LOCUS       sequence-297979-        5429 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 5429)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5429)
  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-m4046292
FEATURES             Location/Qualifiers
     source          1..5429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..418
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1098..1115
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1116..1139
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1184..1202)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1553..2356
                     /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      2439..3421
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3499..3528
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3550..4137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4149..4176)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4200..4391)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4392..5183,5184..5252))
                     /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     5015..5034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5253..5355)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5356..5387)
                     /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 ttttttttng atttttttct ctttcgatga cctcccattg
      301 atatttaagt taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct
      361 attacaactt tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt
      421 ttaattacaa aatggcagat tccaacggta ctattaccgt tgaagagctt aaaaagctcc
      481 ttgaacaatg gaacctagta ataggtttcc tattccttac atggatttgt cttctacaat
      541 ttgcctatgc caacaggaat aggtttttgt atataattaa gttaattttc ctctggctgt
      601 tatggccagt aactttagct tgttttgtgc ttgctgctgt ttacagaata aattggatca
      661 ccggtggaat tgctatcgca atggcttgtc ttgtaggctt gatgtggctc agctacttca
      721 ttgcttcttt cagactgttt gcgcgtacgc gttccatgtg gtcattcaat ccagaaacta
      781 acattcttct caacgtgcca ctccatggca ctattctgac cagaccgctt ctagaaagtg
      841 aactcgtaat cggagctgtg atccttcgtg gacatcttcg tattgctgga caccatctag
      901 gacgctgtga catcaaggac ctgcctaaag aaatcactgt tgctacatca cgaacgcttt
      961 cttattacaa attgggagct tcgcagcgtg tagcaggtga ctcaggtttt gctgcataca
     1021 gtcgctacag gattggcaac tataaattaa acacagacca ttccagtagc agtgacaata
     1081 ttgctttgct tgtacagctc gtgccgagag gttcttggag ccatccgcag tttgaaaaat
     1141 aaacaggccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac
     1201 gccctcctcc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     1261 tccctattta ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat
     1321 ttttcttttt tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt
     1381 tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca
     1441 cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt
     1501 attcttaacc caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa
     1561 agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa
     1621 tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga
     1681 attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga
     1741 tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa
     1801 gtacaatttt ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt
     1861 gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg
     1921 tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga
     1981 acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga
     2041 atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat
     2101 tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg
     2161 tgtgggttta gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt
     2221 ggtttctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga
     2281 tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg
     2341 cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat
     2401 tagagcttca atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt
     2461 ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     2521 tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt
     2581 catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     2641 ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta
     2701 tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc
     2761 ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc
     2821 actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa
     2881 aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt
     2941 tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga
     3001 acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct
     3061 attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca
     3121 ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat
     3181 aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt
     3241 tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg
     3301 aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga
     3361 aagtgcgtca tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta
     3421 gctagagaat aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg
     3481 gactgcaatc aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa
     3541 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3601 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta
     3661 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3721 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3781 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3841 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3901 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3961 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4021 acgagggagc ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4081 ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4141 gccagcaacg cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat
     4201 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     4261 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     4321 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     4381 tgtttggtca cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4441 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta
     4501 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4561 tcagcaataa accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc
     4621 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4681 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4741 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4801 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca
     4861 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg
     4921 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4981 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5041 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg
     5101 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5161 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5221 ataagggcga cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc
     5281 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     5341 cttacgtgcc gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag
     5401 gtcaggtatg atttaaatgg tcagttaga
//
