LOCUS       sequence-310875-        5076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5076)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5076)
  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-m4046455
FEATURES             Location/Qualifiers
     source          1..5076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             679..696
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             697..762
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(807..825)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1176..1979
                     /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      2062..3044
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3122..3151
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3173..3760
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3772..3799)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3823..4014)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4015..4806,4807..4875))
                     /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     4638..4657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4876..4978)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4979..5010)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 cggattagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatggaaa agctgcataa ccactttaac taatactttc aacattttca
      361 gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaact ataatgtatt cttttgttag cgaggaaacg
      481 ggaactctta tagtaaactc ggtgctatta ttcctggctt tcgtcgtttt tctcttggtt
      541 accttagcaa ttttgactgc cttgaggttg tgtgcgtact gctgtaatat cgtcaacgtt
      601 tcattggtca agccttcttt ctacgtttac tccagagtca aaaatttgaa ctcctctcgc
      661 gtcccagatt tattggttct cgtgccgaga ggttctgatt ataaagatca tgatggtgat
      721 tataaagatc atgatattga ttataaagat gatgatgata aataaacagg ccccttttcc
      781 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc
      841 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
      901 taatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
      961 acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg
     1021 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     1081 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     1141 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     1201 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     1261 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     1321 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     1381 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     1441 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     1501 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     1561 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     1621 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     1681 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     1741 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     1801 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg
     1861 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     1921 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     1981 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     2041 tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     2101 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2161 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     2221 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     2281 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     2341 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     2401 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     2461 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     2521 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     2581 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     2641 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     2701 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     2761 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     2821 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     2881 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     2941 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc
     3001 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     3061 tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac
     3121 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc
     3181 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3241 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc
     3301 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3361 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3421 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3481 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3541 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3601 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3661 ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg
     3721 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag
     3781 gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc
     3841 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     3901 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     3961 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca
     4021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4081 ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc
     4141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4201 agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat
     4261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4441 ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt
     4501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac
     4561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4681 tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc
     4741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4861 atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     4921 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag
     4981 tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa
     5041 tggtcagtta gagcggccgc ttgctatatt gaagta
//
