LOCUS       sequence-297876-        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-m4046470
FEATURES             Location/Qualifiers
     source          1..5475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1144..1161
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1162..1185
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1230..1248)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1599..2402
                     /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      2485..3467
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3545..3574
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3596..4183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4195..4222)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4246..4437)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4438..5229,5230..5298))
                     /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     5061..5080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5299..5401)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5402..5433)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gcagattcca acggtactat taccgttgaa gagcttaaaa agctccttga acaatggaac
      541 ctagtaatag gtttcctatt ccttacatgg atttgtcttc tacaatttgc ctatgccaac
      601 aggaataggt ttttgtatat aattaagtta attttcctct ggctgttatg gccagtaact
      661 ttagcttgtt ttgtgcttgc tgctgtttac agaataaatt ggatcaccgg tggaattgct
      721 atcgcaatgg cttgtcttgt aggcttgatg tggctcagct acttcattgc ttctttcaga
      781 ctgtttgcgc gtacgcgttc catgtggtca ttcaatccag aaactaacat tcttctcaac
      841 gtgccactcc atggcactat tctgaccaga ccgcttctag aaagtgaact cgtaatcgga
      901 gctgtgatcc ttcgtggaca tcttcgtatt gctggacacc atctaggacg ctgtgacatc
      961 aaggacctgc ctaaagaaat cactgttgct acatcacgaa cgctttctta ttacaaattg
     1021 ggagcttcgc agcgtgtagc aggtgactca ggttttgctg catacagtcg ctacaggatt
     1081 ggcaactata aattaaacac agaccattcc agtagcagtg acaatattgc tttgcttgta
     1141 cagctcgtgc cgagaggttc ttggagccat ccgcagtttg aaaaataaac aggccccttt
     1201 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tcctcccaca
     1261 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     1321 ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     1381 tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct tgaaattttt
     1441 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     1501 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     1561 tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct acatataagg
     1621 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa
     1681 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag
     1741 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt
     1801 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac
     1861 tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg
     1921 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag
     1981 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt
     2041 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta
     2101 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca
     2161 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg
     2221 acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt tctacaggat
     2281 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg
     2341 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
     2401 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     2461 aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa
     2521 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     2581 aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac
     2641 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     2701 aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt
     2761 tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt
     2821 ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt
     2881 aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc
     2941 acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca
     3001 tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag
     3061 cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata
     3121 tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt
     3181 cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg
     3241 tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata
     3301 gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca
     3361 atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag
     3421 agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga
     3481 acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact gcaatcaact
     3541 cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga tcttcttgag
     3601 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3661 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg gcttcagcag
     3721 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3781 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3841 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3901 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3961 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4021 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4081 aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagca
     4141 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaga
     4201 aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga ggttttcacc
     4261 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     4321 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     4381 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactta
     4441 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4501 tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat ctggccccag
     4561 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4621 gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct ccatccagtc
     4681 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4741 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4801 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4861 tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt tatcactcat
     4921 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat gcttttctgt
     4981 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5041 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5101 cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag
     5161 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5221 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5281 gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     5341 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     5401 aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca ggtatgattt
     5461 aaatggtcag ttaga
//
