LOCUS       sequence-287739-        5748 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 5748)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5748)
  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-m4046554
FEATURES             Location/Qualifiers
     source          1..5748
                     /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..1458
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1503..1521)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1872..2675
                     /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      2758..3740
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3818..3847
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3869..4456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4468..4495)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4519..4710)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4711..5502,5503..5571))
                     /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     5334..5353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5572..5674)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5675..5706)
                     /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 ctgattataa agatcatgat ggtgattata aagatcatga tattgattat
     1441 aaagatgatg atgataaata aacaggcccc ttttcctttg tcgatatcat gtaattagtt
     1501 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
     1561 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
     1621 cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc gaaccactag
     1681 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     1741 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     1801 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     1861 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     1921 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     1981 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     2041 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     2101 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     2161 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     2221 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     2281 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     2341 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     2401 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     2461 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     2521 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     2581 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     2641 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     2701 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaac
     2761 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     2821 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     2881 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     2941 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     3001 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     3061 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     3121 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     3181 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     3241 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     3301 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     3361 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     3421 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     3481 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     3541 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     3601 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     3661 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     3721 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaag
     3781 cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     3841 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3901 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3961 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4021 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4081 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4141 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4201 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4261 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4321 ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt atctttatag
     4381 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     4441 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     4501 attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4561 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4621 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4681 gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat
     4741 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4801 tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4861 ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag
     4921 tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4981 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5041 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5101 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc
     5161 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5221 tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5281 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5341 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa
     5401 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5461 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5521 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc
     5581 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     5641 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     5701 ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
