LOCUS       sequence-287564-        5647 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 5647)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5647)
  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-m4046447
FEATURES             Location/Qualifiers
     source          1..5647
                     /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             1322..1339
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1340..1357
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1402..1420)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1771..2574
                     /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      2657..3639
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3717..3746
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3768..4355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4367..4394)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4418..4609)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4610..5401,5402..5470))
                     /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     5233..5252
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5471..5573)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5574..5605)
                     /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 atgagtcaag cgtggcaacc gggtgttgct atgcctaatc tttacaaaat
      481 gcaaagaatg ctattagaaa agtgtgacct tcaaaattat ggtgatagtg caacattacc
      541 taaaggcata atgatgaatg tcgcaaaata tactcaactg tgtcaatatt taaacacatt
      601 aacattagct gtaccctata atatgagagt tatacatttt ggtgctggtt ctgataaagg
      661 agttgcacca ggtacagctg ttttaagaca gtggttgcct acgggtacgc tgcttgtcga
      721 ttcagatctt aatgactttg tctctgatgc agattcaact ttgattggtg attgtgcaac
      781 tgtacataca gctaataaat gggatctcat tattagtgat atgtacgacc ctaagactaa
      841 aaatgttaca aaagaaaatg actctaaaga gggttttttc acttacattt gtgggtttat
      901 acaacaaaag ctagctcttg gaggttccgt ggctataaag ataacagaac attcttggaa
      961 tgctgatctt tataagctca tgggacactt cgcatggtgg acagcctttg ttactaatgt
     1021 gaatgcgtca tcatctgaag catttttaat tggatgtaat tatcttggca aaccacgcga
     1081 acaaatagat ggttatgtca tgcatgcaaa ttacatattt tggaggaata caaatccaat
     1141 tcagttgtct tcctattctt tatttgacat gagtaaattt ccccttaaat taaggggtac
     1201 tgctgttatg tctttaaaag aaggtcaaat caatgatatg attttatctc ttcttagtaa
     1261 aggtagactt ataattagag aaaacaacag agttgttatt tctagtgatg ttcttgttaa
     1321 cctcgtgccg agaggttctc accatcatca ccatcattaa acaggcccct tttcctttgt
     1381 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
     1441 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
     1501 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
     1561 gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
     1621 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     1681 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     1741 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     1801 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     1861 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     1921 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     1981 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     2041 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     2101 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     2161 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     2221 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     2281 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     2341 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     2401 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     2461 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     2521 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     2581 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     2641 cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     2701 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2761 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     2821 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2881 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     2941 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     3001 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     3061 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     3121 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     3181 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     3241 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     3301 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     3361 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     3421 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     3481 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     3541 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt
     3601 tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg
     3661 agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct
     3721 caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt
     3781 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3841 ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga
     3901 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3961 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4021 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4081 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4141 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4201 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa
     4261 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt
     4321 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca
     4381 cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac
     4441 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     4501 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     4561 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct
     4621 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4681 tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa
     4741 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4801 ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt
     4861 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4921 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4981 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5041 tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5101 cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg
     5161 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5221 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5281 aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5341 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5401 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5461 gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     5521 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa
     5581 agcctccggt cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc
     5641 agttaga
//
