LOCUS       sequence-298840-        5400 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 5400)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5400)
  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-m4046339
FEATURES             Location/Qualifiers
     source          1..5400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1075..1092
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1093..1110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1155..1173)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1524..2327
                     /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      2410..3392
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3470..3499
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3521..4108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4120..4147)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4171..4362)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4363..5154,5155..5223))
                     /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     4986..5005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5224..5326)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5327..5358)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gctatcgcca gtgaattttc ctcattacct agctatgcag ctttcgctac ggctcaggag
      541 gcctacgaac aagctgtagc caacggcgac tctgaagttg ttcttaagaa attgaagaag
      601 tctttgaatg tcgcgaaatc tgaatttgat agagatgctg ctatgcaacg taagctagaa
      661 aagatggctg accaagcaat gacccaaatg tacaaacaag ccagaagtga ggacaagcga
      721 gctaaggtga cttccgctat gcaaacaatg ctgttcacta tgttgagaaa gttggataac
      781 gatgctctga acaatattat aaacaatgcc agagatggtt gtgtccccct caacattatc
      841 ccattaacca ctgccgctaa attgatggtt gttattccgg attataatac atacaaaaac
      901 acttgcgacg gtaccacctt cacatacgcc tctgctcttt gggaaatcca acaggtagtc
      961 gacgctgact ccaagatagt tcaattgagc gaaatttcta tggataactc gccaaaccta
     1021 gcttggccat taatcgtcac tgcgttgaga gctaatagtg cggttaagtt gcaactcgtg
     1081 ccgagaggtt ctcaccatca tcaccatcat taaacaggcc ccttttcctt tgtcgatatc
     1141 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     1201 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     1261 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     1321 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     1381 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     1441 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     1501 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     1561 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     1621 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     1681 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     1741 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     1801 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     1861 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     1921 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     1981 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     2041 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     2101 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     2161 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     2221 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     2281 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     2341 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     2401 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     2461 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     2521 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     2581 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     2641 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     2701 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     2761 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     2821 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     2881 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     2941 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     3001 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     3061 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     3121 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     3181 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     3241 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     3301 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     3361 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     3421 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     3481 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     3541 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3601 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     3661 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3721 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3781 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3841 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3901 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3961 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     4021 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     4081 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     4141 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     4201 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     4261 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     4321 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     4381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4441 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     4501 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     4561 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4801 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4861 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     4921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5041 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5221 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     5281 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     5341 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
