LOCUS       sequence-291653-        5617 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 5617)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5617)
  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-m4046613
FEATURES             Location/Qualifiers
     source          1..5617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1372..1390)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1741..2544
                     /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      2627..3609
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3687..3716
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3738..4325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4337..4364)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4388..4579)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4580..5371,5372..5440))
                     /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     5203..5222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5441..5543)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5544..5575)
                     /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 caacaactaa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     1381 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     1441 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     1501 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     1561 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     1621 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     1681 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     1741 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     1801 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     1861 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     1921 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     1981 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     2041 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     2101 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     2161 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     2221 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     2281 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     2341 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     2401 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     2461 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     2521 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     2581 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     2641 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     2701 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     2761 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     2821 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     2881 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     2941 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     3001 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     3061 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     3121 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     3181 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     3241 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     3301 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     3361 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     3421 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     3481 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     3541 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     3601 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag
     3661 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     3721 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3781 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3841 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3901 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3961 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4021 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4081 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4141 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4201 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt
     4261 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     4321 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     4381 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     4441 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     4501 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4561 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct
     4621 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg
     4681 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4741 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa
     4801 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4861 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4921 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4981 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt
     5041 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5101 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5161 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5221 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga
     5281 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5341 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5401 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg
     5461 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     5521 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg
     5581 ctatggaggt caggtatgat ttaaatggtc agttaga
//
