LOCUS       Exported                5718 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3C-like proteinase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046285
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5718)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5718)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5718
                     /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..1428
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1473..1491)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1842..2645
                     /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      2728..3710
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3788..3817
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3839..4426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4438..4465
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4489..4680)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4681..5541)
                     /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     5304..5323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5542..5644)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5645..5676
                     /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 caccatcatc accatcatta aacaggcccc
     1441 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     1501 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     1561 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     1621 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
     1681 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     1741 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     1801 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     1861 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     1921 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     1981 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     2041 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     2101 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     2161 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     2221 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     2281 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     2341 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     2401 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     2461 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     2521 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     2581 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     2641 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     2701 tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa
     2761 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2821 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     2881 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     2941 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     3001 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     3061 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     3121 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     3181 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     3241 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     3301 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     3361 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     3421 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     3481 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     3541 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     3601 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat
     3661 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata
     3721 ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca
     3781 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     3841 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3901 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     3961 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4021 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4081 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4141 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4201 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4261 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4321 tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4381 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4441 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     4501 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     4561 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4621 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4681 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4741 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     4801 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4861 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     4921 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4981 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5041 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5101 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     5161 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     5221 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5281 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5341 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     5401 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5461 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5521 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     5581 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     5641 atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga
     5701 tttaaatggt cagttaga
//
