LOCUS       sequence-297939-        6549 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 6549)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6549)
  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-m4046225
FEATURES             Location/Qualifiers
     source          1..6549
                     /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"
     primer_bind     complement(2304..2322)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2673..3476
                     /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      3559..4541
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4619..4648
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4670..5257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5269..5296)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5320..5511)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5512..6303,6304..6372))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESXRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6135..6154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6373..6475)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6476..6507)
                     /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 taaaatcgag gaaggtaagc tagtcatttg gataaatgga
     1201 gataagggtt acaacggttt ggcagaagta ggcaagaaat ttgaaaagga cacaggtatc
     1261 aaggtgacgg ttgaacatcc agacaagtta gaagaaaaat tccctcaagt tgctgctact
     1321 ggtgatggtc ccgacattat tttctgggcc cacgatcgtt ttggtggata tgctcagtca
     1381 ggtttgctgg ctgagatcac cccagacaag gccttccaag ataaattgta cccattcact
     1441 tgggatgctg tcagatacaa cggtaagctt atagcatatc caattgctgt tgaagcctta
     1501 tcgttgatct acaataagga cttgttacct aacccaccaa agacatggga agaaattcca
     1561 gctttggata aagagctaaa ggccaaggga aaaagcgctt tgatgtttaa cttacaagaa
     1621 ccttatttca cctggccatt aatcgctgct gacggtggtt acgcatttaa atacgaaaac
     1681 ggtaagtatg acattaagga tgtcggcgtt gataatgctg gtgcaaaggc tggtttgacc
     1741 ttcttggttg atttaataaa gaataaacac atgaacgctg acactgacta ctctatcgcc
     1801 gaagccgctt tcaacaaggg tgaaactgct atgaccataa atggtccatg ggcctggagt
     1861 aacattgata cttctaaagt aaactatggt gtcactgttt tgcctacatt caaaggccaa
     1921 ccatccaagc cctttgtcgg tgttctatcc gctggcatca atgctgcttc tccaaacaag
     1981 gagctagcta aagaattttt ggaaaattac ttgttaactg acgaaggttt ggaggccgtt
     2041 aacaaggaca agccattggg tgctgtcgct ttgaaatctt acgaagaaga attggctaag
     2101 gatcccagaa ttgccgctac catggagaac gcccaaaagg gtgaaattat gccaaacatc
     2161 ccacaaatgt cagccttctg gtacgcagtt aggactgccg tgattaatgc tgcttccggt
     2221 agacaaaccg tcgacgaagc attaaaagat gctcaaacgt aaacaggccc cttttccttt
     2281 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     2341 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     2401 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     2461 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     2521 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     2581 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     2641 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     2701 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     2761 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     2821 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     2881 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     2941 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     3001 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     3061 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     3121 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     3181 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     3241 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     3301 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     3361 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     3421 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     3481 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     3541 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     3601 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     3661 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     3721 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     3781 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     3841 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     3901 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     3961 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     4021 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     4081 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     4141 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     4201 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     4261 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     4321 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     4381 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     4441 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     4501 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     4561 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     4621 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     4681 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4741 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     4801 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4861 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4921 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4981 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5041 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5101 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     5161 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     5221 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     5281 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     5341 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     5401 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5461 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     5521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5581 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     5641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5701 cggragggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     5761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5941 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     6001 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     6061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6181 aaaacgttct tcggggcgwa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6361 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     6421 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     6481 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     6541 tcagttaga
//
