LOCUS       sequence-291529-        5029 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 5029)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5029)
  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-m4046561
FEATURES             Location/Qualifiers
     source          1..5029
                     /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..739
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(784..802)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1153..1956
                     /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      2039..3021
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3099..3128
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3150..3737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3749..3776)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3800..3991)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3992..4783,4784..4852))
                     /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     4615..4634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4853..4955)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4956..4987)
                     /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 tctgattata aagatcatga tggtgattat aaagatcatg atattgatta
      721 taaagatgat gatgataaat aaacaggccc cttttccttt gtcgatatca tgtaattagt
      781 tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt
      841 tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga
      901 acgttattta tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta
      961 gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
     1021 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     1081 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa
     1141 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     1201 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     1261 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     1321 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     1381 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac
     1441 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     1501 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     1561 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     1621 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     1681 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     1741 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag
     1801 tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga
     1861 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     1921 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     1981 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa
     2041 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
     2101 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
     2161 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
     2221 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
     2281 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
     2341 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
     2401 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
     2461 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
     2521 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
     2581 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
     2641 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
     2701 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
     2761 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
     2821 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
     2881 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg
     2941 gtgcgttttt ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc
     3001 tgaagttcct atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa
     3061 gcttgactcc agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc
     3121 tttcttcggt agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3181 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3241 ccaactcttt ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3301 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3361 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3421 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3481 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3541 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3601 gggtcggaac aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata
     3661 gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg
     3721 ggcggagcct atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt
     3781 gattacattt gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     3841 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     3901 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     3961 tgttaagggc ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta
     4021 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4081 ctacgatgcg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4141 gctcaccggc tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa
     4201 gtgatcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4261 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4321 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4381 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4441 ccagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4501 ttactgtcat gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat
     4561 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4621 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta
     4681 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4741 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4801 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt
     4861 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     4921 tatggtgaaa gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct
     4981 tttgactttc tgctatggag gtcaggtatg atttaaatgg tcagttaga
//
