LOCUS       sequence-291515-        5428 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 5428)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5428)
  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-m4046292
FEATURES             Location/Qualifiers
     source          1..5428
                     /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..1138
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1183..1201)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1552..2355
                     /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      2438..3420
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3498..3527
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3549..4136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4148..4175)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4199..4390)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4391..5182,5183..5251))
                     /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     5014..5033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5252..5354)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5355..5386)
                     /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 ttcttggagc catccgcagt ttgaaaaata
     1141 aacaggcccc ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg
     1201 ccctcctccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     1261 ccctatttat tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt
     1321 tttctttttt ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt
     1381 ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac
     1441 agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta
     1501 ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa
     1561 gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat
     1621 atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa
     1681 ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat
     1741 atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag
     1801 tacaattttt tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg
     1861 cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt
     1921 gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa
     1981 cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa
     2041 tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt
     2101 gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt
     2161 gtgggtttag atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg
     2221 gtttctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat
     2281 gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc
     2341 ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt
     2401 agagcttcaa tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt
     2461 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     2521 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
     2581 atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     2641 tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat
     2701 acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct
     2761 tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca
     2821 ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa
     2881 aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt
     2941 caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa
     3001 cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta
     3061 ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac
     3121 tctatgaata gttcttacta caattttttt gtctaaagag taatactaga gataaacata
     3181 aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt
     3241 atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga
     3301 agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa
     3361 agtgcgtcat cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag
     3421 ctagagaata ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg
     3481 actgcaatca actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa
     3541 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3601 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa
     3661 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3721 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3781 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3841 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3901 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3961 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4021 cgagggagct tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4081 tctgacttga gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4141 ccagcaacgc agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc
     4201 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     4261 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     4321 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     4381 gtttggtcac ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4441 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac
     4501 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4561 cagcaataaa ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg
     4621 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4681 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4741 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4801 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag
     4861 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga
     4921 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4981 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5041 taaaagtgct catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc
     5101 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5161 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5221 taagggcgac acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg
     5281 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     5341 ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg
     5401 tcaggtatga tttaaatggt cagttaga
//
