LOCUS       sequence-292515-        4987 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 4987)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4987)
  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-m4046262
FEATURES             Location/Qualifiers
     source          1..4987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..436
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             675..692
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             693..716
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(761..779)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1130..1933
                     /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      2016..2998
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3076..3105
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3127..3714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3726..3753)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3777..3968)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3969..4760,4761..4829))
                     /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     4592..4611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4830..4932)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4933..4964)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 atttaaatgg tcagttagag cggccgcccc acacaccata gcttcaaaat gtttctactc
       61 cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc
      121 caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta attacccgta
      181 ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc
      241 aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat ttttttctct
      301 ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag
      361 tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat
      421 agcaatctaa tctaagtttt aattacaaaa tgtactcatt cgtttcggaa gagacaggta
      481 cgttaatagt taatagcgta cttctttttc ttgctttcgt ggtattcttg ctagttacac
      541 tagccatcct tactgcgctt cgattgtgtg cgtactgctg caatattgtt aacgtgagtc
      601 ttgtaaaacc ttctttttac gtttactctc gtgttaaaaa tctgaattct tctagagttc
      661 ctgatcttct ggtcctcgtg ccgagaggtt cttggagcca tccgcagttt gaaaaataaa
      721 caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc
      781 ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
      841 ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
      901 tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc
      961 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     1021 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     1081 cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc
     1141 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
     1201 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
     1261 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
     1321 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
     1381 caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca
     1441 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
     1501 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
     1561 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
     1621 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
     1681 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1741 gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1801 ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1861 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1921 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1981 agcttcaatt taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg
     2041 tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     2101 ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat
     2161 ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     2221 catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac
     2281 ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta
     2341 gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact
     2401 gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa
     2461 agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca
     2521 agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca
     2581 gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt
     2641 ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc
     2701 tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa
     2761 aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat
     2821 atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag
     2881 cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag
     2941 tgcgtcatca gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct
     3001 agagaatagg aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac
     3061 tgcaatcaac tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg
     3121 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3181 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact
     3241 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3301 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3361 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3421 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3481 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3541 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3601 agggagcttc caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3661 tgacttgagc atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3721 agcaacgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag
     3781 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3841 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     3901 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     3961 ttggtcactt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4021 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca
     4081 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4141 gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc
     4201 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4261 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4321 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4381 aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg
     4441 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga
     4501 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4561 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4621 aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg
     4681 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4741 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4801 agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat
     4861 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     4921 acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc
     4981 aggtatg
//
