LOCUS       sequence-296416-        5475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 M (membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5475)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5475)
  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-m4046181
FEATURES             Location/Qualifiers
     source          1..5475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..32)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        99..540
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    99..216
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     511..534
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1218..1235
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1236..1259
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1304..1322)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1673..2476
                     /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      2559..3541
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3619..3648
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3670..4257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4269..4296)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4320..4511)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4512..5303,5304..5372))
                     /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     5135..5154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5373..5475)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtatgattta
       61 aatggtcagt tagagcggcc gcttgctata ttgaagtacg gattagaagc cgccgagcgg
      121 gcgacagccc tccgacggaa gactctcctc cgtgcgtcct cgtcttcacc ggtcgcgttc
      181 ctgaaacgca gatgtgcctc gcgccgcact gctccgaaca ataaagattc tacaatacta
      241 gcttttatgg ttatgaagag gaaaaattgg cagtaacctg gccccacaaa ccttcaaatt
      301 aacgaatcaa attaacaacc ataggatgat aatgcgatta gttttttagc cttatttctg
      361 gggtaattaa tcagcgaagc gatgattttt gatctattaa cagatatata aatggaaaag
      421 ctgcataacc actttaacta atactttcaa cattttcagt ttgtattact tcttattcaa
      481 atgtcataaa agtatcaaca aaaaattgtt aatatacctc tatactttaa cgtcaaggag
      541 aaaaaactat aatggctgat tccaatggca ccatcacagt agaagagctg aaaaagctcc
      601 ttgaacaatg gaacttggtt ataggttttt tattcttgac ttggatttgc ctattgcagt
      661 tcgcctatgc aaacagaaac cgttttttgt acattatcaa gttaattttc ttgtggctgt
      721 tatggcctgt cactcttgct tgttttgttt tggctgccgt ctaccggata aattggatca
      781 ccggtggtat cgctattgca atggcttgtt tggttggatt gatgtggttg agttatttca
      841 ttgcctcatt cagattattc gctaggacta gatcgatgtg gtcctttaat ccagaaacga
      901 acatcttgtt gaacgtgcca ctccatggta ctattttgac cagaccgcta ttagaatctg
      961 agttggttat cggtgctgtc attctaagag ggcacttacg tatcgcgggt caccatttgg
     1021 gaagatgtga cattaaggat cttccaaaag aaatcacagt cgctacctct cgaactctat
     1081 cttactacaa gttgggtgcc tcacaaaggg ttgctggtga cagcggtttc gctgcctatt
     1141 ctcgctaccg tatcggcaac tacaaattaa ataccgatca ctcctctagt tctgacaata
     1201 ttgccttatt ggtgcaactc gtgccgagag gttcttggag ccatccgcag tttgaaaaat
     1261 aaacaggccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac
     1321 gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     1381 tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat
     1441 ttttcttttt tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt
     1501 tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca
     1561 cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt
     1621 attcttaacc caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa
     1681 agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa
     1741 tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga
     1801 attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga
     1861 tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa
     1921 gtacaatttt ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt
     1981 gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg
     2041 tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga
     2101 acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga
     2161 atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat
     2221 tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg
     2281 tgtgggttta gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt
     2341 ggtttctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga
     2401 tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg
     2461 cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat
     2521 tagagcttca atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt
     2581 ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     2641 tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt
     2701 catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     2761 ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta
     2821 tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc
     2881 ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc
     2941 actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa
     3001 aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt
     3061 tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga
     3121 acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct
     3181 attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca
     3241 ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat
     3301 aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt
     3361 tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg
     3421 aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga
     3481 aagtgcgtca tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta
     3541 gctagagaat aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg
     3601 gactgcaatc aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa
     3661 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3721 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta
     3781 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3841 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3901 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3961 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4021 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4081 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4141 acgagggagc ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4201 ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4261 gccagcaacg cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat
     4321 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     4381 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     4441 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     4501 tgtttggtca cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4561 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta
     4621 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4681 tcagcaataa accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc
     4741 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4801 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4861 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4921 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca
     4981 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg
     5041 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5101 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5161 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg
     5221 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5281 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5341 ataagggcga cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc
     5401 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     5461 cttacgtgcc gatca
//
