LOCUS       sequence-287589-        5401 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 5401)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5401)
  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-m4046527
FEATURES             Location/Qualifiers
     source          1..5401
                     /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             1028..1045
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1046..1111
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1156..1174)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1525..2328
                     /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      2411..3393
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3471..3500
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3522..4109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4121..4148)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4172..4363)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4364..5155,5156..5224))
                     /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     4987..5006
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5225..5327)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5328..5359)
                     /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 gttgcaactc gtgccgagag gttctgatta taaagatcat gatggtgatt ataaagatca
     1081 tgatattgat tataaagatg atgatgataa ataaacaggc cccttttcct ttgtcgatat
     1141 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
     1201 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
     1261 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
     1321 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     1381 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     1441 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     1501 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     1561 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     1621 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     1681 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     1741 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     1801 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     1861 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     1921 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     1981 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     2041 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     2101 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     2161 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     2221 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     2281 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     2341 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     2401 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     2461 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     2521 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     2581 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2641 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     2701 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     2761 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     2821 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     2881 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     2941 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     3001 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     3061 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     3121 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     3181 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     3241 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     3301 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     3361 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     3421 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     3481 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     3541 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3601 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     3661 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3721 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3781 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3841 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3901 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3961 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     4021 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     4081 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     4141 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     4201 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     4261 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     4321 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     4381 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4441 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     4501 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     4561 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4621 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4681 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4741 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4801 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4861 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     4921 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4981 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5041 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5101 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5161 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5221 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     5281 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     5341 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     5401 a
//
