LOCUS       sequence-298135-        5653 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 5653)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5653)
  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-m4046191
FEATURES             Location/Qualifiers
     source          1..5653
                     /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..1363
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1408..1426)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1777..2580
                     /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      2663..3645
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3723..3752
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3774..4361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4373..4400)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4424..4615)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4616..5407,5408..5476))
                     /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     5239..5258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5477..5579)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5580..5611)
                     /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 ntttatcacg
      241 tttnttnnnn nnnaaaattt 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 gttctcacca tcatcaccat cattaaacag gccccttttc
     1381 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     1441 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1501 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     1561 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     1621 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     1681 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     1741 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     1801 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     1861 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     1921 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     1981 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     2041 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     2101 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     2161 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     2221 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     2281 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     2341 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     2401 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     2461 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     2521 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     2581 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     2641 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     2701 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     2761 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     2821 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2881 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     2941 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     3001 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     3061 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     3121 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     3181 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     3241 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     3301 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     3361 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     3421 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     3481 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     3541 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     3601 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     3661 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     3721 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     3781 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3841 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     3901 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3961 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4021 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4081 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4141 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4201 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4261 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     4321 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     4381 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     4441 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     4501 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     4561 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     4621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4681 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     4741 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4801 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     4861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4921 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5041 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     5101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     5161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5221 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5281 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     5341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5461 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     5521 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     5581 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     5641 atggtcagtt aga
//
