LOCUS Exported 4762 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4046673 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4762) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4762) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4762 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 10..417 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" primer_bind complement(517..535) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 886..1689 /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 1772..2754 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 2832..2861 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 2883..3470 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3482..3509 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3533..3724) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3725..4585) /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 4348..4367 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4586..4688) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 4689..4720 /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 atgtccgctg acgcacaatc attcctaaat gggtttgctg tataaacagg 481 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc 541 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat 601 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt 661 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga 721 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt 781 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta 841 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca 901 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg 961 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg 1021 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg 1081 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat 1141 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac 1201 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg 1261 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga 1321 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact 1381 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa 1441 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt 1501 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct 1561 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag 1621 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag 1681 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct 1741 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga 1801 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac 1861 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt 1921 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt 1981 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct 2041 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt 2101 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag 2161 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc 2221 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat 2281 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa 2341 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt 2401 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg 2461 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat 2521 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag 2581 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt 2641 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg 2701 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag 2761 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca 2821 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct 2881 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 2941 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct 3001 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 3061 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 3121 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 3181 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 3241 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 3301 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 3361 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 3421 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 3481 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt 3541 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt 3601 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa 3661 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg 3721 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 3781 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg 3841 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 3901 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca 3961 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 4021 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 4081 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 4141 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat 4201 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct 4261 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 4321 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 4381 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga 4441 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 4501 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 4561 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact 4621 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt 4681 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt 4741 atgatttaaa tggtcagtta ga //