LOCUS       Exported                6225 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046167
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6225)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6225)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6225
                     /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             820..837
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1980..1998)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2349..3152
                     /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      3235..4217
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4295..4324
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4346..4933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4945..4972
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4996..5187)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5188..6048)
                     /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     5811..5830
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6049..6151)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      6152..6183
                     /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 aacaatgagc tttctcctgt tgctttacgg cagatgtcct gcgcagctgg aacaacgcaa
      541 actgcctgta ctgacgataa cgctctagct tactataaca ccactaaagg tggtagattc
      601 gtcttggcct tgttgtcaga cctgcaagat ttgaagtggg cgagatttcc aaagagcgat
      661 ggcactggta ccatatacac cgaattagaa ccaccctgtc gtttcgtgac cgacactcca
      721 aaaggtccga aggttaagta tttgtacttt attaagggtt taaataactt gaatcgaggt
      781 atggttctgg ggtctttggc tgctacagtc agactccaac tcgtgccgag aggttctaaa
      841 atcgaggaag gtaagctagt catttggata aatggagata agggttacaa cggtttggca
      901 gaagtaggca agaaatttga aaaggacaca ggtatcaagg tgacggttga acatccagac
      961 aagttagaag aaaaattccc tcaagttgct gctactggtg atggtcccga cattattttc
     1021 tgggcccacg atcgttttgg tggatatgct cagtcaggtt tgctggctga gatcacccca
     1081 gacaaggcct tccaagataa attgtaccca ttcacttggg atgctgtcag atacaacggt
     1141 aagcttatag catatccaat tgctgttgaa gccttatcgt tgatctacaa taaggacttg
     1201 ttacctaacc caccaaagac atgggaagaa attccagctt tggataaaga gctaaaggcc
     1261 aagggaaaaa gcgctttgat gtttaactta caagaacctt atttcacctg gccattaatc
     1321 gctgctgacg gtggttacgc atttaaatac gaaaacggta agtatgacat taaggatgtc
     1381 ggcgttgata atgctggtgc aaaggctggt ttgaccttct tggttgattt aataaagaat
     1441 aaacacatga acgctgacac tgactactct atcgccgaag ccgctttcaa caagggtgaa
     1501 actgctatga ccataaatgg tccatgggcc tggagtaaca ttgatacttc taaagtaaac
     1561 tatggtgtca ctgttttgcc tacattcaaa ggccaaccat ccaagccctt tgtcggtgtt
     1621 ctatccgctg gcatcaatgc tgcttctcca aacaaggagc tagctaaaga atttttggaa
     1681 aattacttgt taactgacga aggtttggag gccgttaaca aggacaagcc attgggtgct
     1741 gtcgctttga aatcttacga agaagaattg gctaaggatc ccagaattgc cgctaccatg
     1801 gagaacgccc aaaagggtga aattatgcca aacatcccac aaatgtcagc cttctggtac
     1861 gcagttagga ctgccgtgat taatgctgct tccggtagac aaaccgtcga cgaagcatta
     1921 aaagatgctc aaacgtaaac aggccccttt tcctttgtcg atatcatgta attagttatg
     1981 tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga aggagttaga
     2041 caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta ttaagaacgt
     2101 tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa ccactagtgc
     2161 actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag
     2221 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
     2281 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa
     2341 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
     2401 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
     2461 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
     2521 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
     2581 aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg
     2641 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
     2701 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
     2761 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
     2821 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
     2881 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
     2941 tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt caacagtata
     3001 gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga agaggactat
     3061 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     3121 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta
     3181 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc gagtaacgaa
     3241 gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     3301 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa
     3361 cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc
     3421 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt
     3481 accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt
     3541 tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct
     3601 cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta
     3661 ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag
     3721 ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat
     3781 tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt
     3841 atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg
     3901 tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa
     3961 tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa
     4021 aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt
     4081 tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc
     4141 gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa
     4201 gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg ttacaagctt
     4261 gactccagcg taactggact gcaatcaact cactggctca ccttcacggg tgggcctttc
     4321 ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4381 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4441 ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4501 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4561 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4621 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4681 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4741 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4801 cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc tttatagtcc
     4861 tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt caggggggcg
     4921 gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt
     4981 acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     5041 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     5101 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     5161 aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg cacctatctc
     5221 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5281 gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5341 accggctcca gatttatcag caataaacca gccagccggg agggccgagc gcagaagtga
     5401 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5461 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5521 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5581 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgccag
     5641 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5701 tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5761 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5821 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact
     5881 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5941 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6001 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt tagcttcctt
     6061 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     6121 gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg
     6181 actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
