LOCUS       sequence-298267-        5028 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 5028)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5028)
  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-m4046431
FEATURES             Location/Qualifiers
     source          1..5028
                     /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             703..720
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             721..738
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(783..801)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1152..1955
                     /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      2038..3020
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3098..3127
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3149..3736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3748..3775)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3799..3990)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3991..4782,4783..4851))
                     /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     4614..4633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4852..4954)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4955..4986)
                     /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 tattcttttg ttagcgagga aacgggaact cttatagtaa actcggtgct attattcctg
      541 gctttcgtcg tttttctctt ggttacctta gcaattttga ctgccttgag gttgtgtgcg
      601 tactgctgta atatcgtcaa cgtttcattg gtcaagcctt ctttctacgt ttactccaga
      661 gtcaaaaatt tgaactcctc tcgcgtccca gatttattgg ttctcgtgcc gagaggttct
      721 caccatcatc accatcatta aacaggcccc ttttcctttg tcgatatcat gtaattagtt
      781 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
      841 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
      901 cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc gaaccactag
      961 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     1021 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     1081 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     1141 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     1201 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     1261 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     1321 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     1381 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     1441 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     1501 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     1561 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     1621 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     1681 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     1741 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     1801 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     1861 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     1921 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     1981 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaac
     2041 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     2101 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     2161 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     2221 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     2281 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     2341 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     2401 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     2461 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     2521 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     2581 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     2641 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     2701 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     2761 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     2821 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     2881 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     2941 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     3001 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaag
     3061 cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     3121 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3181 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3241 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3301 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3361 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3421 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3481 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3541 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3601 ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt atctttatag
     3661 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     3721 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     3781 attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     3841 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     3901 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     3961 gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat
     4021 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4081 tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4141 ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag
     4201 tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4261 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4321 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4381 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc
     4441 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4501 tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4561 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4621 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa
     4681 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4741 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4801 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc
     4861 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     4921 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     4981 ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
