LOCUS       sequence-287664-        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        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..786
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(831..849)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1200..2003
                     /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      2086..3068
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3146..3175
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3197..3784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3796..3823)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3847..4038)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4039..4830,4831..4899))
                     /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     4662..4681
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4900..5002)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5003..5034)
                     /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 gattataaag atcatgatgg tgattataaa gatcatgata ttgattataa agatgatgat
      781 gataaataaa caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta
      841 cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa
      901 gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat
      961 ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta
     1021 cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
     1081 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt
     1141 gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca
     1201 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc
     1261 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca
     1321 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac
     1381 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat
     1441 ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag
     1501 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg
     1561 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa
     1621 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta
     1681 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg
     1741 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga
     1801 cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg
     1861 atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg
     1921 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
     1981 gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact
     2041 cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga agcatctgtg
     2101 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     2161 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     2221 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     2281 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     2341 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca
     2401 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     2461 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     2521 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     2581 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     2641 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     2701 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     2761 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     2821 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     2881 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     2941 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt
     3001 tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga agttcctata
     3061 ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct tgactccagc
     3121 gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt cttcggtaga
     3181 aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3301 cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3361 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3421 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3481 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3541 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3601 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3661 agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt
     3721 tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3781 gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg
     3841 ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     3901 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     3961 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     4021 tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga
     4141 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4201 agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac
     4261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt
     4501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4561 atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat
     4741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4861 aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga
     4921 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     4981 ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc
     5041 tatggaggtc aggtatgatt taaatggtca gttaga
//
