LOCUS       sequence-287668-        5742 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 5742)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5742)
  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-m4046364
FEATURES             Location/Qualifiers
     source          1..5742
                     /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             1369..1386
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1387..1452
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1497..1515)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1866..2669
                     /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      2752..3734
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3812..3841
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3863..4450
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4462..4489)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4513..4704)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4705..5496,5497..5565))
                     /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     5328..5347
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5566..5668)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5669..5700)
                     /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 agtcaagcgt ggcaaccggg tgttgctatg cctaatcttt acaaaatgca aagaatgcta
      541 ttagaaaagt gtgaccttca aaattatggt gatagtgcaa cattacctaa aggcataatg
      601 atgaatgtcg caaaatatac tcaactgtgt caatatttaa acacattaac attagctgta
      661 ccctataata tgagagttat acattttggt gctggttctg ataaaggagt tgcaccaggt
      721 acagctgttt taagacagtg gttgcctacg ggtacgctgc ttgtcgattc agatcttaat
      781 gactttgtct ctgatgcaga ttcaactttg attggtgatt gtgcaactgt acatacagct
      841 aataaatggg atctcattat tagtgatatg tacgacccta agactaaaaa tgttacaaaa
      901 gaaaatgact ctaaagaggg ttttttcact tacatttgtg ggtttataca acaaaagcta
      961 gctcttggag gttccgtggc tataaagata acagaacatt cttggaatgc tgatctttat
     1021 aagctcatgg gacacttcgc atggtggaca gcctttgtta ctaatgtgaa tgcgtcatca
     1081 tctgaagcat ttttaattgg atgtaattat cttggcaaac cacgcgaaca aatagatggt
     1141 tatgtcatgc atgcaaatta catattttgg aggaatacaa atccaattca gttgtcttcc
     1201 tattctttat ttgacatgag taaatttccc cttaaattaa ggggtactgc tgttatgtct
     1261 ttaaaagaag gtcaaatcaa tgatatgatt ttatctcttc ttagtaaagg tagacttata
     1321 attagagaaa acaacagagt tgttatttct agtgatgttc ttgttaacct cgtgccgaga
     1381 ggttctgatt ataaagatca tgatggtgat tataaagatc atgatattga ttataaagat
     1441 gatgatgata aataaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     1501 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     1561 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     1621 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
     1681 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     1741 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     1801 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     1861 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     1921 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     1981 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     2041 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     2101 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     2161 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     2221 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     2281 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     2341 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     2401 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     2461 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     2521 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     2581 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     2641 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     2701 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca
     2761 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     2821 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     2881 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     2941 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     3001 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     3061 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     3121 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     3181 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     3241 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     3301 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     3361 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     3421 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     3481 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     3541 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     3601 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     3661 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     3721 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac
     3781 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc
     3841 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3901 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3961 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4021 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4081 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4141 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4201 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4261 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4321 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt
     4381 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag
     4441 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca
     4501 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     4561 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     4621 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     4681 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc
     4741 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4801 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4861 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc
     4921 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4981 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5041 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5101 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag
     5161 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5221 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5281 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5341 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc
     5401 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5461 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5521 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc
     5581 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     5641 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact
     5701 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
