LOCUS       sequence-298323-        5469 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 5469)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5469)
  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-m4046216
FEATURES             Location/Qualifiers
     source          1..5469
                     /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..1179
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1224..1242)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1593..2396
                     /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      2479..3461
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3539..3568
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3590..4177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4189..4216)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4240..4431)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4432..5223,5224..5292))
                     /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     5055..5074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5293..5395)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5396..5427)
                     /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 tcaccatcat caccatcatt aaacaggccc cttttccttt
     1201 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     1261 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     1321 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     1381 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     1441 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     1501 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     1561 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     1621 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     1681 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     1741 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     1801 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     1861 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     1921 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     1981 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     2041 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     2101 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     2161 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     2221 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     2281 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     2341 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     2401 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     2461 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     2521 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2581 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     2641 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     2701 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     2761 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     2821 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     2881 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     2941 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     3001 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     3061 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     3121 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     3181 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     3241 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     3301 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     3361 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     3421 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     3481 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     3541 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     3601 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3661 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     3721 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3781 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3841 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3901 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3961 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4021 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     4081 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     4141 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     4201 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     4261 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     4321 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     4381 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     4441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4501 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     4561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4621 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     4681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4861 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     4921 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     4981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5101 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5281 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     5341 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     5401 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     5461 tcagttaga
//
