LOCUS Exported 4966 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4046627 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4966) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4966) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4966 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(258..1061) /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" primer_bind 1412..1430 /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" promoter complement(1734..2141) /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" terminator 2193..2224 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" promoter 2225..2327 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 2328..3188 /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 complement(2546..2565) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS 3189..3380 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" terminator 3404..3431 /label=T7Te terminator /note="phage T7 early transcription terminator" rep_origin complement(3443..4030) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4052..4081 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4159..175 /label=2u ori /note="yeast 2u plasmid origin of replication" ORIGIN 1 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata 61 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa 121 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttactcg 181 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt 241 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct 301 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt 361 ccaacaataa taatgtcaga tcctgtagaa accacatcat ccacggttct atactgttga 421 cccaatgcat ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa 481 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg 541 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat 601 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc 661 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat 721 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca 781 aattttctgt cctcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta 841 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt 901 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa 961 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta 1021 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg 1081 cacgtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca 1141 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt 1201 tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgtg tactgcagtg cactagtggt 1261 tcggcggccg ctttgtacag aaaaaaaaga aaaatttgaa atataaataa cgttcttaat 1321 actaacataa ctattaaaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt 1381 ccttttcggt tagagcggat gtgggaggag ggcgtgaatg taagcgtgac ataactaatt 1441 acatgatatc gacaaaggaa aaggggcctg tttataaggt tgccctgttg tccagcattt 1501 cttcacaaag cttgtttatg tctacagcac cctgcatgga aagcaaaaca gaaagtagtg 1561 aaaccatttt ttcaaaggct tcagtagtat ctttagctaa gagaatgtca ttgtgtaact 1621 ggacacattg agcccacaat ttagatgatg attctactct gagttgttgc aaaactgaga 1681 gtaagactac tgatgtgcac tttacatctg acattttcat tttgtaatta aaacttagat 1741 tagattgcta tgctttcttt ctaatgagca agaagtaaaa aaagttgtaa tagaacaaga 1801 aaaatgaaac tgaaacttga gaaattgaag accgtttatt aacttaaata tcaatgggag 1861 gtcatcgaaa gagaaaaaaa tcaaaaaaaa aaattttcaa gaaaaagaaa cgtgataaaa 1921 atttttattg cctttttcga cgaagaaaaa gaaacgaggc ggtctctttt ttcttttcca 1981 aacctttagt acgggtaatt aacgacaccc tagaggaaga aagaggggaa atttagtatg 2041 ctgtgcttgg gtgttttgaa gtggtacggc gatgcgcgga gtccgagaaa atctggaaga 2101 gtaaaaaagg agtagaaaca ttttgaagct atggtgtgtg gggcggccgc tctaactgac 2161 catttaaatc atacctgacc tccatagcag aaagtcaaaa gcctccgacc ggaggctttt 2221 gacttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg 2281 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg agtattcaac 2341 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 2401 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 2461 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt acgccccgaa gaacgttttc 2521 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 2581 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 2641 cagtcacaga aaagcatctc acggatggca tgacagtaag agaattatgc agtgctgcca 2701 taaccatgag tgataacact gcggccaact tacttctggc aacgatcgga ggaccgaagg 2761 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 2821 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 2881 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 2941 taatagactg gatggaggcg gataaagttg caggatcact tctgcgctcg gccctcccgg 3001 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 3061 cagcactggg gccagatggt aagccctccc gcatcgtagt tatctacacg acggggagtc 3121 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 3181 attggtaagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc 3241 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct 3301 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg 3361 atgacggtga aaacctctga tgagggccca aatgtaatca cctggctcac cttcgggtgg 3421 gcctttctgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 3481 atcgatgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgttta 3541 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 3601 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 3661 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 3721 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 3781 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 3841 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 3901 gcgctctgct gaagccagtt acctcggaaa aagagttggt agctcttgat ccggcaaaca 3961 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 4021 aggatctcaa gaagatcctt tgattttcta ccgaagaaag gcccacccgt gaaggtgagc 4081 cagtgagttg attgcagtcc agttacgctg gagtcaagct tgtaacacgc ggccgctcgg 4141 gaagttccta ttctctagct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa 4201 gcgctctgat gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa 4261 aatattgcga ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc 4321 tgtgctatat ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa 4381 ctcgacctct acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta 4441 gtaagaacta ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg 4501 tagtatatag agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat 4561 caacgctatc actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc 4621 ggggatgcct ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg 4681 gaagtggagt caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta 4741 accttaacgg acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag 4801 gagaaaaaaa gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt 4861 gtagaacaaa aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt 4921 ctgttctgta aaaatgcagc tcagattctt tgtttgaaaa attagc //