LOCUS       Exported                5664 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 2'-O-ribose 
            methyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046682
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5664)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5664
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     primer_bind     complement(1419..1437)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1788..2591
                     /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      2674..3656
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3734..3763
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3785..4372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4384..4411
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4435..4626)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4627..5487)
                     /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     5250..5269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5488..5590)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5591..5622
                     /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 tcctctcaag cctggcagcc tggtgtggct atgcccaatt tgtataagat gcaaagaatg
      541 cttctggaga aatgcgattt gcaaaactac ggagacagcg ctacattacc aaagggtatt
      601 atgatgaacg tagcaaagta cacccaattg tgtcaatact taaatacgtt gactttggct
      661 gttccgtata acatgcgtgt catacacttt ggggccggtt cagacaaagg cgtcgctcca
      721 ggtaccgcag ttctaaggca atggctccca actggtactc tattggttga ttctgatctg
      781 aacgacttcg tcagtgatgc tgactccact ttaatcggtg attgtgctac cgttcatacg
      841 gcgaacaagt gggacttgat tatttcggat atgtacgacc caaagaccaa aaatgttaca
      901 aaggaaaatg attctaaaga aggtttcttt acttatatct gtggtttcat ccagcaaaag
      961 cttgccttgg gcggttctgt ggctattaag atcactgaac actcctggaa cgccgactta
     1021 tacaaattga tgggtcattt cgcttggtgg accgcgtttg ttaccaacgt taacgctagt
     1081 tcttcggaag cctttctcat cggttgtaat tatttaggaa agccacgcga acaaattgat
     1141 ggttacgtca tgcacgctaa ctacattttc tggagaaata ctaaccctat acaattgtca
     1201 agctatagtt tattcgacat gtcaaagttt ccattgaagt tgagaggtac agctgtcatg
     1261 tccttgaagg aagggcagat caacgatatg atcttatctt tgctgtctaa aggtagactc
     1321 attattagag aaaataatcg ggttgttatc tcttctgatg tgctagtcaa caactaaaca
     1381 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     1441 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     1501 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     1561 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt
     1621 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     1681 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     1741 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta
     1801 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     1861 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     1921 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     1981 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     2041 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     2101 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     2161 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     2221 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     2281 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     2341 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     2401 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt
     2461 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     2521 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     2581 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     2641 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta
     2701 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     2761 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     2821 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     2881 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     2941 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     3001 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     3061 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     3121 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     3181 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     3241 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     3301 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     3361 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     3421 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     3481 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     3541 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     3601 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag
     3661 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg
     3721 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat
     3781 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3841 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg
     3901 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3961 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4021 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4081 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4141 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4201 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4261 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4321 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4381 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag
     4441 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4501 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4561 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4621 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4681 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc
     4741 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4801 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc
     4861 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4921 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4981 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5041 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt
     5101 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg
     5161 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5221 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5281 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt
     5341 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5401 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5461 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa
     5521 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     5581 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag
     5641 gtatgattta aatggtcagt taga
//
