LOCUS       sequence-291475-        6502 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 6502)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6502)
  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..6502
                     /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"
     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"
     rep_origin      3512..4494
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4572..4601
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4623..5210
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5222..5249)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5273..5464)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5465..6256,6257..6325))
                     /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     6088..6107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6326..6428)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6429..6460)
                     /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 ctatacttct
     3781 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     3841 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     3901 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     3961 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     4021 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     4081 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     4141 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     4201 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     4261 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     4321 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     4381 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     4441 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     4501 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     4561 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     4621 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4681 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     4741 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4801 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4861 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4921 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4981 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5041 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5101 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5161 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5221 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     5281 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     5341 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     5401 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     5461 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     5581 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5641 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     5701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5881 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     5941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     6001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6121 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     6181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6301 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     6361 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     6421 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     6481 atgatttaaa tggtcagtta ga
//
