LOCUS       sequence-298353-        5470 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 5470)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5470)
  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-m4046457
FEATURES             Location/Qualifiers
     source          1..5470
                     /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..1180
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1225..1243)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1594..2397
                     /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      2480..3462
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3540..3569
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3591..4178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4190..4217)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4241..4432)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4433..5224,5225..5293))
                     /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     5056..5075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5294..5396)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5397..5428)
                     /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 atggctgatt ccaatggcac catcacagta gaagagctga aaaagctcct
      481 tgaacaatgg aacttggtta taggtttttt attcttgact tggatttgcc tattgcagtt
      541 cgcctatgca aacagaaacc gttttttgta cattatcaag ttaattttct tgtggctgtt
      601 atggcctgtc actcttgctt gttttgtttt ggctgccgtc taccggataa attggatcac
      661 cggtggtatc gctattgcaa tggcttgttt ggttggattg atgtggttga gttatttcat
      721 tgcctcattc agattattcg ctaggactag atcgatgtgg tcctttaatc cagaaacgaa
      781 catcttgttg aacgtgccac tccatggtac tattttgacc agaccgctat tagaatctga
      841 gttggttatc ggtgctgtca ttctaagagg gcacttacgt atcgcgggtc accatttggg
      901 aagatgtgac attaaggatc ttccaaaaga aatcacagtc gctacctctc gaactctatc
      961 ttactacaag ttgggtgcct cacaaagggt tgctggtgac agcggtttcg ctgcctattc
     1021 tcgctaccgt atcggcaact acaaattaaa taccgatcac tcctctagtt ctgacaatat
     1081 tgccttattg gtgcaactcg tgccgagagg ttctgattat aaagatcatg atggtgatta
     1141 taaagatcat gatattgatt ataaagatga tgatgataaa taaacaggcc ccttttcctt
     1201 tgtcgatatc atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc
     1261 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta
     1321 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     1381 aaagcggccg ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat
     1441 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     1501 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     1561 agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     1621 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     1681 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     1741 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc
     1801 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     1861 gaggacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     1921 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt
     1981 gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg
     2041 ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt
     2101 gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt
     2161 ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag
     2221 ggagatgcat tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac
     2281 attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     2341 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     2401 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     2461 tatcagttat tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     2521 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     2581 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     2641 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     2701 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     2761 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     2821 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     2881 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     2941 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     3001 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     3061 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     3121 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     3181 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     3241 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     3301 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
     3361 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc
     3421 ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc
     3481 ccgagcggcc gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg
     3541 gctcaccttc acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct
     3601 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3661 tgtttgccgg atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc
     3721 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3781 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3841 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3901 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3961 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4021 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4081 taaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat
     4141 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc
     4201 ccacccgaag gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat
     4261 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     4321 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     4381 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat
     4441 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4501 gactccccgt cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg
     4561 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4621 ccgggagggc cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta
     4681 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4741 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4801 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4861 ccttcggtcc tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta
     4921 tggcagcact gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg
     4981 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5041 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5101 gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5161 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5221 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5281 gttgaatact cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     5341 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc
     5401 aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg
     5461 gtcagttaga
//
