LOCUS       sequence-292546-        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-m4046214
FEATURES             Location/Qualifiers
     source          1..5653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..431
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1336..1353
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1354..1377
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1422..1440)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1791..2594
                     /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      2677..3659
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3737..3766
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3788..4375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4387..4414)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4438..4629)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4630..5421,5422..5490))
                     /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     5253..5272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5491..5593)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5594..5625)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aatggtcagt tagagcggcc gccccacaca ccatagcttc aaaatgtttc tactcctttt
       61 ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc
      121 acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa
      181 ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa
      241 aaatttttat cacgtttctt tttcttgaaa attttttttt ttgatttttt tctctttcga
      301 tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca
      361 tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa
      421 tctaatctaa gttttaatta caaaatgagt caagcgtggc aaccgggtgt tgctatgcct
      481 aatctttaca aaatgcaaag aatgctatta gaaaagtgtg accttcaaaa ttatggtgat
      541 agtgcaacat tacctaaagg cataatgatg aatgtcgcaa aatatactca actgtgtcaa
      601 tatttaaaca cattaacatt agctgtaccc tataatatga gagttataca ttttggtgct
      661 ggttctgata aaggagttgc accaggtaca gctgttttaa gacagtggtt gcctacgggt
      721 acgctgcttg tcgattcaga tcttaatgac tttgtctctg atgcagattc aactttgatt
      781 ggtgattgtg caactgtaca tacagctaat aaatgggatc tcattattag tgatatgtac
      841 gaccctaaga ctaaaaatgt tacaaaagaa aatgactcta aagagggttt tttcacttac
      901 atttgtgggt ttatacaaca aaagctagct cttggaggtt ccgtggctat aaagataaca
      961 gaacattctt ggaatgctga tctttataag ctcatgggac acttcgcatg gtggacagcc
     1021 tttgttacta atgtgaatgc gtcatcatct gaagcatttt taattggatg taattatctt
     1081 ggcaaaccac gcgaacaaat agatggttat gtcatgcatg caaattacat attttggagg
     1141 aatacaaatc caattcagtt gtcttcctat tctttatttg acatgagtaa atttcccctt
     1201 aaattaaggg gtactgctgt tatgtcttta aaagaaggtc aaatcaatga tatgatttta
     1261 tctcttctta gtaaaggtag acttataatt agagaaaaca acagagttgt tatttctagt
     1321 gatgttcttg ttaacctcgt gccgagaggt tcttggagcc atccgcagtt tgaaaaataa
     1381 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     1441 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     1501 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt
     1561 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc
     1621 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     1681 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     1741 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag
     1801 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     1861 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     1921 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     1981 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     2041 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     2101 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     2161 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     2221 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     2281 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     2341 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     2401 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     2461 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     2521 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     2581 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     2641 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt
     2701 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2761 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     2821 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     2881 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     2941 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     3001 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     3061 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     3121 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     3181 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     3241 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     3301 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     3361 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     3421 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     3481 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     3541 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     3601 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     3661 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
     3721 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     3781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     3901 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4261 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4321 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4381 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     4441 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     4501 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     4561 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     4621 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4681 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     4741 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4801 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     4861 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4921 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4981 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5041 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     5101 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     5161 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5221 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5281 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     5341 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5401 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5461 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     5521 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     5581 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     5641 caggtatgat tta
//
