LOCUS       sequence-300424-        4992 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 4992)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4992)
  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-m4046705
FEATURES             Location/Qualifiers
     source          1..4992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1012..1041
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1063..1650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1662..1689)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1713..1904)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(1905..2696,2697..2765))
                     /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     2528..2547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2766..2868)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2869..2900)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2967..3408
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2967..3084
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3379..3402
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(3689..3707)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             4058..4861
                     /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      join(4944..4992,1..934)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
       61 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
      121 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
      181 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
      241 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
      301 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
      361 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
      421 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
      481 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
      541 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
      601 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
      661 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
      721 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
      781 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc
      841 tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt
      901 ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg
      961 ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct
     1021 tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct
     1081 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1141 ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca
     1201 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1261 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1321 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1381 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1441 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1501 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc
     1561 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga
     1621 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga
     1681 aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa
     1741 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     1801 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     1861 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc
     1921 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1981 gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata
     2041 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg
     2101 gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2161 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2221 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2281 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2341 cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2401 ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac
     2461 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2521 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2581 tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2641 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2701 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     2761 ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     2821 gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct
     2881 ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta
     2941 gagcggccgc ttgctatatt gaagtacgga ttagaagccg ccgagcgggc gacagccctc
     3001 cgacggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga
     3061 tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt
     3121 atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaattaa cgaatcaaat
     3181 taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg gtaattaatc
     3241 agcgaagcga tgatttttga tctattaaca gatatataaa tggaaaagct gcataaccac
     3301 tttaactaat actttcaaca ttttcagttt gtattacttc ttattcaaat gtcataaaag
     3361 tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaactataa
     3421 tgtactcatt cgtttcggaa gagacaggta cgttaatagt taatagcgta cttctttttc
     3481 ttgctttcgt ggtattcttg ctagttacac tagccatcct tactgcgctt cgattgtgtg
     3541 cgtactgctg caatattgtt aacgtgagtc ttgtaaaacc ttctttttac gtttactctc
     3601 gtgttaaaaa tctgaattct tctagagttc ctgatcttct ggtctaaaca ggcccctttt
     3661 cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat
     3721 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     3781 tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     3841 gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt
     3901 tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
     3961 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
     4021 gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga
     4081 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa
     4141 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt
     4201 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt
     4261 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact
     4321 cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg
     4381 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg
     4441 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
     4501 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
     4561 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
     4621 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
     4681 caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc
     4741 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
     4801 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta
     4861 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     4921 attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta gaacaaaaat
     4981 gcaacgcgag ag
//
