LOCUS       Exported                5391 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046559
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5391)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5391)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5391
                     /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             1018..1035
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1036..1101
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1146..1164)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1515..2318
                     /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      2401..3383
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3461..3490
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3512..4099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4111..4138
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4162..4353)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4354..5214)
                     /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     4977..4996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5215..5317)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5318..5349
                     /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 gaatctttgg tccctggatt taacgagaaa acccacgtac agctatccct tcccgtgttg
      541 caagttcgtg acgttctggt tcggggtttc ggtgattcag tcgaagaagt cttgagtgaa
      601 gccaggcaac atttaaagga tggtacttgt ggcttagttg aagtcgaaaa gggtgtgttg
      661 ccacaacttg agcaaccata cgttttcatt aagagatctg acgcaagaac agcgccacac
      721 ggtcatgtta tggttgaatt ggtcgctgaa ttggagggta tacaatatgg cagatcgggt
      781 gaaactttag gtgtattggt cccacacgta ggtgaaatcc ctgttgctta cagaaaagtt
      841 ttattaagaa agaatggtaa caagggtgct ggtggacact cctacggagc cgatctgaag
      901 tcttttgact tgggtgatga attgggtact gatccatatg aagacttcca agaaaattgg
      961 aacaccaaac atagctctgg tgttacgcgt gaattgatga gagaacttaa tggtggtctc
     1021 gtgccgagag gttctgatta taaagatcat gatggtgatt ataaagatca tgatattgat
     1081 tataaagatg atgatgataa ataaacaggc cccttttcct ttgtcgatat catgtaatta
     1141 gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga aaaggaagga
     1201 gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg ttagtattaa
     1261 gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc gccgaaccac
     1321 tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     1381 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     1441 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acgtgcagga
     1501 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     1561 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     1621 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     1681 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     1741 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga aaatttgctg
     1801 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     1861 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     1921 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     1981 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     2041 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     2101 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac
     2161 agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt gttggaagag
     2221 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     2281 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     2341 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccgagt
     2401 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     2461 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     2521 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     2581 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     2641 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     2701 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     2761 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     2821 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     2881 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     2941 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     3001 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     3061 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     3121 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     3181 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     3241 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     3301 cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc
     3361 tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc cgcgtgttac
     3421 aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt cacgggtggg
     3481 cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3541 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3601 taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa atactgttct
     3661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3901 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3961 cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct ggtatcttta
     4021 tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat gctcgtcagg
     4081 ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa ggtgagccag
     4141 gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     4201 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     4261 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     4321 catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca gtgaggcacc
     4381 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4441 aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4501 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag
     4561 aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4621 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4681 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4741 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4801 tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4861 tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc
     4921 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4981 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5041 taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5101 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5161 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc
     5221 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     5281 attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg
     5341 cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag a
//
