LOCUS       sequence-298356-        5517 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 5517)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5517)
  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-m4046277
FEATURES             Location/Qualifiers
     source          1..5517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1144..1161
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1162..1227
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1272..1290)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1641..2444
                     /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      2527..3509
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3587..3616
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3638..4225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4237..4264)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4288..4479)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4480..5271,5272..5340))
                     /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     5103..5122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5341..5443)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5444..5475)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gctgattcca atggcaccat cacagtagaa gagctgaaaa agctccttga acaatggaac
      541 ttggttatag gttttttatt cttgacttgg atttgcctat tgcagttcgc ctatgcaaac
      601 agaaaccgtt ttttgtacat tatcaagtta attttcttgt ggctgttatg gcctgtcact
      661 cttgcttgtt ttgttttggc tgccgtctac cggataaatt ggatcaccgg tggtatcgct
      721 attgcaatgg cttgtttggt tggattgatg tggttgagtt atttcattgc ctcattcaga
      781 ttattcgcta ggactagatc gatgtggtcc tttaatccag aaacgaacat cttgttgaac
      841 gtgccactcc atggtactat tttgaccaga ccgctattag aatctgagtt ggttatcggt
      901 gctgtcattc taagagggca cttacgtatc gcgggtcacc atttgggaag atgtgacatt
      961 aaggatcttc caaaagaaat cacagtcgct acctctcgaa ctctatctta ctacaagttg
     1021 ggtgcctcac aaagggttgc tggtgacagc ggtttcgctg cctattctcg ctaccgtatc
     1081 ggcaactaca aattaaatac cgatcactcc tctagttctg acaatattgc cttattggtg
     1141 caactcgtgc cgagaggttc tgattataaa gatcatgatg gtgattataa agatcatgat
     1201 attgattata aagatgatga tgataaataa acaggcccct tttcctttgt cgatatcatg
     1261 taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct aaccgaaaag
     1321 gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata gttatgttag
     1381 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa gcggccgccg
     1441 aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg
     1501 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     1561 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt
     1621 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
     1681 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
     1741 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
     1801 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
     1861 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat
     1921 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
     1981 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
     2041 agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
     2101 catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga
     2161 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
     2221 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg
     2281 gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg
     2341 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     2401 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     2461 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     2521 ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     2581 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     2641 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     2701 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     2761 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     2821 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     2881 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     2941 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     3001 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     3061 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     3121 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
     3181 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
     3241 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
     3301 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
     3361 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
     3421 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa
     3481 aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg agcggccgcg
     3541 tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg
     3601 ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg
     3661 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3721 aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac
     3781 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3841 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3901 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3961 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4021 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4081 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa acgcctggta
     4141 tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc
     4201 gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg
     4261 agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac cgaaacgcgc
     4321 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     4381 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     4441 gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct taatcagtga
     4501 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4561 gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4621 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga
     4681 gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4741 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4801 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4861 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4921 gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4981 taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac
     5041 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5101 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5161 ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5221 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5281 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5341 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     5401 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt
     5461 cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc agttaga
//
