LOCUS Exported 4804 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4046353 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4804) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4804) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4804 /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 473..490 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 491..514 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(559..577) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 928..1731 /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 1814..2796 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 2874..2903 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 2925..3512 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 3524..3551 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3575..3766) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3767..4627) /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 4390..4409 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4628..4730) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 4731..4762 /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 tactcgtgcc 481 gagaggttct tggagccatc cgcagtttga aaaataaaca ggcccctttt cctttgtcga 541 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac 601 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt 661 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacaaagcg 721 gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta 781 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc 841 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca 901 aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct 961 actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac 1021 ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta 1081 ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag 1141 ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac 1201 agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga 1261 atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc 1321 ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca 1381 gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt 1441 gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga 1501 gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat 1561 gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt 1621 attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac 1681 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta 1741 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag 1801 ttattacccg agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag 1861 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc 1921 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac 1981 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc 2041 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat 2101 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt 2161 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag 2221 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt 2281 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat 2341 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt 2401 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata 2461 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat 2521 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga 2581 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata 2641 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta 2701 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg 2761 ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc 2821 ggccgcgtgt tacaagcttg actccagcgt aactggactg caatcaactc actggctcac 2881 cttcacgggt gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt 2941 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 3001 ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac 3061 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 3121 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 3181 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 3241 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 3301 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 3361 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg 3421 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt 3481 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc 3541 gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga 3601 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt 3661 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc 3721 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa 3781 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 3841 ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga 3901 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga 3961 gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt 4021 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 4081 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 4141 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 4201 gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 4261 cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt 4321 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 4381 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 4441 gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 4501 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 4561 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 4621 tactcatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta 4681 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc 4741 ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt 4801 taga //