LOCUS       sequence-297774-        6469 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 M (membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6469)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6469)
  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-m4046310
FEATURES             Location/Qualifiers
     source          1..6469
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        4..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1097..1114
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2257..2275)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2626..3429
                     /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"
     terminator      4539..4568
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4590..5177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5189..5216)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5240..5431)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5432..6223,6224..6292))
                     /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     6055..6074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6293..6395)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6396..6427)
                     /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 atggcagatt ccaacggtac tattaccgtt gaagagctta aaaagctcct
      481 tgaacaatgg aacctagtaa taggtttcct attccttaca tggatttgtc ttctacaatt
      541 tgcctatgcc aacaggaata ggtttttgta tataattaag ttaattttcc tctggctgtt
      601 atggccagta actttagctt gttttgtgct tgctgctgtt tacagaataa attggatcac
      661 cggtggaatt gctatcgcaa tggcttgtct tgtaggcttg atgtggctca gctacttcat
      721 tgcttctttc agactgtttg cgcgtacgcg ttccatgtgg tcattcaatc cagaaactaa
      781 cattcttctc aacgtgccac tccatggcac tattctgacc agaccgcttc tagaaagtga
      841 actcgtaatc ggagctgtga tccttcgtgg acatcttcgt attgctggac accatctagg
      901 acgctgtgac atcaaggacc tgcctaaaga aatcactgtt gctacatcac gaacgctttc
      961 ttattacaaa ttgggagctt cgcagcgtgt agcaggtgac tcaggttttg ctgcatacag
     1021 tcgctacagg attggcaact ataaattaaa cacagaccat tccagtagca gtgacaatat
     1081 tgctttgctt gtacagctcg tgccgagagg ttctaaaatc gaggaaggta agctagtcat
     1141 ttggataaat ggagataagg gttacaacgg tttggcagaa gtaggcaaga aatttgaaaa
     1201 ggacacaggt atcaaggtga cggttgaaca tccagacaag ttagaagaaa aattccctca
     1261 agttgctgct actggtgatg gtcccgacat tattttctgg gcccacgatc gttttggtgg
     1321 atatgctcag tcaggtttgc tggctgagat caccccagac aaggccttcc aagataaatt
     1381 gtacccattc acttgggatg ctgtcagata caacggtaag cttatagcat atccaattgc
     1441 tgttgaagcc ttatcgttga tctacaataa ggacttgtta cctaacccac caaagacatg
     1501 ggaagaaatt ccagctttgg ataaagagct aaaggccaag ggaaaaagcg ctttgatgtt
     1561 taacttacaa gaaccttatt tcacctggcc attaatcgct gctgacggtg gttacgcatt
     1621 taaatacgaa aacggtaagt atgacattaa ggatgtcggc gttgataatg ctggtgcaaa
     1681 ggctggtttg accttcttgg ttgatttaat aaagaataaa cacatgaacg ctgacactga
     1741 ctactctatc gccgaagccg ctttcaacaa gggtgaaact gctatgacca taaatggtcc
     1801 atgggcctgg agtaacattg atacttctaa agtaaactat ggtgtcactg ttttgcctac
     1861 attcaaaggc caaccatcca agccctttgt cggtgttcta tccgctggca tcaatgctgc
     1921 ttctccaaac aaggagctag ctaaagaatt tttggaaaat tacttgttaa ctgacgaagg
     1981 tttggaggcc gttaacaagg acaagccatt gggtgctgtc gctttgaaat cttacgaaga
     2041 agaattggct aaggatccca gaattgccgc taccatggag aacgcccaaa agggtgaaat
     2101 tatgccaaac atcccacaaa tgtcagcctt ctggtacgca gttaggactg ccgtgattaa
     2161 tgctgcttcc ggtagacaaa ccgtcgacga agcattaaaa gatgctcaaa cgtaaacagg
     2221 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     2281 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     2341 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     2401 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     2461 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     2521 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     2581 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     2641 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     2701 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     2761 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     2821 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     2881 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     2941 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     3001 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     3061 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     3121 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     3181 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     3241 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     3301 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     3361 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     3421 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     3481 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     3541 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     3601 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     3661 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3721 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacgcgc
     3781 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
     3841 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
     3901 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
     3961 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
     4021 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
     4081 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
     4141 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
     4201 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
     4261 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
     4321 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg
     4381 gtgcgttttt ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc
     4441 tgaagttcct atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa
     4501 gcttgactcc agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc
     4561 tttcttcggt agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4621 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4681 ccaactcttt ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4741 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4801 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4861 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4921 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4981 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5041 gggtcggaac aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata
     5101 gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg
     5161 ggcggagcct atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt
     5221 gattacattt gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     5281 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     5341 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     5401 tgttaagggc ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta
     5461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5521 ctacgatgcg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5581 gctcaccggc tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa
     5641 gtgatcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5881 ccagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5941 ttactgtcat gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat
     6001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta
     6121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt
     6301 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     6361 tatggtgaaa gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct
     6421 tttgactttc tgctatggag gtcaggtatg atttaaatgg tcagttaga
//
