LOCUS       Exported                5719 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3C-like proteinase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046511
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5719)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5719)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5719
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1346..1363
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1364..1429
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1474..1492)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1843..2646
                     /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      2729..3711
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3789..3818
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3840..4427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4439..4466
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4490..4681)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4682..5542)
                     /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     5305..5324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5543..5645)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5646..5677
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgggtttta gaaaaatggc attcccatct ggtaaagttg agggttgtat
      481 ggtacaagta acttgtggta caactacact taacggtctt tggcttgatg acgtagttta
      541 ctgtccaaga catgtgatct gcacctctga agacatgctt aaccctaatt atgaagattt
      601 actcattcgt aagtctaatc ataatttctt ggtacaggct ggtaatgttc aactcagggt
      661 tattggacat tctatgcaaa attgtgtact taagcttaag gttgatacag ccaatcctaa
      721 gacacctaag tataagtttg ttcgcattca accaggacag actttttcag tgttagcttg
      781 ttacaatggt tcaccatctg gtgtttacca atgtgctatg aggcccaatt tcactattaa
      841 gggttcattc cttaatggtt catgtggtag tgttggtttt aacatagatt atgactgtgt
      901 ctctttttgt tacatgcacc atatggaatt accaactgga gttcatgctg gcacagactt
      961 agaaggtaac ttttatggac cttttgttga caggcaaaca gcacaagcag ctggtacgga
     1021 cacaactatt acagttaatg ttttagcttg gttgtacgct gctgttataa atggagacag
     1081 gtggtttctc aatcgattta ccacaactct taatgacttt aaccttgtgg ctatgaagta
     1141 caattatgaa cctctaacac aagaccatgt tgacatacta ggacctcttt ctgctcaaac
     1201 tggaattgcc gttttagata tgtgtgcttc attaaaagaa ttactgcaaa atggtatgaa
     1261 tggacgtacc atattgggta gtgctttatt agaagatgaa tttacacctt ttgatgttgt
     1321 tagacaatgc tcaggtgtta ctttcctcgt gccgagaggt tctgattata aagatcatga
     1381 tggtgattat aaagatcatg atattgatta taaagatgat gatgataaat aaacaggccc
     1441 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
     1501 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     1561 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1621 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     1681 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     1741 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     1801 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     1861 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     1921 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     1981 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     2041 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     2101 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     2161 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     2221 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     2281 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     2341 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     2401 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     2461 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     2521 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     2581 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     2641 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     2701 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     2761 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     2821 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     2881 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     2941 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     3001 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     3061 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     3121 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     3181 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     3241 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     3301 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     3361 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     3421 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     3481 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     3541 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     3601 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     3661 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     3721 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     3781 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     3841 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3901 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     3961 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4021 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4081 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4141 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4201 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4261 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4321 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4381 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4441 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     4501 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4561 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4621 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4681 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4741 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     4801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4861 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     4921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5101 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     5161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     5221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5341 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     5401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5521 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     5581 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     5641 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     5701 atttaaatgg tcagttaga
//
