LOCUS       sequence-298298-        5658 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 5658)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5658)
  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-m4046629
FEATURES             Location/Qualifiers
     source          1..5658
                     /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"
     primer_bind     complement(1413..1431)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1782..2585
                     /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      2668..3650
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3728..3757
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3779..4366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4378..4405)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4429..4620)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4621..5412,5413..5481))
                     /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     5244..5263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5482..5584)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5585..5616)
                     /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 agtcaagcgt ggcaaccggg tgttgctatg cctaatcttt acaaaatgca aagaatgcta
      541 ttagaaaagt gtgaccttca aaattatggt gatagtgcaa cattacctaa aggcataatg
      601 atgaatgtcg caaaatatac tcaactgtgt caatatttaa acacattaac attagctgta
      661 ccctataata tgagagttat acattttggt gctggttctg ataaaggagt tgcaccaggt
      721 acagctgttt taagacagtg gttgcctacg ggtacgctgc ttgtcgattc agatcttaat
      781 gactttgtct ctgatgcaga ttcaactttg attggtgatt gtgcaactgt acatacagct
      841 aataaatggg atctcattat tagtgatatg tacgacccta agactaaaaa tgttacaaaa
      901 gaaaatgact ctaaagaggg ttttttcact tacatttgtg ggtttataca acaaaagcta
      961 gctcttggag gttccgtggc tataaagata acagaacatt cttggaatgc tgatctttat
     1021 aagctcatgg gacacttcgc atggtggaca gcctttgtta ctaatgtgaa tgcgtcatca
     1081 tctgaagcat ttttaattgg atgtaattat cttggcaaac cacgcgaaca aatagatggt
     1141 tatgtcatgc atgcaaatta catattttgg aggaatacaa atccaattca gttgtcttcc
     1201 tattctttat ttgacatgag taaatttccc cttaaattaa ggggtactgc tgttatgtct
     1261 ttaaaagaag gtcaaatcaa tgatatgatt ttatctcttc ttagtaaagg tagacttata
     1321 attagagaaa acaacagagt tgttatttct agtgatgttc ttgttaacta aacaggcccc
     1381 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     1441 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     1501 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     1561 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
     1621 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     1681 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     1741 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     1801 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     1861 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     1921 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     1981 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     2041 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     2101 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     2161 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     2221 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     2281 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     2341 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     2401 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     2461 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     2521 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     2581 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     2641 tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa
     2701 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2761 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     2821 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     2881 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     2941 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     3001 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     3061 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     3121 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     3181 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     3241 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     3301 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     3361 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     3421 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     3481 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     3541 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat
     3601 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata
     3661 ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca
     3721 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     3781 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3841 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     3901 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3961 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4021 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4081 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4141 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4201 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4261 tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4321 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4381 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     4441 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     4501 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4561 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4621 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4681 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     4741 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4801 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     4861 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4921 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4981 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5041 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     5101 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     5161 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5221 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5281 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     5341 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5401 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5461 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     5521 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     5581 atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga
     5641 tttaaatggt cagttaga
//
