LOCUS       sequence-323027-        5647 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 5647)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5647)
  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-m4046447
FEATURES             Location/Qualifiers
     source          1..5647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..408
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1313..1330
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1331..1348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1393..1411)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1762..2565
                     /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      2648..3630
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3708..3737
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3759..4346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4358..4385)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4409..4600)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4601..5392,5393..5461))
                     /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     5224..5243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5462..5564)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5565..5596)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga
       61 ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct
      121 ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac
      181 cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt
      241 cttgaaaatt tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt
      301 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
      361 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa
      421 aatgagtcaa gcgtggcaac cgggtgttgc tatgcctaat ctttacaaaa tgcaaagaat
      481 gctattagaa aagtgtgacc ttcaaaatta tggtgatagt gcaacattac ctaaaggcat
      541 aatgatgaat gtcgcaaaat atactcaact gtgtcaatat ttaaacacat taacattagc
      601 tgtaccctat aatatgagag ttatacattt tggtgctggt tctgataaag gagttgcacc
      661 aggtacagct gttttaagac agtggttgcc tacgggtacg ctgcttgtcg attcagatct
      721 taatgacttt gtctctgatg cagattcaac tttgattggt gattgtgcaa ctgtacatac
      781 agctaataaa tgggatctca ttattagtga tatgtacgac cctaagacta aaaatgttac
      841 aaaagaaaat gactctaaag agggtttttt cacttacatt tgtgggttta tacaacaaaa
      901 gctagctctt ggaggttccg tggctataaa gataacagaa cattcttgga atgctgatct
      961 ttataagctc atgggacact tcgcatggtg gacagccttt gttactaatg tgaatgcgtc
     1021 atcatctgaa gcatttttaa ttggatgtaa ttatcttggc aaaccacgcg aacaaataga
     1081 tggttatgtc atgcatgcaa attacatatt ttggaggaat acaaatccaa ttcagttgtc
     1141 ttcctattct ttatttgaca tgagtaaatt tccccttaaa ttaaggggta ctgctgttat
     1201 gtctttaaaa gaaggtcaaa tcaatgatat gattttatct cttcttagta aaggtagact
     1261 tataattaga gaaaacaaca gagttgttat ttctagtgat gttcttgtta acctcgtgcc
     1321 gagaggttct caccatcatc accatcatta aacaggcccc ttttcctttg tcgatatcat
     1381 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa
     1441 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta
     1501 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc
     1561 gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc
     1621 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     1681 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg
     1741 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     1801 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     1861 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     1921 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     1981 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa
     2041 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     2101 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     2161 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     2221 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     2281 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     2341 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg
     2401 ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt
     2461 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     2521 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     2581 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     2641 cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
     2701 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
     2761 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
     2821 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     2881 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
     2941 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
     3001 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
     3061 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
     3121 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
     3181 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
     3241 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
     3301 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
     3361 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
     3421 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
     3481 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
     3541 tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca
     3601 aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc
     3661 gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac
     3721 gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc
     3781 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3841 caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata
     3901 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3961 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4021 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4081 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4141 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4201 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt
     4261 atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct
     4321 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt
     4381 gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg
     4441 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     4501 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     4561 agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg
     4621 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4681 tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc
     4741 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg
     4801 agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4861 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4921 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4981 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5041 cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5101 ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa
     5161 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5221 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5281 cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5341 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5401 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5461 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     5521 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg
     5581 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagttagagc
     5641 ggccgcc
//
