LOCUS       sequence-291624-        5611 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 5611)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5611)
  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-m4046637
FEATURES             Location/Qualifiers
     source          1..5611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1366..1384)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1735..2538
                     /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      2621..3603
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3681..3710
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3732..4319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4331..4358)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4382..4573)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4574..5365,5366..5434))
                     /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     5197..5216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5435..5537)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5538..5569)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgagtcaag cgtggcaacc gggtgttgct atgcctaatc tttacaaaat
      481 gcaaagaatg ctattagaaa agtgtgacct tcaaaattat ggtgatagtg caacattacc
      541 taaaggcata atgatgaatg tcgcaaaata tactcaactg tgtcaatatt taaacacatt
      601 aacattagct gtaccctata atatgagagt tatacatttt ggtgctggtt ctgataaagg
      661 agttgcacca ggtacagctg ttttaagaca gtggttgcct acgggtacgc tgcttgtcga
      721 ttcagatctt aatgactttg tctctgatgc agattcaact ttgattggtg attgtgcaac
      781 tgtacataca gctaataaat gggatctcat tattagtgat atgtacgacc ctaagactaa
      841 aaatgttaca aaagaaaatg actctaaaga gggttttttc acttacattt gtgggtttat
      901 acaacaaaag ctagctcttg gaggttccgt ggctataaag ataacagaac attcttggaa
      961 tgctgatctt tataagctca tgggacactt cgcatggtgg acagcctttg ttactaatgt
     1021 gaatgcgtca tcatctgaag catttttaat tggatgtaat tatcttggca aaccacgcga
     1081 acaaatagat ggttatgtca tgcatgcaaa ttacatattt tggaggaata caaatccaat
     1141 tcagttgtct tcctattctt tatttgacat gagtaaattt ccccttaaat taaggggtac
     1201 tgctgttatg tctttaaaag aaggtcaaat caatgatatg attttatctc ttcttagtaa
     1261 aggtagactt ataattagag aaaacaacag agttgttatt tctagtgatg ttcttgttaa
     1321 ctaaacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     1381 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     1441 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
     1501 atttttcttt tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg
     1561 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     1621 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     1681 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     1741 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     1801 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     1861 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     1921 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     1981 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     2041 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     2101 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     2161 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     2221 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     2281 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     2341 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     2401 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     2461 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     2521 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     2581 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     2641 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     2701 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     2761 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     2821 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     2881 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     2941 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     3001 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     3061 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     3121 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     3181 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     3241 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     3301 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     3361 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     3421 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     3481 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     3541 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     3601 tagctagaga ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac
     3661 tggactgcaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc
     3721 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3781 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg
     3841 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3901 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3961 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4021 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4081 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4141 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4201 gcacgaggga gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4261 acctctgact tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4321 acgccagcaa cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc
     4381 atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     4441 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     4501 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     4561 cctgtttggt cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4621 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct
     4681 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4741 tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat
     4801 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4861 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4921 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4981 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg
     5041 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5101 tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5161 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5221 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac
     5281 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5341 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5401 gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc
     5461 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     5521 ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg
     5581 aggtcaggta tgatttaaat ggtcagttag a
//
