LOCUS       sequence-287727-        5694 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 5694)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5694)
  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-m4046442
FEATURES             Location/Qualifiers
     source          1..5694
                     /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..1404
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1449..1467)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1818..2621
                     /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      2704..3686
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3764..3793
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3815..4402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4414..4441)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4465..4656)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4657..5448,5449..5517))
                     /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     5280..5299
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5518..5620)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5621..5652)
                     /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 ggttctcacc atcatcacca tcattaaaca ggcccctttt cctttgtcga tatcatgtaa
     1441 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     1501 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     1561 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac
     1621 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac
     1681 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     1741 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca
     1801 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     1861 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     1921 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     1981 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     2041 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg
     2101 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     2161 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     2221 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     2281 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     2341 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     2401 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc
     2461 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa
     2521 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     2581 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     2641 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     2701 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     2761 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     2821 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     2881 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     2941 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     3001 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     3061 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     3121 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     3181 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     3241 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     3301 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     3361 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     3421 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     3481 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     3541 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     3601 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag
     3661 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt
     3721 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
     3781 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3841 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3901 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3961 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4021 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4081 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4141 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4201 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4261 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct
     4321 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     4381 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     4441 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag
     4501 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     4561 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     4621 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc
     4681 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4741 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4801 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg
     4861 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4921 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4981 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5041 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5101 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5161 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa
     5221 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5281 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5341 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5401 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5461 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt
     5521 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     5581 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg
     5641 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga
//
