LOCUS       sequence-298294-        5584 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 2'-O-ribose 
            methyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5584)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5584)
  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..5584
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(1339..1357)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1708..2511
                     /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
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWXAYLRRCGQQN"
     rep_origin      2594..3576
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3654..3683
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3705..4292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4304..4331)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4355..4546)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4547..5338,5339..5407))
                     /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     5170..5189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5408..5510)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5511..5542)
                     /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 cgtttctttt tcttgaaaat tttttttttt
      241 ngattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat
      301 ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat
      361 tagaaagaaa gcatagcaat ctaatctaag ttttaattac aaaatgagtc aagcgtggca
      421 accgggtgtt gctatgccta atctttacaa aatgcaaaga atgctattag aaaagtgtga
      481 ccttcaaaat tatggtgata gtgcaacatt acctaaaggc ataatgatga atgtcgcaaa
      541 atatactcaa ctgtgtcaat atttaaacac attaacatta gctgtaccct ataatatgag
      601 agttatacat tttggtgctg gttctgataa aggagttgca ccaggtacag ctgttttaag
      661 acagtggttg cctacgggta cgctgcttgt cgattcagat cttaatgact ttgtctctga
      721 tgcagattca actttgattg gtgattgtgc aactgtacat acagctaata aatgggatct
      781 cattattagt gatatgtacg accctaagac taaaaatgtt acaaaagaaa atgactctaa
      841 agagggtttt ttcacttaca tttgtgggtt tatacaacaa aagctagctc ttggaggttc
      901 cgtggctata aagataacag aacattcttg gaatgctgat ctttataagc tcatgggaca
      961 cttcgcatgg tggacagcct ttgttactaa tgtgaatgcg tcatcatctg aagcattttt
     1021 aattggatgt aattatcttg gcaaaccacg cgaacaaata gatggttatg tcatgcatgc
     1081 aaattacata ttttggagga atacaaatcc aattcagttg tcttcctatt ctttatttga
     1141 catgagtaaa tttcccctta aattaagggg tactgctgtt atgtctttaa aagaaggtca
     1201 aatcaatgat atgattttat ctcttcttag taaaggtaga cttataatta gagaaaacaa
     1261 cagagttgtt atttctagtg atgttcttgt taactaaaca ggcccctttt cctttgtcga
     1321 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac
     1381 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt
     1441 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg
     1501 gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta
     1561 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     1621 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     1681 aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct
     1741 actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac
     1801 ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta
     1861 ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag
     1921 ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac
     1981 agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga
     2041 atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc
     2101 ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca
     2161 gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt
     2221 gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga
     2281 gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat
     2341 gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt
     2401 attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     2461 agaaaagcag gctgggangc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     2521 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     2581 ttattacccg agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     2641 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     2701 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     2761 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2821 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     2881 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     2941 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     3001 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     3061 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     3121 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     3181 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     3241 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     3301 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     3361 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     3421 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     3481 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg
     3541 ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc
     3601 ggccgcgtgt tacaagcttg actccagcgt aactggactg caatcaactc actggctcac
     3661 cttcacgggt gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt
     3721 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3781 ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac
     3841 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3901 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3961 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4021 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4081 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4141 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg
     4201 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt
     4261 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc
     4321 gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga
     4381 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     4441 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4501 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa
     4561 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4621 ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga
     4681 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga
     4741 gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt
     4801 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4861 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4921 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4981 gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5041 cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt
     5101 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5161 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5221 gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5281 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5341 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5401 tactcatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     5461 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc
     5521 ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt
     5581 taga
//
