LOCUS       sequence-297717-        6312 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 6312)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6312)
  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-m4046387
FEATURES             Location/Qualifiers
     source          1..6312
                     /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             1981..1998
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1999..2022
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2067..2085)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2436..3239
                     /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      3322..4304
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4382..4411
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4433..5020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5032..5059)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5083..5274)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5275..6066,6067..6135))
                     /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     5898..5917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6136..6238)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6239..6270)
                     /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 gaggttcttg gagccatccg cagtttgaaa aataaacagg ccccttttcc
     2041 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc
     2101 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     2161 taatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     2221 acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg
     2281 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     2341 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     2401 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     2461 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     2521 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     2581 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     2641 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     2701 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     2761 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     2821 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     2881 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     2941 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     3001 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     3061 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg
     3121 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     3181 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     3241 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     3301 tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     3361 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     3421 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     3481 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     3541 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     3601 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     3661 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     3721 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     3781 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     3841 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     3901 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     3961 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     4021 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     4081 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     4141 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     4201 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc
     4261 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     4321 tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac
     4381 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc
     4441 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4501 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc
     4561 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4621 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4681 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4741 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4801 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4861 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4921 ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg
     4981 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag
     5041 gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc
     5101 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     5161 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     5221 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca
     5281 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5341 ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc
     5401 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5461 agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat
     5521 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5581 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5641 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5701 ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt
     5761 tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac
     5821 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5881 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5941 tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc
     6001 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6061 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6121 atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     6181 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag
     6241 tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa
     6301 tggtcagtta ga
//
