LOCUS       Exported                5688 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3C-like proteinase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046679
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5688)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5688)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5688
                     /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(1443..1461)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1812..2615
                     /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      2698..3680
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3758..3787
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3809..4396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4408..4435
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4459..4650)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4651..5511)
                     /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     5274..5293
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5512..5614)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5615..5646
                     /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 tcgggtttca gaaaaatggc ctttcctagt ggaaaggtcg agggctgtat ggtgcaagta
      541 acatgcggta cgaccacttt gaacggttta tggcttgacg atgttgttta ctgtccacgt
      601 catgtcatct gtacttcaga agatatgttg aatccaaact atgaagacct actgattcgc
      661 aagtctaacc acaatttctt agttcaggct ggtaacgttc aattgagagt tataggtcac
      721 agcatgcaaa attgtgtctt gaagttaaaa gttgataccg caaacccgaa gactccaaag
      781 tacaaatttg tcagaatcca acccggtcaa accttctccg tcttggcttg ttataacggg
      841 tccccatctg gtgtatacca gtgcgctatg aggccaaact tcactattaa gggttctttt
      901 ttgaatggtt cttgtggctc cgttggtttt aatatcgact acgattgcgt cagtttctgt
      961 tacatgcatc acatggaatt gcctacaggt gttcacgccg gaaccgacct agaagggaac
     1021 ttctatggtc cattcgttga tcggcaaact gctcaagctg cgggtacgga taccacaatt
     1081 actgtaaatg tgcttgcctg gttgtacgct gctgttataa acggtgaccg atggttttta
     1141 aacagattca ctaccacatt gaatgacttt aatctggtcg ctatgaaata taactacgaa
     1201 ccactcactc aagatcatgt cgatatttta ggtcctttgt ctgcacaaac tggcatcgct
     1261 gttttggaca tgtgtgcgtc ccttaaggaa ctactccaaa acggtatgaa cgggagaacg
     1321 attttgggtt cagccctatt ggaggacgaa tttaccccat tcgatgtggt tagacagtgc
     1381 tctggagtaa ccttccaata aacaggcccc ttttcctttg tcgatatcat gtaattagtt
     1441 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
     1501 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
     1561 cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc gaaccactag
     1621 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     1681 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     1741 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     1801 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     1861 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     1921 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     1981 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     2041 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     2101 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     2161 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     2221 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     2281 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     2341 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     2401 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     2461 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     2521 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     2581 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     2641 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaac
     2701 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     2761 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     2821 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     2881 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     2941 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     3001 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     3061 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     3121 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     3181 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     3241 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     3301 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     3361 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     3421 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     3481 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     3541 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     3601 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     3661 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaag
     3721 cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     3781 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3901 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3961 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4021 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4081 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4141 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4201 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4261 ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt atctttatag
     4321 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     4381 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     4441 attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4501 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4561 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4621 gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat
     4681 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4741 tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4801 ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag
     4861 tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4921 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4981 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5041 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc
     5101 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5161 tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5221 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5281 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa
     5341 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5401 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5461 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc
     5521 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     5581 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     5641 ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
