LOCUS       sequence-291510-        5422 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 5422)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5422)
  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-m4046428
FEATURES             Location/Qualifiers
     source          1..5422
                     /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             1097..1114
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1115..1132
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1177..1195)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1546..2349
                     /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      2432..3414
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3492..3521
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3543..4130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4142..4169)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4193..4384)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4385..5176,5177..5245))
                     /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     5008..5027
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5246..5348)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5349..5380)
                     /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 ttctcaccat catcaccatc attaaacagg
     1141 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     1201 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     1261 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     1321 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     1381 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     1441 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     1501 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     1561 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     1621 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     1681 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     1741 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     1801 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     1861 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     1921 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     1981 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     2041 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     2101 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     2161 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     2221 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     2281 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     2341 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     2401 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     2461 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     2521 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     2581 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2641 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     2701 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     2761 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     2821 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     2881 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     2941 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     3001 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     3061 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     3121 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     3181 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     3241 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     3301 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     3361 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     3421 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     3481 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     3541 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3601 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     3661 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3721 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3781 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3841 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3901 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3961 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4021 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4081 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4141 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     4201 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     4261 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     4321 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     4381 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4441 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     4501 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4561 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     4621 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4681 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4741 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4801 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     4861 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     4921 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4981 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5041 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     5101 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5161 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5221 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     5281 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     5341 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     5401 atgatttaaa tggtcagtta ga
//
