LOCUS       Exported                5223 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046290
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5223)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5223)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5223
                     /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             892..909
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             910..933
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(978..996)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1347..2150
                     /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      2233..3215
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3293..3322
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3344..3931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3943..3970
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3994..4185)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4186..5046)
                     /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     4809..4828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5047..5149)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5150..5181
                     /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 ggtaatgcaa cagaagtgcc tgccaattca actgtattat ctttctgtgc ttttgctgta
      541 gatgctgcta aagcttacaa agattatcta gctagtgggg gacaaccaat cactaattgt
      601 gttaagatgt tgtgtacaca cactggtact ggtcaggcaa taacagttac accggaagcc
      661 aatatggatc aagaatcctt tggtggtgca tcgtgttgtc tgtactgccg ttgccacata
      721 gatcatccaa atcctaaagg attttgtgac ttaaaaggta agtatgtaca aatacctaca
      781 acttgtgcta atgaccctgt gggttttaca cttaaaaaca cagtctgtac cgtctgcggt
      841 atgtggaaag gttatggctg tagttgtgat caactccgcg aacccatgct tctcgtgccg
      901 agaggttctt ggagccatcc gcagtttgaa aaataaacag gccccttttc ctttgtcgat
      961 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
     1021 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
     1081 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg
     1141 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa
     1201 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
     1261 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     1321 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     1381 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     1441 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     1501 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     1561 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca
     1621 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     1681 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     1741 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     1801 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     1861 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     1921 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg
     1981 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta
     2041 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     2101 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     2161 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     2221 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     2281 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     2341 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     2401 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     2461 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     2521 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     2581 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     2641 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     2701 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     2761 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     2821 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     2881 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     2941 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     3001 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     3061 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     3121 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt
     3181 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg
     3241 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc
     3301 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     3361 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3421 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     3481 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3541 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3601 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3661 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3721 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3781 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc
     3841 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     3901 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     3961 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa
     4021 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     4081 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     4141 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat
     4201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4261 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat
     4321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag
     4381 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4621 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4681 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta
     4741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4861 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5041 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     5101 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc
     5161 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt
     5221 aga
//
