LOCUS       sequence-300403-        6733 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 6733)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6733)
  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-m4046279
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(299..1102)
                     /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"
     primer_bind     1453..1471
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2614..2631)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(3542..3955)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      4001..4032
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        4033..4135
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4136..4204,4205..4996)
                     /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     complement(4354..4373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4997..5188
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      5212..5239
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(5251..5838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5860..5889)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(5967..6733,1..216)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
       61 gttttacaaa aatgaagcac agattcttcg ttggtaaaat agcgctttcg cgttgcattt
      121 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca
      181 tttttgttct acaaaatgaa gcacagatgc ttcgttactc gggtaataac tgatataatt
      241 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
      301 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
      361 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
      421 atcctgtaga aaccacatca tccacggttc tatactgttg acccaatgca tctcccttgt
      481 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
      541 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
      601 tacccttagt atattctcca gtagataggg agcccttgca tgacaattct gctaacatca
      661 aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac
      721 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
      781 ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcctcgaaga
      841 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
      901 aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
      961 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
     1021 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
     1081 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcacgttttt gttctgtgca
     1141 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
     1201 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     1261 aaaaatttca aggaaaccgt gtactgcagt gcactagtgg ttcggcggcc gctttgtaca
     1321 gaaaaaaaag aaaaatttga aatataaata acgttcttaa tactaacata actattaaaa
     1381 aaaataaata gggacctaga cttcaggttg tctaactcct tccttttcgg ttagagcgga
     1441 tgtgggagga gggcgtgaat gtaagcgtga cataactaat tacatgatat cgacaaagga
     1501 aaaggggcct gtttacgttt gagcatcttt taatgcttcg tcgacggttt gtctaccgga
     1561 agcagcatta atcacggcag tcctaactgc gtaccagaag gctgacattt gtgggatgtt
     1621 tggcataatt tcaccctttt gggcgttctc catggtagcg gcaattctgg gatccttagc
     1681 caattcttct tcgtaagatt tcaaagcgac agcacccaat ggcttgtcct tgttaacggc
     1741 ctccaaacct tcgtcagtta acaagtaatt ttccaaaaat tctttagcta gctccttgtt
     1801 tggagaagca gcattgatgc cagcggatag aacaccgaca aagggcttgg atggttggcc
     1861 tttgaatgta ggcaaaacag tgacaccata gtttacttta gaagtatcaa tgttactcca
     1921 ggcccatgga ccatttatgg tcatagcagt ttcacccttg ttgaaagcgg cttcggcgat
     1981 agagtagtca gtgtcagcgt tcatgtgttt attctttatt aaatcaacca agaaggtcaa
     2041 accagccttt gcaccagcat tatcaacgcc gacatcctta atgtcatact taccgttttc
     2101 gtatttaaat gcgtaaccac cgtcagcagc gattaatggc caggtgaaat aaggttcttg
     2161 taagttaaac atcaaagcgc tttttccctt ggcctttagc tctttatcca aagctggaat
     2221 ttcttcccat gtctttggtg ggttaggtaa caagtcctta ttgtagatca acgataaggc
     2281 ttcaacagca attggatatg ctataagctt accgttgtat ctgacagcat cccaagtgaa
     2341 tgggtacaat ttatcttgga aggccttgtc tggggtgatc tcagccagca aacctgactg
     2401 agcatatcca ccaaaacgat cgtgggccca gaaaataatg tcgggaccat caccagtagc
     2461 agcaacttga gggaattttt cttctaactt gtctggatgt tcaaccgtca ccttgatacc
     2521 tgtgtccttt tcaaatttct tgcctacttc tgccaaaccg ttgtaaccct tatctccatt
     2581 tatccaaatg actagcttac cttcctcgat tttagaacct ctcggcacga ggttgttgac
     2641 tagcacatca gaagagataa caacccgatt attttctcta ataatgagtc tacctttaga
     2701 cagcaaagat aagatcatat cgttgatctg cccttccttc aaggacatga cagctgtacc
     2761 tctcaacttc aatggaaact ttgacatgtc gaataaacta tagcttgaca attgtatagg
     2821 gttagtattt ctccagaaaa tgtagttagc gtgcatgacg taaccatcaa tttgttcgcg
     2881 tggctttcct aaataattac aaccgatgag aaaggcttcc gaagaactag cgttaacgtt
     2941 ggtaacaaac gcggtccacc aagcgaaatg acccatcaat ttgtataagt cggcgttcca
     3001 ggagtgttca gtgatcttaa tagccacaga accgcccaag gcaagctttt gctggatgaa
     3061 accacagata taagtaaaga aaccttcttt agaatcattt tcctttgtaa catttttggt
     3121 ctttgggtcg tacatatccg aaataatcaa gtcccacttg ttcgccgtat gaacggtagc
     3181 acaatcaccg attaaagtgg agtcagcatc actgacgaag tcgttcagat cagaatcaac
     3241 caatagagta ccagttggga gccattgcct tagaactgcg gtacctggag cgacgccttt
     3301 gtctgaaccg gccccaaagt gtatgacacg catgttatac ggaacagcca aagtcaacgt
     3361 atttaagtat tgacacaatt gggtgtactt tgctacgttc atcataatac cctttggtaa
     3421 tgtagcgctg tctccgtagt tttgcaaatc gcatttctcc agaagcattc tttgcatctt
     3481 atacaaattg ggcatagcca caccaggctg ccaggcttga gaggacattt tgtaattaaa
     3541 acttagatta gattgctatg ctttctttct aatgagcaag aagtaaaaaa agttgtaata
     3601 gaacaagaaa aatgaaactg aaacttgaga aattgaagac cgtttattaa cttaaatatc
     3661 aatgggaggt catcgaaaga gaaaaaaatc aaaaaaaaaa attttcaaga aaaagaaacg
     3721 tgataaaaat ttttattgcc tttttcgacg aagaaaaaga aacgaggcgg tctctttttt
     3781 cttttccaaa cctttagtac gggtaattaa cgacacccta gaggaagaaa gaggggaaat
     3841 ttagtatgct gtgcttgggt gttttgaagt ggtacggcga tgcgcggagt ccgagaaaat
     3901 ctggaagagt aaaaaaggag tagaaacatt ttgaagctat ggtgtgtggg gcggccgctc
     3961 taactgacca tttaaatcat acctgacctc catagcagaa agtcaaaagc ctccgaccgg
     4021 aggcttttga cttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     4081 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgag
     4141 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4201 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4261 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagtttac gccccgaaga
     4321 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4381 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4441 gtactcacca gtcacagaaa agcatctcac ggatggcatg acagtaagag aattatgcag
     4501 tgctgccata accatgagtg ataacactgc ggccaactta cttctggcaa cgatcggagg
     4561 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4621 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4681 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4741 gcaacaatta atagactgga tggaggcgga taaagttgca ggatcacttc tgcgctcggc
     4801 cctcccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4861 tatcattgca gcactggggc cagatggtaa gccctcccgc atcgtagtta tctacacgac
     4921 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4981 gattaagcat tggtaagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
     5041 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
     5101 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
     5161 tttcggtgat gacggtgaaa acctctgatg agggcccaaa tgtaatcacc tggctcacct
     5221 tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5281 tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5341 ggcgtttacc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5401 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5461 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5521 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5581 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5641 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5701 tggtatctgc gctctgctga agccagttac ctcggaaaaa gagttggtag ctcttgatcc
     5761 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5821 agaaaaaaag gatctcaaga agatcctttg attttctacc gaagaaaggc ccacccgtga
     5881 aggtgagcca gtgagttgat tgcagtccag ttacgctgga gtcaagcttg taacacgcgg
     5941 ccgctcggga agttcctatt ctctagctag aaagtatagg aacttcagag cgcttttgaa
     6001 aaccaaaagc gctctgatga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga
     6061 gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta
     6121 tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg
     6181 catctaaact cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa
     6241 aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt
     6301 cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga
     6361 agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga
     6421 atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt
     6481 aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc
     6541 ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc
     6601 gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat
     6661 gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg
     6721 ttgcatttct gtt
//
