LOCUS       Exported                6651 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046343
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6651)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6651)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6651
                     /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             2278..2295
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2296..2361
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2406..2424)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2775..3578
                     /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      3661..4643
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4721..4750
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4772..5359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      5371..5398
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5422..5613)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5614..6474)
                     /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     6237..6256
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6475..6577)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      6578..6609
                     /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 gttggggctt gtgttctttg caattcacag acttcattaa gatgtggtgc ttgcatacgt
      541 agaccattct tatgttgtaa atgctgttac gaccatgtca tatcaacatc acataaatta
      601 gtcttgtctg ttaatccgta tgtttgcaat gctccaggtt gtgatgtcac agatgtgact
      661 caactttact taggaggtat gagctattat tgtaaatcac ataaaccacc cattagtttt
      721 ccattgtgtg ctaatggaca agtttttggt ttatataaaa atacatgtgt tggtagcgat
      781 aatgttactg actttaatgc aattgcaaca tgtgactgga caaatgctgg tgattacatt
      841 ttagctaaca cctgtactga aagactcaag ctttttgcag cagaaacgct caaagctact
      901 gaggagacat ttaaactgtc ttatggtatt gctactgtac gtgaagtgct gtctgacaga
      961 gaattacatc tttcatggga agttggtaaa cctagaccac cacttaaccg aaattatgtc
     1021 tttactggtt atcgtgtaac taaaaacagt aaagtacaaa taggagagta cacctttgaa
     1081 aaaggtgact atggtgatgc tgttgtttac cgaggtacaa caacttacaa attaaatgtt
     1141 ggtgattatt ttgtgctgac atcacataca gtaatgccat taagtgcacc tacactagtg
     1201 ccacaagagc actatgttag aattactggc ttatacccaa cactcaatat ctcagatgag
     1261 ttttctagca atgttgcaaa ttatcaaaag gttggtatgc aaaagtattc tacactccag
     1321 ggaccacctg gtactggtaa gagtcatttt gctattggcc tagctctcta ctacccttct
     1381 gctcgcatag tgtatacagc ttgctctcat gccgctgttg atgcactatg tgagaaggca
     1441 ttaaaatatt tgcctataga taaatgtagt agaattatac ctgcacgtgc tcgtgtagag
     1501 tgttttgata aattcaaagt gaattcaaca ttagaacagt atgtcttttg tactgtaaat
     1561 gcattgcctg agacgacagc agatatagtt gtctttgatg aaatttcaat ggccacaaat
     1621 tatgatttga gtgttgtcaa tgccagatta cgtgctaagc actatgtgta cattggcgac
     1681 cctgctcaat tacctgcacc acgcacattg ctaactaagg gcacactaga accagaatat
     1741 ttcaattcag tgtgtagact tatgaaaact ataggtccag acatgttcct cggaacttgt
     1801 cggcgttgtc ctgctgaaat tgttgacact gtgagtgctt tggtttatga taataagctt
     1861 aaagcacata aagacaaatc agctcaatgc tttaaaatgt tttataaggg tgttatcacg
     1921 catgatgttt catctgcaat taacaggcca caaataggcg tggtaagaga attccttaca
     1981 cgtaaccctg cttggagaaa agctgtcttt atttcacctt ataattcaca gaatgctgta
     2041 gcctcaaaga ttttgggact accaactcaa actgttgatt catcacaggg ctcagaatat
     2101 gactatgtca tattcactca aaccactgaa acagctcact cttgtaatgt aaacagattt
     2161 aatgttgcta ttaccagagc aaaagtaggc atactttgca taatgtctga tagagacctt
     2221 tatgacaagt tgcaatttac aagtcttgaa attccacgta ggaatgtggc aactttactc
     2281 gtgccgagag gttctgatta taaagatcat gatggtgatt ataaagatca tgatattgat
     2341 tataaagatg atgatgataa ataaacaggc cccttttcct ttgtcgatat catgtaatta
     2401 gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga aaaggaagga
     2461 gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg ttagtattaa
     2521 gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc gccgaaccac
     2581 tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     2641 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     2701 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acgtgcagga
     2761 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     2821 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     2881 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     2941 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     3001 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga aaatttgctg
     3061 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     3121 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     3181 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     3241 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     3301 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     3361 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac
     3421 agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt gttggaagag
     3481 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     3541 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     3601 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccgagt
     3661 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     3721 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     3781 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     3841 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     3901 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     3961 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     4021 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     4081 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     4141 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     4201 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     4261 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     4321 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     4381 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     4441 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     4501 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     4561 cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc
     4621 tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc cgcgtgttac
     4681 aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt cacgggtggg
     4741 cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4801 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4861 taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa atactgttct
     4921 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4981 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5041 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5101 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5161 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5221 cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct ggtatcttta
     5281 tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat gctcgtcagg
     5341 ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa ggtgagccag
     5401 gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     5461 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     5521 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     5581 catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca gtgaggcacc
     5641 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5701 aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5761 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag
     5821 aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5881 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5941 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6001 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6061 tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6121 tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc
     6181 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6241 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6301 taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6361 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6421 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc
     6481 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     6541 attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg
     6601 cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag a
//
