LOCUS       sequence-297893-        6780 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 6780)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6780)
  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-m4046389
FEATURES             Location/Qualifiers
     source          1..6780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1375..1392
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2535..2553)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2904..3707
                     /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      3790..4772
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4850..4879
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4901..5488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5500..5527)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5551..5742)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5743..6534,6535..6603))
                     /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     6366..6385
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6604..6706)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6707..6738)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 tcctctcaag cctggcagcc tggtgtggct atgcccaatt tgtataagat gcaaagaatg
      541 cttctggaga aatgcgattt gcaaaactac ggagacagcg ctacattacc aaagggtatt
      601 atgatgaacg tagcaaagta cacccaattg tgtcaatact taaatacgtt gactttggct
      661 gttccgtata acatgcgtgt catacacttt ggggccggtt cagacaaagg cgtcgctcca
      721 ggtaccgcag ttctaaggca atggctccca actggtactc tattggttga ttctgatctg
      781 aacgacttcg tcagtgatgc tgactccact ttaatcggtg attgtgctac cgttcatacg
      841 gcgaacaagt gggacttgat tatttcggat atgtacgacc caaagaccaa aaatgttaca
      901 aaggaaaatg attctaaaga aggtttcttt acttatatct gtggtttcat ccagcaaaag
      961 cttgccttgg gcggttctgt ggctattaag atcactgaac actcctggaa cgccgactta
     1021 tacaaattga tgggtcattt cgcttggtgg accgcgtttg ttaccaacgt taacgctagt
     1081 tcttcggaag cctttctcat cggttgtaat tatttaggaa agccacgcga acaaattgat
     1141 ggttacgtca tgcacgctaa ctacattttc tggagaaata ctaaccctat acaattgtca
     1201 agctatagtt tattcgacat gtcaaagttt ccattgaagt tgagaggtac agctgtcatg
     1261 tccttgaagg aagggcagat caacgatatg atcttatctt tgctgtctaa aggtagactc
     1321 attattagag aaaataatcg ggttgttatc tcttctgatg tgctagtcaa caacctcgtg
     1381 ccgagaggtt ctaaaatcga ggaaggtaag ctagtcattt ggataaatgg agataagggt
     1441 tacaacggtt tggcagaagt aggcaagaaa tttgaaaagg acacaggtat caaggtgacg
     1501 gttgaacatc cagacaagtt agaagaaaaa ttccctcaag ttgctgctac tggtgatggt
     1561 cccgacatta ttttctgggc ccacgatcgt tttggtggat atgctcagtc aggtttgctg
     1621 gctgagatca ccccagacaa ggccttccaa gataaattgt acccattcac ttgggatgct
     1681 gtcagataca acggtaagct tatagcatat ccaattgctg ttgaagcctt atcgttgatc
     1741 tacaataagg acttgttacc taacccacca aagacatggg aagaaattcc agctttggat
     1801 aaagagctaa aggccaaggg aaaaagcgct ttgatgttta acttacaaga accttatttc
     1861 acctggccat taatcgctgc tgacggtggt tacgcattta aatacgaaaa cggtaagtat
     1921 gacattaagg atgtcggcgt tgataatgct ggtgcaaagg ctggtttgac cttcttggtt
     1981 gatttaataa agaataaaca catgaacgct gacactgact actctatcgc cgaagccgct
     2041 ttcaacaagg gtgaaactgc tatgaccata aatggtccat gggcctggag taacattgat
     2101 acttctaaag taaactatgg tgtcactgtt ttgcctacat tcaaaggcca accatccaag
     2161 ccctttgtcg gtgttctatc cgctggcatc aatgctgctt ctccaaacaa ggagctagct
     2221 aaagaatttt tggaaaatta cttgttaact gacgaaggtt tggaggccgt taacaaggac
     2281 aagccattgg gtgctgtcgc tttgaaatct tacgaagaag aattggctaa ggatcccaga
     2341 attgccgcta ccatggagaa cgcccaaaag ggtgaaatta tgccaaacat cccacaaatg
     2401 tcagccttct ggtacgcagt taggactgcc gtgattaatg ctgcttccgg tagacaaacc
     2461 gtcgacgaag cattaaaaga tgctcaaacg taaacaggcc ccttttcctt tgtcgatatc
     2521 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     2581 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     2641 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     2701 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     2761 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     2821 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     2881 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     2941 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     3001 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     3061 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     3121 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     3181 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     3241 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     3301 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     3361 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     3421 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     3481 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     3541 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     3601 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     3661 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     3721 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     3781 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     3841 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     3901 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     3961 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     4021 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     4081 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     4141 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     4201 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     4261 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     4321 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     4381 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     4441 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     4501 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     4561 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     4621 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     4681 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     4741 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     4801 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     4861 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     4921 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4981 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     5041 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5101 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5161 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5221 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5281 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5341 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     5401 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     5461 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     5521 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     5581 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     5641 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     5701 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     5761 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5821 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     5881 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     5941 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6001 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6061 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6121 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6181 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6241 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     6301 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6361 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6421 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6481 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6541 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6601 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     6661 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     6721 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
