LOCUS       sequence-297959-        5706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 2'-O-ribose 
            methyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5706)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5706)
  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-m4046556
FEATURES             Location/Qualifiers
     source          1..5706
                     /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             1375..1392
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1393..1416
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1461..1479)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1830..2633
                     /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      2716..3698
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3776..3805
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3827..4414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4426..4453)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4477..4668)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4669..5460,5461..5529))
                     /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     5292..5311
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5530..5632)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5633..5664)
                     /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 tcctctcaag cctggcagcc tggtgtggct atgcccaatt tgtataagat gcaaagaatg
      541 cttctggaga aatgcgattt gcaaaactac ggagacagcg ctacattacc aaagggtatt
      601 atgatgaacg tagcaaagta cacccaattg tgtcaatact taaatacgtt gactttggct
      661 gttccgtata acatgcgtgt catacacttt ggggccggtt cagacaaagg cgtcgctcca
      721 ggtaccgcag ttctaaggca atggctccca actggtactc tattggttga ttctgatctg
      781 aacgacttcg tcagtgatgc tgactccact ttaatcggtg attgtgctac cgttcatacg
      841 gcgaacaagt gggacttgat tatttcggat atgtacgacc caaagaccaa aaatgttaca
      901 aaggaaaatg attctaaaga aggtttcttt acttatatct gtggtttcat ccagcaaaag
      961 cttgccttgg gcggttctgt ggctattaag atcactgaac actcctggaa cgccgactta
     1021 tacaaattga tgggtcattt cgcttggtgg accgcgtttg ttaccaacgt taacgctagt
     1081 tcttcggaag cctttctcat cggttgtaat tatttaggaa agccacgcga acaaattgat
     1141 ggttacgtca tgcacgctaa ctacattttc tggagaaata ctaaccctat acaattgtca
     1201 agctatagtt tattcgacat gtcaaagttt ccattgaagt tgagaggtac agctgtcatg
     1261 tccttgaagg aagggcagat caacgatatg atcttatctt tgctgtctaa aggtagactc
     1321 attattagag aaaataatcg ggttgttatc tcttctgatg tgctagtcaa caacctcgtg
     1381 ccgagaggtt cttggagcca tccgcagttt gaaaaataaa caggcccctt ttcctttgtc
     1441 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     1501 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     1561 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     1621 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     1681 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     1741 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     1801 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     1861 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     1921 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     1981 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     2041 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     2101 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     2161 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     2221 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     2281 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     2341 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     2401 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     2461 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     2521 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     2581 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     2641 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     2701 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     2761 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2821 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     2881 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     2941 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     3001 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     3061 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     3121 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     3181 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     3241 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     3301 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3361 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     3421 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     3481 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     3541 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     3601 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     3661 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     3721 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     3781 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     3841 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3901 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     3961 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4021 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4081 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4141 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4201 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4261 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     4321 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     4381 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     4441 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     4501 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4561 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4621 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     4681 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4741 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     4801 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4861 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     4921 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4981 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5041 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5101 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5161 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     5221 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5281 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5341 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5401 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5461 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5521 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     5581 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     5641 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     5701 gttaga
//
