LOCUS       sequence-298252-        5311 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 5311)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5311)
  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-m4046616
FEATURES             Location/Qualifiers
     source          1..5311
                     /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(1066..1084)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1435..2238
                     /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      2321..3303
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3381..3410
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3432..4019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4031..4058)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4082..4273)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4274..5065,5066..5134))
                     /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     4897..4916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5135..5237)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5238..5269)
                     /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 tnnnnnnncg
      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 ataaacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     1081 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     1141 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
     1201 atttttcttt tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg
     1261 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     1321 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     1381 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     1441 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     1501 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     1561 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     1621 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     1681 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     1741 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     1801 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     1861 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     1921 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     1981 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     2041 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     2101 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     2161 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     2221 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     2281 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     2341 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     2401 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     2461 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     2521 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     2581 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     2641 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     2701 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     2761 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     2821 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     2881 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     2941 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     3001 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     3061 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     3121 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     3181 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     3241 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     3301 tagctagaga ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac
     3361 tggactgcaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc
     3421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg
     3541 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3601 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3661 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3721 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3781 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3841 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3901 gcacgaggga gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3961 acctctgact tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4021 acgccagcaa cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc
     4081 atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     4141 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     4201 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     4261 cctgtttggt cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4321 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct
     4381 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4441 tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat
     4501 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4561 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4621 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4681 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg
     4741 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4801 tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4861 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4921 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac
     4981 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5041 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5101 gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc
     5161 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     5221 ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg
     5281 aggtcaggta tgatttaaat ggtcagttag a
//
