LOCUS       sequence-287636-        5659 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 5659)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5659)
  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-m4046161
FEATURES             Location/Qualifiers
     source          1..5659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1328..1345
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1346..1369
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1414..1432)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1783..2586
                     /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      2669..3651
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3729..3758
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3780..4367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4379..4406)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4430..4621)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4622..5413,5414..5482))
                     /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     5245..5264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5483..5585)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5586..5617)
                     /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 tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgtcctctc aagcctggca gcctggtgtg gctatgccca atttgtataa
      481 gatgcaaaga atgcttctgg agaaatgcga tttgcaaaac tacggagaca gcgctacatt
      541 accaaagggt attatgatga acgtagcaaa gtacacccaa ttgtgtcaat acttaaatac
      601 gttgactttg gctgttccgt ataacatgcg tgtcatacac tttggggccg gttcagacaa
      661 aggcgtcgct ccaggtaccg cagttctaag gcaatggctc ccaactggta ctctattggt
      721 tgattctgat ctgaacgact tcgtcagtga tgctgactcc actttaatcg gtgattgtgc
      781 taccgttcat acggcgaaca agtgggactt gattatttcg gatatgtacg acccaaagac
      841 caaaaatgtt acaaaggaaa atgattctaa agaaggtttc tttacttata tctgtggttt
      901 catccagcaa aagcttgcct tgggcggttc tgtggctatt aagatcactg aacactcctg
      961 gaacgccgac ttatacaaat tgatgggtca tttcgcttgg tggaccgcgt ttgttaccaa
     1021 cgttaacgct agttcttcgg aagcctttct catcggttgt aattatttag gaaagccacg
     1081 cgaacaaatt gatggttacg tcatgcacgc taactacatt ttctggagaa atactaaccc
     1141 tatacaattg tcaagctata gtttattcga catgtcaaag tttccattga agttgagagg
     1201 tacagctgtc atgtccttga aggaagggca gatcaacgat atgatcttat ctttgctgtc
     1261 taaaggtaga ctcattatta gagaaaataa tcgggttgtt atctcttctg atgtgctagt
     1321 caacaacctc gtgccgagag gttcttggag ccatccgcag tttgaaaaat aaacaggccc
     1381 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
     1441 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     1501 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1561 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     1621 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     1681 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     1741 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     1801 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     1861 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     1921 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     1981 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     2041 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     2101 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     2161 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     2221 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     2281 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     2341 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     2401 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     2461 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     2521 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     2581 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     2641 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     2701 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     2761 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     2821 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     2881 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     2941 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     3001 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     3061 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     3121 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     3181 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     3241 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     3301 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     3361 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     3421 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     3481 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     3541 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     3601 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     3661 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     3721 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     3781 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3841 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     3901 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3961 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4021 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4081 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4141 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4201 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4261 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4321 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4381 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     4441 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4501 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4561 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4621 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4681 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     4741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4801 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     4861 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4921 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5041 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     5101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     5161 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5281 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     5341 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5461 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     5521 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     5581 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     5641 atttaaatgg tcagttaga
//
