LOCUS       Exported                5406 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046183
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5406)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5406
                     /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             1075..1092
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1093..1116
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1161..1179)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1530..2333
                     /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      2416..3398
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3476..3505
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3527..4114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4126..4153
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4177..4368)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4369..5229)
                     /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     4992..5011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5230..5332)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5333..5364
                     /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 gctatcgcca gtgaattttc ctcattacct agctatgcag ctttcgctac ggctcaggag
      541 gcctacgaac aagctgtagc caacggcgac tctgaagttg ttcttaagaa attgaagaag
      601 tctttgaatg tcgcgaaatc tgaatttgat agagatgctg ctatgcaacg taagctagaa
      661 aagatggctg accaagcaat gacccaaatg tacaaacaag ccagaagtga ggacaagcga
      721 gctaaggtga cttccgctat gcaaacaatg ctgttcacta tgttgagaaa gttggataac
      781 gatgctctga acaatattat aaacaatgcc agagatggtt gtgtccccct caacattatc
      841 ccattaacca ctgccgctaa attgatggtt gttattccgg attataatac atacaaaaac
      901 acttgcgacg gtaccacctt cacatacgcc tctgctcttt gggaaatcca acaggtagtc
      961 gacgctgact ccaagatagt tcaattgagc gaaatttcta tggataactc gccaaaccta
     1021 gcttggccat taatcgtcac tgcgttgaga gctaatagtg cggttaagtt gcaactcgtg
     1081 ccgagaggtt cttggagcca tccgcagttt gaaaaataaa caggcccctt ttcctttgtc
     1141 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     1201 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     1261 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     1321 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     1381 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     1441 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     1501 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     1561 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     1621 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     1681 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     1741 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     1801 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     1861 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     1921 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     1981 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     2041 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     2101 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     2161 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     2221 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     2281 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     2341 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     2401 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     2461 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2521 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     2581 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     2641 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     2701 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     2761 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     2821 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     2881 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     2941 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     3001 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3061 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     3121 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     3181 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     3241 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     3301 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     3361 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     3421 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     3481 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     3541 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3601 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     3661 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3721 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3781 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3841 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3901 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3961 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     4021 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     4081 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     4141 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     4201 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4261 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4321 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     4381 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4441 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     4501 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4561 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     4621 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4681 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4741 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4801 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4861 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     4921 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4981 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5041 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5101 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5161 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5221 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     5281 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     5341 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     5401 gttaga
//
