LOCUS       sequence-296390-        4981 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 4981)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4981)
  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-m4046503
FEATURES             Location/Qualifiers
     source          1..4981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             656..673
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             674..691
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(736..754)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1105..1908
                     /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      1991..2973
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3051..3080
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3102..3689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3701..3728)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3752..3943)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3944..4735,4736..4804))
                     /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     4567..4586
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4805..4907)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4908..4939)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgtattctt ttgttagcga ggaaacggga actcttatag taaactcggt
      481 gctattattc ctggctttcg tcgtttttct cttggttacc ttagcaattt tgactgcctt
      541 gaggttgtgt gcgtactgct gtaatatcgt caacgtttca ttggtcaagc cttctttcta
      601 cgtttactcc agagtcaaaa atttgaactc ctctcgcgtc ccagatttat tggttctcgt
      661 gccgagaggt tctcaccatc atcaccatca ttaaacaggc cccttttcct ttgtcgatat
      721 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
      781 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
      841 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
      901 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
      961 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     1021 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     1081 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     1141 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     1201 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     1261 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     1321 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     1381 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     1441 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     1501 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     1561 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     1621 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     1681 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     1741 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     1801 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     1861 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     1921 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     1981 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     2041 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     2101 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     2161 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2221 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     2281 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     2341 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     2401 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     2461 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     2521 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     2581 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     2641 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     2701 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     2761 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     2821 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     2881 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     2941 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     3001 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     3061 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     3121 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3181 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     3241 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3301 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3361 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3421 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3481 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3541 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     3601 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     3661 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     3721 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     3781 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     3841 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     3901 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     3961 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4021 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     4081 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     4141 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4201 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4261 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4321 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4381 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4441 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     4501 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4561 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4621 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4681 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4741 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4801 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     4861 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     4921 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     4981 a
//
