LOCUS       sequence-297237-        7352 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7352)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7352)
  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-m4046160
FEATURES             Location/Qualifiers
     source          1..7352
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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             1981..1998
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3141..3159)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3510..4313
                     /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"
     terminator      5422..5451
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5473..6060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6072..6099)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6123..6314)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6315..7106,7107..7175))
                     /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     6938..6957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7176..7278)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7279..7310)
                     /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 aagattgtca acaattggct taaacagttg ataaaggtta ctttagtatt tttgttcgtt
      541 gcagccatct tctatctaat tacacctgtc cacgtgatga gtaagcatac cgacttttcc
      601 tcagagatca ttggttacaa agctatcgat ggcggagtta ctagagatat tgcttcgacg
      661 gacacctgtt tcgctaacaa gcacgcggac tttgatactt ggttctctca aaggggtggt
      721 tcctacacaa acgacaaggc ctgccccttg attgcagctg ttatcactag agaagttggt
      781 ttcgtcgtgc caggtttacc agggactata ctgcgtacca ccaatggtga ttttttgcat
      841 ttcttaccaa gagtcttctc tgctgttggc aatatttgtt atacccctag caaattgatc
      901 gaatacactg attttgccac ctctgcttgt gtattggctg ctgaatgcac tattttcaag
      961 gatgcaagtg gtaagccagt tccatactgt tatgacacaa acgtcctcga aggttctgtt
     1021 gcttacgaat ccttgcgacc ggacactaga tatgtgttaa tggatggatc aatcattcaa
     1081 tttccaaaca cataccttga aggttctgtt cgggttgtca ccaccttcga ctccgagtac
     1141 tgtcgccacg gtacttgtga aagatcagaa gccggtgtct gtgtttctac cagcggtaga
     1201 tgggtgttga ataacgatta ttaccgttct cttccaggtg tcttctgcgg tgtcgatgct
     1261 gttaatctat tgactaacat gtttacccct ttgatccaac caataggtgc gctggacatt
     1321 tccgcatcta ttgtcgccgg tggaatcgtt gctatcgttg ttacttgttt ggcttactac
     1381 ttcatgagat ttagaagagc ctttggggag tattcgcatg tcgtggcttt caacactcta
     1441 ttatttttga tgtcgttcac tgttttgtgt cttacaccag tatactcttt cttgcctggt
     1501 gtatactccg ttatctattt atacctgaca ttctacctga ctaacgatgt cagctttttg
     1561 gcacacattc aatggatggt aatgttcacc ccattggttc ccttctggat aacgatcgct
     1621 tacatcattt gcataagtac taaacatttc tattggtttt tctcaaatta tctaaagaga
     1681 agagttgtct ttaatggtgt cagcttttct accttcgaag aagctgcctt gtgtaccttt
     1741 ttgctcaaca aagaaatgta cttgaagttg agatctgacg ttttactccc gctaacccaa
     1801 tataacagat atttagcttt atacaacaag tacaaatact tctcgggtgc catggatact
     1861 acttcatacc gtgaagcggc ttgttgtcac cttgctaagg ctttgaatga tttctctaat
     1921 tccggttccg acgtcttgta tcaaccacct cagacgtcta ttacctccgc tgttttgcaa
     1981 ctcgtgccga gaggttctaa aatcgaggaa ggtaagctag tcatttggat aaatggagat
     2041 aagggttaca acggtttggc agaagtaggc aagaaatttg aaaaggacac aggtatcaag
     2101 gtgacggttg aacatccaga caagttagaa gaaaaattcc ctcaagttgc tgctactggt
     2161 gatggtcccg acattatttt ctgggcccac gatcgttttg gtggatatgc tcagtcaggt
     2221 ttgctggctg agatcacccc agacaaggcc ttccaagata aattgtaccc attcacttgg
     2281 gatgctgtca gatacaacgg taagcttata gcatatccaa ttgctgttga agccttatcg
     2341 ttgatctaca ataaggactt gttacctaac ccaccaaaga catgggaaga aattccagct
     2401 ttggataaag agctaaaggc caagggaaaa agcgctttga tgtttaactt acaagaacct
     2461 tatttcacct ggccattaat cgctgctgac ggtggttacg catttaaata cgaaaacggt
     2521 aagtatgaca ttaaggatgt cggcgttgat aatgctggtg caaaggctgg tttgaccttc
     2581 ttggttgatt taataaagaa taaacacatg aacgctgaca ctgactactc tatcgccgaa
     2641 gccgctttca acaagggtga aactgctatg accataaatg gtccatgggc ctggagtaac
     2701 attgatactt ctaaagtaaa ctatggtgtc actgttttgc ctacattcaa aggccaacca
     2761 tccaagccct ttgtcggtgt tctatccgct ggcatcaatg ctgcttctcc aaacaaggag
     2821 ctagctaaag aatttttgga aaattacttg ttaactgacg aaggtttgga ggccgttaac
     2881 aaggacaagc cattgggtgc tgtcgctttg aaatcttacg aagaagaatt ggctaaggat
     2941 cccagaattg ccgctaccat ggagaacgcc caaaagggtg aaattatgcc aaacatccca
     3001 caaatgtcag ccttctggta cgcagttagg actgccgtga ttaatgctgc ttccggtaga
     3061 caaaccgtcg acgaagcatt aaaagatgct caaacgtaaa caggcccctt ttcctttgtc
     3121 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     3181 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     3241 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     3301 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     3361 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     3421 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     3481 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     3541 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     3601 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     3661 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     3721 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     3781 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     3841 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     3901 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     3961 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     4021 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     4081 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     4141 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     4201 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     4261 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     4321 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     4381 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     4441 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     4501 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     4561 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     4621 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttaacaaagc
     4681 atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     4741 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     4801 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     4861 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     4921 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     4981 tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat
     5041 tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa
     5101 cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta
     5161 ggttatatag ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg
     5221 tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt
     5281 tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt
     5341 ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa
     5401 ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat
     5461 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5521 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag
     5581 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5641 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5701 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5761 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5821 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5881 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5941 cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6001 cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6061 aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct
     6121 catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     6181 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     6241 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     6301 tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6361 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc
     6421 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6481 ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta
     6541 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6601 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6661 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6721 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc
     6781 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6841 gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6901 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6961 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta
     7021 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7081 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7141 ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat
     7201 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     7261 cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg
     7321 gaggtcaggt atgatttaaa tggtcagtta ga
//
