LOCUS       sequence-300416-        6270 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 6270)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6270)
  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-m4046697
FEATURES             Location/Qualifiers
     source          1..6270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(316..1119)
                     /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"
     primer_bind     1470..1488
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3047..3488)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(3053..3076)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    3371..3488
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     terminator      3555..3586
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3587..3689
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3690..3758,3759..4550)
                     /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     complement(3908..3927)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4551..4742
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4766..4793
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4805..5392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5414..5443)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(5521..6270,1..233)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 cgcgttgcat ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc
       61 tctcgcgttg catttttgtt ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc
      121 gctttcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt
      181 agcgctctcg cgttgcattt ttgttctaca aaatgaagca cagatgcttc gttactcggg
      241 taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac
      301 ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc
      361 ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc
      421 aacaataata atgtcagatc ctgtagaaac cacatcatcc acggttctat actgttgacc
      481 caatgcatct cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc
      541 ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct
      601 cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga
      661 caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt
      721 caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc
      781 tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa
      841 ttttctgtcc tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac
      901 tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt
      961 gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc
     1021 acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg
     1081 atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca
     1141 cgtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact
     1201 acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc
     1261 ggagattacc gaatcaaaaa aatttcaagg aaaccgtgta ctgcagtgca ctagtggttc
     1321 ggcggccgct ttgtacagaa aaaaaagaaa aatttgaaat ataaataacg ttcttaatac
     1381 taacataact attaaaaaaa ataaataggg acctagactt caggttgtct aactccttcc
     1441 ttttcggtta gagcggatgt gggaggaggg cgtgaatgta agcgtgacat aactaattac
     1501 atgatatcga caaaggaaaa ggggcctgtt tattgcaaaa cagcggaggt aatagacgtc
     1561 tgaggtggtt gatacaagac gtcggaaccg gaattagaga aatcattcaa agccttagca
     1621 aggtgacaac aagccgcttc acggtatgaa gtagtatcca tggcacccga gaagtatttg
     1681 tacttgttgt ataaagctaa atatctgtta tattgggtta gcgggagtaa aacgtcagat
     1741 ctcaacttca agtacatttc tttgttgagc aaaaaggtac acaaggcagc ttcttcgaag
     1801 gtagaaaagc tgacaccatt aaagacaact cttctcttta gataatttga gaaaaaccaa
     1861 tagaaatgtt tagtacttat gcaaatgatg taagcgatcg ttatccagaa gggaaccaat
     1921 ggggtgaaca ttaccatcca ttgaatgtgt gccaaaaagc tgacatcgtt agtcaggtag
     1981 aatgtcaggt ataaatagat aacggagtat acaccaggca agaaagagta tactggtgta
     2041 agacacaaaa cagtgaacga catcaaaaat aatagagtgt tgaaagccac gacatgcgaa
     2101 tactccccaa aggctcttct aaatctcatg aagtagtaag ccaaacaagt aacaacgata
     2161 gcaacgattc caccggcgac aatagatgcg gaaatgtcca gcgcacctat tggttggatc
     2221 aaaggggtaa acatgttagt caatagatta acagcatcga caccgcagaa gacacctgga
     2281 agagaacggt aataatcgtt attcaacacc catctaccgc tggtagaaac acagacaccg
     2341 gcttctgatc tttcacaagt accgtggcga cagtactcgg agtcgaaggt ggtgacaacc
     2401 cgaacagaac cttcaaggta tgtgtttgga aattgaatga ttgatccatc cattaacaca
     2461 tatctagtgt ccggtcgcaa ggattcgtaa gcaacagaac cttcgaggac gtttgtgtca
     2521 taacagtatg gaactggctt accacttgca tccttgaaaa tagtgcattc agcagccaat
     2581 acacaagcag aggtggcaaa atcagtgtat tcgatcaatt tgctaggggt ataacaaata
     2641 ttgccaacag cagagaagac tcttggtaag aaatgcaaaa aatcaccatt ggtggtacgc
     2701 agtatagtcc ctggtaaacc tggcacgacg aaaccaactt ctctagtgat aacagctgca
     2761 atcaaggggc aggccttgtc gtttgtgtag gaaccacccc tttgagagaa ccaagtatca
     2821 aagtccgcgt gcttgttagc gaaacaggtg tccgtcgaag caatatctct agtaactccg
     2881 ccatcgatag ctttgtaacc aatgatctct gaggaaaagt cggtatgctt actcatcacg
     2941 tggacaggtg taattagata gaagatggct gcaacgaaca aaaatactaa agtaaccttt
     3001 atcaactgtt taagccaatt gttgacaatc ttcattatag ttttttctcc ttgacgttaa
     3061 agtatagagg tatattaaca attttttgtt gatactttta tgacatttga ataagaagta
     3121 atacaaactg aaaatgttga aagtattagt taaagtggtt atgcagcttt tccatttata
     3181 tatctgttaa tagatcaaaa atcatcgctt cgctgattaa ttaccccaga aataaggcta
     3241 aaaaactaat cgcattatca tcctatggtt gttaatttga ttcgttaatt tgaaggtttg
     3301 tggggccagg ttactgccaa tttttcctct tcataaccat aaaagctagt attgtagaat
     3361 ctttattgtt cggagcagtg cggcgcgagg cacatctgcg tttcaggaac gcgaccggtg
     3421 aagacgagga cgcacggagg agagtcttcc gtcggagggc tgtcgcccgc tcggcggctt
     3481 ctaatccgta cttcaatata gcaagcggcc gctctaactg accatttaaa tcatacctga
     3541 cctccatagc agaaagtcaa aagcctccga ccggaggctt ttgacttgat cggcacgtaa
     3601 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat
     3661 cgagattttc aggagctaag gaagctaaaa tgagtattca acatttccgt gtcgccctta
     3721 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3781 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3841 gcggtaagat ccttgagagt ttacgccccg aagaacgttt tccaatgatg agcactttta
     3901 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3961 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4021 tcacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4081 ctgcggccaa cttacttctg gcaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4141 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4201 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4261 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4321 cggataaagt tgcaggatca cttctgcgct cggccctccc ggctggctgg tttattgctg
     4381 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4441 gtaagccctc ccgcatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4501 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa gtgaccaaac
     4561 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg
     4621 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc
     4681 acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     4741 gatgagggcc caaatgtaat cacctggctc accttcgggt gggcctttct gcgttgctgg
     4801 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgatgc tcaagtcaga
     4861 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tacccctgga agctccctcg
     4921 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4981 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5041 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5101 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5161 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5221 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5281 ttacctcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5401 tttgattttc taccgaagaa aggcccaccc gtgaaggtga gccagtgagt tgattgcagt
     5461 ccagttacgc tggagtcaag cttgtaacac gcggccgctc gggaagttcc tattctctag
     5521 ctagaaagta taggaacttc agagcgcttt tgaaaaccaa aagcgctctg atgacgcact
     5581 ttcaaaaaac caaaaacgca ccggactgta acgagctact aaaatattgc gaataccgct
     5641 tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat atccctatat
     5701 aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct ctacattttt
     5761 tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac tattcataga
     5821 gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat agagacaaaa
     5881 tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta tcactttctg
     5941 ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc ctttatcttg
     6001 aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga gtcaggcttt
     6061 ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac ggacctacag
     6121 tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa aagtaatcta
     6181 agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca aaaaagaagt
     6241 atagattctt tgttggtaaa atagcgctct
//
