LOCUS       Exported                5682 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast expression plasmid for SARS-CoV-2 3C-like proteinase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046647
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5682)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5682
                     /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(1437..1455)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1806..2609
                     /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      2692..3674
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3752..3781
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3803..4390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4402..4429
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4453..4644)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4645..5505)
                     /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     5268..5287
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5506..5608)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5609..5640
                     /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 ggttttagaa aaatggcatt cccatctggt aaagttgagg gttgtatggt acaagtaact
      541 tgtggtacaa ctacacttaa cggtctttgg cttgatgacg tagtttactg tccaagacat
      601 gtgatctgca cctctgaaga catgcttaac cctaattatg aagatttact cattcgtaag
      661 tctaatcata atttcttggt acaggctggt aatgttcaac tcagggttat tggacattct
      721 atgcaaaatt gtgtacttaa gcttaaggtt gatacagcca atcctaagac acctaagtat
      781 aagtttgttc gcattcaacc aggacagact ttttcagtgt tagcttgtta caatggttca
      841 ccatctggtg tttaccaatg tgctatgagg cccaatttca ctattaaggg ttcattcctt
      901 aatggttcat gtggtagtgt tggttttaac atagattatg actgtgtctc tttttgttac
      961 atgcaccata tggaattacc aactggagtt catgctggca cagacttaga aggtaacttt
     1021 tatggacctt ttgttgacag gcaaacagca caagcagctg gtacggacac aactattaca
     1081 gttaatgttt tagcttggtt gtacgctgct gttataaatg gagacaggtg gtttctcaat
     1141 cgatttacca caactcttaa tgactttaac cttgtggcta tgaagtacaa ttatgaacct
     1201 ctaacacaag accatgttga catactagga cctctttctg ctcaaactgg aattgccgtt
     1261 ttagatatgt gtgcttcatt aaaagaatta ctgcaaaatg gtatgaatgg acgtaccata
     1321 ttgggtagtg ctttattaga agatgaattt acaccttttg atgttgttag acaatgctca
     1381 ggtgttactt tctaaacagg ccccttttcc tttgtcgata tcatgtaatt agttatgtca
     1441 cgcttacatt cacgccctcc tcccacatcc gctctaaccg aaaaggaagg agttagacaa
     1501 cctgaagtct aggtccctat ttattttttt taatagttat gttagtatta agaacgttat
     1561 ttatatttca aatttttctt ttttttctgt acaaagcggc cgccgaacca ctagtgcact
     1621 gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     1681 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     1741 gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg aaacgaagat
     1801 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     1861 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     1921 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     1981 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     2041 cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta
     2101 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     2161 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     2221 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     2281 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     2341 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     2401 ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa cagtatagaa
     2461 ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga ggactatttg
     2521 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     2581 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     2641 taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag taacgaagca
     2701 tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     2761 aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga
     2821 agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa
     2881 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     2941 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     3001 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     3061 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     3121 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     3181 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     3241 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     3301 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     3361 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     3421 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     3481 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     3541 gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt
     3601 tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt
     3661 cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta caagcttgac
     3721 tccagcgtaa ctggactgca atcaactcac tggctcacct tcacgggtgg gcctttcttc
     3781 ggtagaaaat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3841 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3901 tttttccgag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3961 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4021 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4081 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4141 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4201 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4261 aacaggagag cgcacgaggg agcttccagg ggtaaacgcc tggtatcttt atagtcctgt
     4321 cgggtttcgc cacctctgac ttgagcatcg atttttgtga tgctcgtcag gggggcggag
     4381 cctatggaaa aacgccagca acgcagaaag gcccacccga aggtgagcca ggtgattaca
     4441 tttgggccct catcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     4501 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     4561 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     4621 ggcggttttt tcctgtttgg tcacttacca atgcttaatc agtgaggcac ctatctcagc
     4681 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4741 gcgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4801 ggctccagat ttatcagcaa taaaccagcc agccgggagg gccgagcgca gaagtgatcc
     4861 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4921 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4981 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5041 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgccagaag
     5101 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5161 catgccatcc gtgagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5221 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5281 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gtaaactctc
     5341 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5401 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5461 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcattttag cttccttagc
     5521 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     5581 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact
     5641 ttctgctatg gaggtcaggt atgatttaaa tggtcagtta ga
//
