LOCUS       Exported                5265 bp ds-DNA     circular SYN 16-JUN-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046580
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5265)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5265)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 16, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5265
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             868..885
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             886..951
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(996..1014)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1365..2168
                     /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      2251..3233
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3311..3340
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3362..3949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3961..3988
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4012..4203)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4204..5064)
                     /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     4827..4846
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5065..5167)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5168..5199
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 cggattagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatggaaa agctgcataa ccactttaac taatactttc aacattttca
      361 gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaact ataatgggta atgcaacaga agtgcctgcc
      481 aattcaactg tattatcttt ctgtgctttt gctgtagatg ctgctaaagc ttacaaagat
      541 tatctagcta gtgggggaca accaatcact aattgtgtta agatgttgtg tacacacact
      601 ggtactggtc aggcaataac agttacaccg gaagccaata tggatcaaga atcctttggt
      661 ggtgcatcgt gttgtctgta ctgccgttgc cacatagatc atccaaatcc taaaggattt
      721 tgtgacttaa aaggtaagta tgtacaaata cctacaactt gtgctaatga ccctgtgggt
      781 tttacactta aaaacacagt ctgtaccgtc tgcggtatgt ggaaaggtta tggctgtagt
      841 tgtgatcaac tccgcgaacc catgcttctc gtgccgagag gttctgatta taaagatcat
      901 gatggtgatt ataaagatca tgatattgat tataaagatg atgatgataa ataaacaggc
      961 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct
     1021 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1081 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     1141 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa
     1201 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     1261 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     1321 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat
     1381 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     1441 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     1501 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     1561 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     1621 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     1681 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     1741 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     1801 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     1861 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     1921 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     1981 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta
     2041 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     2101 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     2161 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     2221 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa
     2281 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2341 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     2401 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     2461 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     2521 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     2581 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     2641 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     2701 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     2761 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     2821 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     2881 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     2941 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     3001 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     3061 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     3121 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     3181 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga
     3241 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa
     3301 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt
     3361 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3421 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt
     3481 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3541 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3601 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3661 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3721 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3781 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3841 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3901 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3961 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt
     4021 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     4081 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     4141 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     4201 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4261 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg
     4321 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4381 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat
     4441 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4501 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4561 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4621 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc
     4681 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt
     4741 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4801 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4861 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag
     4921 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4981 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5041 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc
     5101 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     5161 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     5221 tgatttaaat ggtcagttag agcggccgct tgctatattg aagta
//
