LOCUS       sequence-300485-        5386 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 5386)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5386)
  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-m4046617
FEATURES             Location/Qualifiers
     source          1..5386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(366..1169)
                     /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"
     primer_bind     1520..1538
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(2262..2675)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      2721..2752
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2753..2855
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(2856..2924,2925..3716)
                     /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     complement(3074..3093)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3717..3908
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      3932..3959
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3971..4558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4580..4609)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(4687..5386,1..283)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 ttgtagaaca aaaaagaagt atagattctt tgttggtaaa atagcgctct cgcgttgcat
       61 ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg
      121 catttttgtt ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt
      181 tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg
      241 cgttgcattt ttgttctaca aaatgaagca cagatgcttc gttactcggg taataactga
      301 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
      361 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
      421 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
      481 atgtcagatc ctgtagaaac cacatcatcc acggttctat actgttgacc caatgcatct
      541 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
      601 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
      661 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
      721 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
      781 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
      841 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtcc
      901 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
      961 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     1021 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     1081 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     1141 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca cgtttttgtt
     1201 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     1261 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     1321 gaatcaaaaa aatttcaagg aaaccgtgta ctgcagtgca ctagtggttc ggcggccgct
     1381 ttgtacagaa aaaaaagaaa aatttgaaat ataaataacg ttcttaatac taacataact
     1441 attaaaaaaa ataaataggg acctagactt caggttgtct aactccttcc ttttcggtta
     1501 gagcggatgt gggaggaggg cgtgaatgta agcgtgacat aactaattac atgatatcga
     1561 caaaggaaaa ggggcctgtt tattgcacca ataaggcaat attgtcagaa ctagaggagt
     1621 gatcggtatt taatttgtag ttgccgatac ggtagcgaga ataggcagcg aaaccgctgt
     1681 caccagcaac cctttgtgag gcacccaact tgtagtaaga tagagttcga gaggtagcga
     1741 ctgtgatttc ttttggaaga tccttaatgt cacatcttcc caaatggtga cccgcgatac
     1801 gtaagtgccc tcttagaatg acagcaccga taaccaactc agattctaat agcggtctgg
     1861 tcaaaatagt accatggagt ggcacgttca acaagatgtt cgtttctgga ttaaaggacc
     1921 acatcgatct agtcctagcg aataatctga atgaggcaat gaaataactc aaccacatca
     1981 atccaaccaa acaagccatt gcaatagcga taccaccggt gatccaattt atccggtaga
     2041 cggcagccaa aacaaaacaa gcaagagtga caggccataa cagccacaag aaaattaact
     2101 tgataatgta caaaaaacgg tttctgtttg cataggcgaa ctgcaatagg caaatccaag
     2161 tcaagaataa aaaacctata accaagttcc attgttcaag gagctttttc agctcttcta
     2221 ctgtgatggt gccattggaa tcagccattt tgtaattaaa acttagatta gattgctatg
     2281 ctttctttct aatgagcaag aagtaaaaaa agttgtaata gaacaagaaa aatgaaactg
     2341 aaacttgaga aattgaagac cgtttattaa cttaaatatc aatgggaggt catcgaaaga
     2401 gaaaaaaatc aaaaaaaaaa attttcaaga aaaagaaacg tgataaaaat ttttattgcc
     2461 tttttcgacg aagaaaaaga aacgaggcgg tctctttttt cttttccaaa cctttagtac
     2521 gggtaattaa cgacacccta gaggaagaaa gaggggaaat ttagtatgct gtgcttgggt
     2581 gttttgaagt ggtacggcga tgcgcggagt ccgagaaaat ctggaagagt aaaaaaggag
     2641 tagaaacatt ttgaagctat ggtgtgtggg gcggccgctc taactgacca tttaaatcat
     2701 acctgacctc catagcagaa agtcaaaagc ctccgaccgg aggcttttga cttgatcggc
     2761 acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg
     2821 agttatcgag attttcagga gctaaggaag ctaaaatgag tattcaacat ttccgtgtcg
     2881 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2941 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3001 tcaacagcgg taagatcctt gagagtttac gccccgaaga acgttttcca atgatgagca
     3061 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3121 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3181 agcatctcac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3241 ataacactgc ggccaactta cttctggcaa cgatcggagg accgaaggag ctaaccgctt
     3301 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3361 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3421 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3481 tggaggcgga taaagttgca ggatcacttc tgcgctcggc cctcccggct ggctggttta
     3541 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3601 cagatggtaa gccctcccgc atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3661 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaagtga
     3721 ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc
     3781 ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc
     3841 acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa
     3901 acctctgatg agggcccaaa tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt
     3961 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa
     4021 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttacc cctggaagct
     4081 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4141 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4201 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4261 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4321 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4381 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4441 agccagttac ctcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4501 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4561 agatcctttg attttctacc gaagaaaggc ccacccgtga aggtgagcca gtgagttgat
     4621 tgcagtccag ttacgctgga gtcaagcttg taacacgcgg ccgctcggga agttcctatt
     4681 ctctagctag aaagtatagg aacttcagag cgcttttgaa aaccaaaagc gctctgatga
     4741 cgcactttca aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa tattgcgaat
     4801 accgcttcca caaacattgc tcaaaagtat ctctttgcta tatatctctg tgctatatcc
     4861 ctatataacc tacccatcca cctttcgctc cttgaacttg catctaaact cgacctctac
     4921 attttttatg tttatctcta gtattactct ttagacaaaa aaattgtagt aagaactatt
     4981 catagagtga atcgaaaaca atacgaaaat gtaaacattt cctatacgta gtatatagag
     5041 acaaaataga agaaaccgtt cataattttc tgaccaatga agaatcatca acgctatcac
     5101 tttctgttca caaagtatgc gcaatccaca tcggtataga atataatcgg ggatgccttt
     5161 atcttgaaaa aatgcacccg cagcttcgct agtaatcagt aaacgcggga agtggagtca
     5221 ggcttttttt atggaagaga aaatagacac caaagtagcc ttcttctaac cttaacggac
     5281 ctacagtgca aaaagttatc aagagactgc attatagagc gcacaaagga gaaaaaaagt
     5341 aatctaagat gctttgttag aaaaatagcg ctctcgggat gcattt
//
