LOCUS       sequence-298480-        6264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6264)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6264)
  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-m4046619
FEATURES             Location/Qualifiers
     source          1..6264
                     /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"
     primer_bind     complement(2019..2037)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2388..3191
                     /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      3274..4256
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4334..4363
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4385..4972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4984..5011)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5035..5226)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5227..6018,6019..6087))
                     /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     5850..5869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6088..6190)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6191..6222)
                     /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 attgttaata attggttgaa gcagttaatt aaagttacac ttgtgttcct ttttgttgct
      541 gctattttct atttaataac acctgttcat gtcatgtcta aacatactga cttttcaagt
      601 gaaatcatag gatacaaggc tattgatggt ggtgtcactc gtgacatagc atctacagat
      661 acttgttttg ctaacaaaca tgctgatttt gacacatggt ttagccagcg tggtggtagt
      721 tatactaatg acaaagcttg cccattgatt gctgcagtca taacaagaga agtgggtttt
      781 gtcgtgcctg gtttgcctgg cacgatatta cgcacaacta atggtgactt tttgcatttc
      841 ttacctagag tttttagtgc agttggtaac atctgttaca caccatcaaa acttatagag
      901 tacactgact ttgcaacatc agcttgtgtt ttggctgctg aatgtacaat ttttaaagat
      961 gcttctggta agccagtacc atattgttat gataccaatg tactagaagg ttctgttgct
     1021 tatgaaagtt tacgccctga cacacgttat gtgctcatgg atggctctat tattcaattt
     1081 cctaacacct accttgaagg ttctgttaga gtggtaacaa cttttgattc tgagtactgt
     1141 aggcacggca cttgtgaaag atcagaagct ggtgtttgtg tatctactag tggtagatgg
     1201 gtacttaaca atgattatta cagatcttta ccaggagttt tctgtggtgt agatgctgta
     1261 aatttactta ctaatatgtt tacaccacta attcaaccta ttggtgcttt ggacatatca
     1321 gcatctatag tagctggtgg tattgtagct atcgtagtaa catgccttgc ctactatttt
     1381 atgaggttta gaagagcttt tggtgaatac agtcatgtag ttgcctttaa tactttacta
     1441 ttccttatgt cattcactgt actctgttta acaccagttt actcattctt acctggtgtt
     1501 tattctgtta tttacttgta cttgacattt tatcttacta atgatgtttc ttttttagca
     1561 catattcagt ggatggttat gttcacacct ttagtacctt tctggataac aattgcttat
     1621 atcatttgta tttccacaaa gcatttctat tggttcttta gtaattacct aaagagacgt
     1681 gtagtcttta atggtgtttc ctttagtact tttgaagaag ctgcgctgtg cacctttttg
     1741 ttaaataaag aaatgtatct aaagttgcgt agtgatgtgc tattacctct tacgcaatat
     1801 aatagatact tagctcttta taataagtac aagtatttta gtggagcaat ggatacaact
     1861 agctacagag aagctgcttg ttgtcatctc gcaaaggctc tcaatgactt cagtaactca
     1921 ggttctgatg ttctttacca accaccacaa acctctatca cctcagctgt tttgtaaaca
     1981 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     2041 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     2101 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     2161 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt
     2221 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     2281 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     2341 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta
     2401 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     2461 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     2521 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     2581 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     2641 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     2701 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     2761 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     2821 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     2881 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     2941 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     3001 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt
     3061 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     3121 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     3181 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     3241 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta
     3301 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     3361 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     3421 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     3481 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     3541 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     3601 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     3661 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     3721 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     3781 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     3841 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     3901 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     3961 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     4021 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     4081 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     4141 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     4201 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag
     4261 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg
     4321 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat
     4381 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4441 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg
     4501 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4561 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4621 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4681 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4741 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4801 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4861 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4921 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4981 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag
     5041 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     5101 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     5161 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     5221 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5281 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc
     5341 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5401 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc
     5461 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5521 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5581 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5641 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt
     5701 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg
     5761 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5821 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5881 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt
     5941 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6001 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6061 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa
     6121 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     6181 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag
     6241 gtatgattta aatggtcagt taga
//
