LOCUS       sequence-354360-        5694 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 5694)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5694)
  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-m4046442
FEATURES             Location/Qualifiers
     source          1..5694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1345..1362
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1363..1380
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1425..1443)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1794..2597
                     /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      2680..3662
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3740..3769
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3791..4378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4390..4417)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4441..4632)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4633..5424,5425..5493))
                     /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     5256..5275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5494..5596)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5597..5628)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 cggattagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatggaaa agctgcataa ccactttaac taatactttc aacattttca
      361 gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaact ataatgagtc aagcgtggca accgggtgtt
      481 gctatgccta atctttacaa aatgcaaaga atgctattag aaaagtgtga ccttcaaaat
      541 tatggtgata gtgcaacatt acctaaaggc ataatgatga atgtcgcaaa atatactcaa
      601 ctgtgtcaat atttaaacac attaacatta gctgtaccct ataatatgag agttatacat
      661 tttggtgctg gttctgataa aggagttgca ccaggtacag ctgttttaag acagtggttg
      721 cctacgggta cgctgcttgt cgattcagat cttaatgact ttgtctctga tgcagattca
      781 actttgattg gtgattgtgc aactgtacat acagctaata aatgggatct cattattagt
      841 gatatgtacg accctaagac taaaaatgtt acaaaagaaa atgactctaa agagggtttt
      901 ttcacttaca tttgtgggtt tatacaacaa aagctagctc ttggaggttc cgtggctata
      961 aagataacag aacattcttg gaatgctgat ctttataagc tcatgggaca cttcgcatgg
     1021 tggacagcct ttgttactaa tgtgaatgcg tcatcatctg aagcattttt aattggatgt
     1081 aattatcttg gcaaaccacg cgaacaaata gatggttatg tcatgcatgc aaattacata
     1141 ttttggagga atacaaatcc aattcagttg tcttcctatt ctttatttga catgagtaaa
     1201 tttcccctta aattaagggg tactgctgtt atgtctttaa aagaaggtca aatcaatgat
     1261 atgattttat ctcttcttag taaaggtaga cttataatta gagaaaacaa cagagttgtt
     1321 atttctagtg atgttcttgt taacctcgtg ccgagaggtt ctcaccatca tcaccatcat
     1381 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     1441 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     1501 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
     1561 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
     1621 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     1681 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     1741 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     1801 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     1861 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     1921 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     1981 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     2041 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     2101 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     2161 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     2221 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     2281 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     2341 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     2401 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     2461 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     2521 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     2581 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     2641 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     2701 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2761 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     2821 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     2881 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     2941 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     3001 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     3061 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     3121 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     3181 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     3241 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     3301 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     3361 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     3421 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     3481 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     3541 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     3601 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     3661 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     3721 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     3781 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3841 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     3901 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3961 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4021 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4081 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4141 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4201 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4261 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4321 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4381 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     4441 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     4501 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     4561 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     4621 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     4741 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4801 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     4861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     5101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5161 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     5341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5461 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     5521 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     5581 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     5641 ggtcaggtat gatttaaatg gtcagttaga gcggccgctt gctatattga agta
//
