LOCUS       Exported                5724 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3C-like proteinase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046200
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5724)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5724)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5724
                     /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"
     CDS             1393..1410
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1411..1434
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1479..1497)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1848..2651
                     /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      2734..3716
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3794..3823
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3845..4432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4444..4471
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4495..4686)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4687..5547)
                     /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     5310..5329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5548..5650)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5651..5682
                     /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 tcctcgtgcc gagaggttct tggagccatc cgcagtttga aaaataaaca
     1441 ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     1501 cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     1561 atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     1621 ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca cggtttcctt
     1681 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac
     1741 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct
     1801 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta
     1861 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
     1921 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
     1981 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
     2041 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
     2101 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     2161 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     2221 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta
     2281 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata
     2341 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     2401 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     2461 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt
     2521 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     2581 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     2641 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     2701 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta
     2761 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     2821 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     2881 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     2941 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     3001 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     3061 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     3121 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     3181 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     3241 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     3301 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     3361 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     3421 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     3481 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     3541 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     3601 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     3661 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag
     3721 agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt aactggactg
     3781 caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa atcaaaggat
     3841 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3901 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aggtaactgg
     3961 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4021 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4081 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4141 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4201 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4261 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4321 ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4381 acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4441 caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc ctcatcagag
     4501 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4561 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4621 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4681 ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4741 atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg gcttaccatc
     4801 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4861 aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt tatccgcctc
     4921 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4981 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5041 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5101 aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt
     5161 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtgagatg
     5221 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5281 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5341 agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct taccgctgtt
     5401 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5461 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5521 ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa atctcgataa
     5581 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     5641 gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta tggaggtcag
     5701 gtatgattta aatggtcagt taga
//
