LOCUS Exported 5002 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4046223 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5002) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5002) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5002 /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 677..694 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 695..712 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(757..775) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1126..1929 /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 2012..2994 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3072..3101 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3123..3710 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3722..3749 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3773..3964) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3965..4825) /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 4588..4607 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4826..4928) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 4929..4960 /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 atgaaaatgt cagatgtaaa gtgcacatca gtagtcttac tctcagtttt 481 gcaacaactc agagtagaat catcatctaa attgtgggct caatgtgtcc agttacacaa 541 tgacattctc ttagctaaag atactactga agcctttgaa aaaatggttt cactactttc 601 tgttttgctt tccatgcagg gtgctgtaga cataaacaag ctttgtgaag aaatgctgga 661 caacagggca accttactcg tgccgagagg ttctcaccat catcaccatc attaaacagg 721 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc 781 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat 841 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt 901 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga 961 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt 1021 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta 1081 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca 1141 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg 1201 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg 1261 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg 1321 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat 1381 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac 1441 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg 1501 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga 1561 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact 1621 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa 1681 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt 1741 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct 1801 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag 1861 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag 1921 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct 1981 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga 2041 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac 2101 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt 2161 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt 2221 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct 2281 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt 2341 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag 2401 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc 2461 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat 2521 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa 2581 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt 2641 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg 2701 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat 2761 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag 2821 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt 2881 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg 2941 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag 3001 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca 3061 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct 3121 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 3181 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct 3241 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 3301 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 3361 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 3421 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 3481 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 3541 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 3601 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 3661 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 3721 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt 3781 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt 3841 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa 3901 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg 3961 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 4021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg 4081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 4141 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca 4201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 4261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 4321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 4381 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat 4441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct 4501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 4561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 4621 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga 4681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 4741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 4801 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact 4861 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt 4921 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt 4981 atgatttaaa tggtcagtta ga //