg caagaaattt gaaaaggaca caggtatcaa
     1141 ggtgacggtt gaacatccag acaagttaga agaaaaattc cctcaagttg ctgctactgg
     1201 tgatggtccc gacattattt tctgggccca cgatcgtttt ggtggatatg ctcagtcagg
     1261 tttgctggct gagatcaccc cagacaaggc cttccaagat aaattgtacc cattcacttg
     1321 ggatgctgtc agatacaacg gtaagcttat agcatatcca attgctgttg aagccttatc
     1381 gttgatctac aataaggact tgttacctaa cccaccaaag acatgggaag aaattccagc
     1441 tttggataaa gagctaaagg ccaagggaaa aagcgctttg atgtttaact tacaagaacc
     1501 ttatttcacc tggccattaa tcgctgctga cggtggttac gcatttaaat acgaaaacgg
     1561 taagtatgac attaaggatg tcggcgttga taatgctggt gcaaaggctg gtttgacctt
     1621 cttggttgat ttaataaaga ataaacacat gaacgctgac actgactact ctatcgccga
     1681 agccgctttc aacaagggtg aaactgctat gaccataaat ggtccatggg cctggagtaa
     1741 cattgatact tctaaagtaa actatggtgt cactgttttg cctacattca aaggccaacc
     1801 atccaagccc tttgtcggtg ttctatccgc tggcatcaat gctgcttctc caaacaagga
     1861 gctagctaaa gaatttttgg aaaattactt gttaactgac gaaggtttgg aggccgttaa
     1921 caaggacaag ccattgggtg ctgtcgcttt gaaatcttac gaagaagaat tggctaagga
     1981 tcccagaatt gccgctacca tggagaacgc ccaaaagggt gaaattatgc caaacatccc
     2041 acaaatgtca gccttctggt acgcagttag gactgccgtg attaatgctg cttccggtag
     2101 acaaaccgtc gacgaagcat taaaagatgc tcaaacgtaa acaggcccct tttcctttgt
     2161 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
     2221 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
     2281 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
     2341 gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
     2401 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     2461 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     2521 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     2581 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     2641 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     2701 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     2761 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     2821 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     2881 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     2941 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     3001 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     3061 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     3121 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     3181 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     3241 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     3301 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     3361 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     3421 cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     3481 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     3541 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     3601 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     3661 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     3721 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     3781 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     3841 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     3901 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     3961 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     4021 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     4081 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     4141 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     4201 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     4261 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     4321 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt
     4381 tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg
     4441 agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct
     4501 caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt
     4561 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4621 ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga
     4681 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4741 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4801 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4861 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4921 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4981 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa
     5041 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt
     5101 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca
     5161 cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac
     5221 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     5281 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     5341 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct
     5401 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5461 tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa
     5521 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5581 ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt
     5641 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5701 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5761 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5821 tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5881 cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg
     5941 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6001 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6061 aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6121 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6181 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6241 gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     6301 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa
     6361 agcctccggt cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc
     6421 agttaga
//
