LOCUS Exported 5098 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp9. ACCESSION . VERSION . KEYWORDS pGBW-m4046272 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5098) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5098) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5098 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 10..417 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" CDS 773..790 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 791..808 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(853..871) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1222..2025 /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 2108..3090 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3168..3197 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3219..3806 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3818..3845 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3869..4060) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4061..4921) /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 4684..4703 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4922..5024) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5025..5056 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat 61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat 121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa 181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg 241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga 301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta 361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt 421 taattacaaa atgaacaatg agctttctcc tgttgcttta cggcagatgt cctgcgcagc 481 tggaacaacg caaactgcct gtactgacga taacgctcta gcttactata acaccactaa 541 aggtggtaga ttcgtcttgg ccttgttgtc agacctgcaa gatttgaagt gggcgagatt 601 tccaaagagc gatggcactg gtaccatata caccgaatta gaaccaccct gtcgtttcgt 661 gaccgacact ccaaaaggtc cgaaggttaa gtatttgtac tttattaagg gtttaaataa 721 cttgaatcga ggtatggttc tggggtcttt ggctgctaca gtcagactcc aactcgtgcc 781 gagaggttct caccatcatc accatcatta aacaggcccc ttttcctttg tcgatatcat 841 gtaattagtt atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa 901 ggaaggagtt agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta 961 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc 1021 gaaccactag tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc 1081 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt 1141 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg 1201 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat 1261 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt 1321 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc 1381 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca 1441 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa 1501 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca 1561 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg 1621 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg 1681 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag 1741 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa 1801 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg 1861 ggtcaacagt atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt 1921 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa 1981 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa 2041 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta 2101 cccgagtaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct 2161 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc 2221 gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga 2281 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg 2341 agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc 2401 cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc 2461 tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct 2521 actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg 2581 attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta 2641 taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat 2701 tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat 2761 gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt 2821 gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag 2881 ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc 2941 aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt 3001 tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca 3061 aaagcgctct gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc 3121 gtgttacaag cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac 3181 gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc 3241 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 3301 caagagctac caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata 3361 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 3421 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 3481 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 3541 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 3601 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 3661 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt 3721 atctttatag tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct 3781 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt 3841 gagccaggtg attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg 3901 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct 3961 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa 4021 agcgggccat gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg 4081 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 4141 tgtagataac tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc 4201 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg 4261 agcgcagaag tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg 4321 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 4381 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 4441 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 4501 cgatcgttgc cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 4561 ataattctct tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa 4621 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 4681 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 4741 cggggcgtaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 4801 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 4861 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 4921 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc 4981 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg 5041 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga //