LOCUS       sequence-287707-        5700 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 5700)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5700)
  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-m4046522
FEATURES             Location/Qualifiers
     source          1..5700
                     /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             1369..1386
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1387..1410
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1455..1473)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1824..2627
                     /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      2710..3692
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3770..3799
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3821..4408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4420..4447)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4471..4662)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4663..5454,5455..5523))
                     /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     5286..5305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5524..5626)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5627..5658)
                     /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 ttgttaacct cgtgccgaga
     1381 ggttcttgga gccatccgca gtttgaaaaa taaacaggcc ccttttcctt tgtcgatatc
     1441 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
     1501 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
     1561 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
     1621 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     1681 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     1741 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     1801 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     1861 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     1921 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     1981 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     2041 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     2101 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     2161 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     2221 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     2281 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     2341 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     2401 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     2461 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     2521 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     2581 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     2641 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     2701 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     2761 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     2821 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     2881 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     2941 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     3001 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     3061 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     3121 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     3181 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     3241 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     3301 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     3361 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     3421 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     3481 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     3541 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     3601 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     3661 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     3721 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     3781 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     3841 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3901 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     3961 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4021 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4081 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4141 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4201 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4261 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     4321 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     4381 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     4441 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     4501 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     4561 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     4621 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     4681 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4741 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     4801 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     4861 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4921 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4981 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5041 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5101 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5161 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     5221 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5281 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5341 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5401 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5461 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5521 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     5581 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     5641 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
