LOCUS       sequence-297875-        5701 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 5701)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5701)
  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-m4046265
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1328..1345
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1346..1411
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1456..1474)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1825..2628
                     /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      2711..3693
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3771..3800
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3822..4409
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4421..4448)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4472..4663)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4664..5455,5456..5524))
                     /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     5287..5306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5525..5627)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5628..5659)
                     /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 atgtcctctc aagcctggca gcctggtgtg gctatgccca atttgtataa
      481 gatgcaaaga atgcttctgg agaaatgcga tttgcaaaac tacggagaca gcgctacatt
      541 accaaagggt attatgatga acgtagcaaa gtacacccaa ttgtgtcaat acttaaatac
      601 gttgactttg gctgttccgt ataacatgcg tgtcatacac tttggggccg gttcagacaa
      661 aggcgtcgct ccaggtaccg cagttctaag gcaatggctc ccaactggta ctctattggt
      721 tgattctgat ctgaacgact tcgtcagtga tgctgactcc actttaatcg gtgattgtgc
      781 taccgttcat acggcgaaca agtgggactt gattatttcg gatatgtacg acccaaagac
      841 caaaaatgtt acaaaggaaa atgattctaa agaaggtttc tttacttata tctgtggttt
      901 catccagcaa aagcttgcct tgggcggttc tgtggctatt aagatcactg aacactcctg
      961 gaacgccgac ttatacaaat tgatgggtca tttcgcttgg tggaccgcgt ttgttaccaa
     1021 cgttaacgct agttcttcgg aagcctttct catcggttgt aattatttag gaaagccacg
     1081 cgaacaaatt gatggttacg tcatgcacgc taactacatt ttctggagaa atactaaccc
     1141 tatacaattg tcaagctata gtttattcga catgtcaaag tttccattga agttgagagg
     1201 tacagctgtc atgtccttga aggaagggca gatcaacgat atgatcttat ctttgctgtc
     1261 taaaggtaga ctcattatta gagaaaataa tcgggttgtt atctcttctg atgtgctagt
     1321 caacaacctc gtgccgagag gttctgatta taaagatcat gatggtgatt ataaagatca
     1381 tgatattgat tataaagatg atgatgataa ataaacaggc cccttttcct ttgtcgatat
     1441 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
     1501 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
     1561 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
     1621 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     1681 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     1741 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     1801 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     1861 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     1921 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     1981 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     2041 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     2101 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     2161 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     2221 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     2281 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     2341 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     2401 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     2461 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     2521 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     2581 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     2641 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     2701 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     2761 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     2821 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     2881 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2941 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     3001 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     3061 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     3121 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     3181 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     3241 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     3301 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     3361 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     3421 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     3481 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     3541 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     3601 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     3661 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     3721 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     3781 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     3841 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3901 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     3961 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4021 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4081 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4141 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4201 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4261 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     4321 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     4381 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     4441 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     4501 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     4561 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     4621 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     4681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4741 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     4801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     4861 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5101 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5161 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     5221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5341 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5521 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     5581 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     5641 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     5701 a
//
