LOCUS       sequence-291480-        6774 bp    DNA     circular SYN 15-APR-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 6774)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6774)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046487
FEATURES             Location/Qualifiers
     source          1..6774
                     /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"
     primer_bind     complement(2529..2547)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2898..3701
                     /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      3784..4766
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4844..4873
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4895..5482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5494..5521)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5545..5736)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5737..6528,6529..6597))
                     /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     6360..6379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6598..6700)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6701..6732)
                     /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 ggttctaaaa tcgaggaagg taagctagtc atttggataa atggagataa gggttacaac
     1441 ggtttggcag aagtaggcaa gaaatttgaa aaggacacag gtatcaaggt gacggttgaa
     1501 catccagaca agttagaaga aaaattccct caagttgctg ctactggtga tggtcccgac
     1561 attattttct gggcccacga tcgttttggt ggatatgctc agtcaggttt gctggctgag
     1621 atcaccccag acaaggcctt ccaagataaa ttgtacccat tcacttggga tgctgtcaga
     1681 tacaacggta agcttatagc atatccaatt gctgttgaag ccttatcgtt gatctacaat
     1741 aaggacttgt tacctaaccc accaaagaca tgggaagaaa ttccagcttt ggataaagag
     1801 ctaaaggcca agggaaaaag cgctttgatg tttaacttac aagaacctta tttcacctgg
     1861 ccattaatcg ctgctgacgg tggttacgca tttaaatacg aaaacggtaa gtatgacatt
     1921 aaggatgtcg gcgttgataa tgctggtgca aaggctggtt tgaccttctt ggttgattta
     1981 ataaagaata aacacatgaa cgctgacact gactactcta tcgccgaagc cgctttcaac
     2041 aagggtgaaa ctgctatgac cataaatggt ccatgggcct ggagtaacat tgatacttct
     2101 aaagtaaact atggtgtcac tgttttgcct acattcaaag gccaaccatc caagcccttt
     2161 gtcggtgttc tatccgctgg catcaatgct gcttctccaa acaaggagct agctaaagaa
     2221 tttttggaaa attacttgtt aactgacgaa ggtttggagg ccgttaacaa ggacaagcca
     2281 ttgggtgctg tcgctttgaa atcttacgaa gaagaattgg ctaaggatcc cagaattgcc
     2341 gctaccatgg agaacgccca aaagggtgaa attatgccaa acatcccaca aatgtcagcc
     2401 ttctggtacg cagttaggac tgccgtgatt aatgctgctt ccggtagaca aaccgtcgac
     2461 gaagcattaa aagatgctca aacgtaaaca ggcccctttt cctttgtcga tatcatgtaa
     2521 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     2581 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     2641 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg gccgccgaac
     2701 cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta atctccgaac
     2761 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     2821 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacgtgca
     2881 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     2941 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     3001 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     3061 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     3121 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac agaaaatttg
     3181 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     3241 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     3301 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     3361 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     3421 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     3481 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat gcattgggtc
     3541 aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt attgttggaa
     3601 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     3661 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     3721 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattacccg
     3781 agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     3841 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     3901 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     3961 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     4021 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     4081 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     4141 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     4201 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     4261 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     4321 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     4381 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     4441 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     4501 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     4561 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     4621 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     4681 gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag
     4741 cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc ggccgcgtgt
     4801 tacaagcttg actccagcgt aactggactg caatcaactc actggctcac cttcacgggt
     4861 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4921 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4981 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5041 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5281 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg cctggtatct
     5401 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     5461 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     5521 caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga aacgcgcgag
     5581 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     5641 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     5701 ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa tcagtgaggc
     5761 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5821 gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5881 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga gggccgagcg
     5941 cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6001 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6061 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6121 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6181 cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6241 ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt actcaaccaa
     6301 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6361 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6421 gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6481 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6541 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatttt
     6601 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     6661 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc ctccggtcgg
     6721 aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt taga
//
