LOCUS       sequence-297606-        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"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1141..1159)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1510..2313
                     /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      2396..3378
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3456..3485
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3507..4094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4106..4133)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4157..4348)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4349..5140,5141..5209))
                     /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     4972..4991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5210..5312)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5313..5344)
                     /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 ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggctgatt ccaatggcac catcacagta gaagagctga aaaagctcct
      481 tgaacaatgg aacttggtta taggtttttt attcttgact tggatttgcc tattgcagtt
      541 cgcctatgca aacagaaacc gttttttgta cattatcaag ttaattttct tgtggctgtt
      601 atggcctgtc actcttgctt gttttgtttt ggctgccgtc taccggataa attggatcac
      661 cggtggtatc gctattgcaa tggcttgttt ggttggattg atgtggttga gttatttcat
      721 tgcctcattc agattattcg ctaggactag atcgatgtgg tcctttaatc cagaaacgaa
      781 catcttgttg aacgtgccac tccatggtac tattttgacc agaccgctat tagaatctga
      841 gttggttatc ggtgctgtca ttctaagagg gcacttacgt atcgcgggtc accatttggg
      901 aagatgtgac attaaggatc ttccaaaaga aatcacagtc gctacctctc gaactctatc
      961 ttactacaag ttgggtgcct cacaaagggt tgctggtgac agcggtttcg ctgcctattc
     1021 tcgctaccgt atcggcaact acaaattaaa taccgatcac tcctctagtt ctgacaatat
     1081 tgccttattg gtgcaataaa caggcccctt ttcctttgtc gatatcatgt aattagttat
     1141 gtcacgctta cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag
     1201 acaacctgaa gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg
     1261 ttatttatat ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg
     1321 cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa
     1381 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
     1441 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga
     1501 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt
     1561 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat
     1621 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta
     1681 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta
     1741 aaggcattat ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt
     1801 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac
     1861 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca
     1921 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc
     1981 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat
     2041 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg
     2101 ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat
     2161 agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta
     2221 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     2281 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     2341 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga
     2401 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     2461 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     2521 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     2581 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     2641 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     2701 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     2761 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     2821 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     2881 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     2941 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     3001 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     3061 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     3121 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     3181 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     3241 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     3301 cgtttttggt tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga
     3361 agttcctata ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct
     3421 tgactccagc gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt
     3481 cttcggtaga aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3541 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3601 actctttttc cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3961 tcggaacagg agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc
     4021 ctgtcgggtt tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc
     4081 ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat
     4141 tacatttggg ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     4201 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4261 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4321 taagggcggt tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct
     4381 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4441 cgatgcggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4501 caccggctcc agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg
     4561 atcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4621 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4681 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4741 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca
     4801 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4861 ctgtcatgcc atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4921 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4981 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac
     5041 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5101 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5161 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct
     5221 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     5281 ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt
     5341 gactttctgc tatggaggtc aggtatgatt taaatggtca gttaga
//
