LOCUS       sequence-307321-        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-m4046253
FEATURES             Location/Qualifiers
     source          1..5428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..414
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1094..1111
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1112..1135
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1180..1198)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1549..2352
                     /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      2435..3417
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3495..3524
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3546..4133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4145..4172)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4196..4387)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4388..5179,5180..5248))
                     /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     5011..5030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5249..5351)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5352..5383)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gccgccccac acaccatagc ttcaaaatgt ttctactcct tttttactct tccagatttt
       61 ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaatttc
      121 ccctctttct tcctctaggg tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa
      181 agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt
      241 ctttttcttg aaaatttttt tttttgattt ttttctcttt cgatgacctc ccattgatat
      301 ttaagttaat aaacggtctt caatttctca agtttcagtt tcatttttct tgttctatta
      361 caactttttt tacttcttgc tcattagaaa gaaagcatag caatctaatc taagttttaa
      421 ttacaaaatg gctgattcca atggcaccat cacagtagaa gagctgaaaa agctccttga
      481 acaatggaac ttggttatag gttttttatt cttgacttgg atttgcctat tgcagttcgc
      541 ctatgcaaac agaaaccgtt ttttgtacat tatcaagtta attttcttgt ggctgttatg
      601 gcctgtcact cttgcttgtt ttgttttggc tgccgtctac cggataaatt ggatcaccgg
      661 tggtatcgct attgcaatgg cttgtttggt tggattgatg tggttgagtt atttcattgc
      721 ctcattcaga ttattcgcta ggactagatc gatgtggtcc tttaatccag aaacgaacat
      781 cttgttgaac gtgccactcc atggtactat tttgaccaga ccgctattag aatctgagtt
      841 ggttatcggt gctgtcattc taagagggca cttacgtatc gcgggtcacc atttgggaag
      901 atgtgacatt aaggatcttc caaaagaaat cacagtcgct acctctcgaa ctctatctta
      961 ctacaagttg ggtgcctcac aaagggttgc tggtgacagc ggtttcgctg cctattctcg
     1021 ctaccgtatc ggcaactaca aattaaatac cgatcactcc tctagttctg acaatattgc
     1081 cttattggtg caactcgtgc cgagaggttc ttggagccat ccgcagtttg aaaaataaac
     1141 aggccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc
     1201 tcctcccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     1261 tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     1321 cttttttttc tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct
     1381 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     1441 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     1501 ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct
     1561 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     1621 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     1681 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     1741 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     1801 aattttttac tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag
     1861 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     1921 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     1981 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     2041 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     2101 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     2161 ggtttagatg acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt
     2221 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     2281 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     2341 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     2401 gcttcaattt aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt
     2461 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     2521 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     2581 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2641 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     2701 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     2761 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     2821 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     2881 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     2941 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     3001 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     3061 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     3121 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     3181 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     3241 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     3301 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     3361 gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     3421 gagaatagga acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact
     3481 gcaatcaact cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga
     3541 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3601 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg
     3661 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3721 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3781 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3841 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3901 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3961 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4021 gggagcttcc aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4081 gacttgagca tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4141 gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga
     4201 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     4261 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4321 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4381 tggtcactta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4441 catccatagt tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat
     4501 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4561 caataaacca gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct
     4621 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4681 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4741 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4801 aaaaagcggt tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt
     4861 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat
     4921 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4981 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5041 aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt
     5101 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5161 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5221 gggcgacacg gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata
     5281 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     5341 cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca
     5401 ggtatgattt aaatggtcag ttagagcg
//
