LOCUS Exported 5755 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 endoRNAse. ACCESSION . VERSION . KEYWORDS pGBW-m4046625 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5755) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5755) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5755 /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(1510..1528) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1879..2682 /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 2765..3747 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3825..3854 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3876..4463 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4475..4502 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4526..4717) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4718..5578) /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 5341..5360 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5579..5681) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5682..5713 /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 atgttagaaa atgtggcttt taatgttgta aataagggac actttgatgg 481 acaacagggt gaagtaccag tttctatcat taataacact gtttacacaa aagttgatgg 541 tgttgatgta gaattgtttg aaaataaaac aacattacct gttaatgtag catttgagct 601 ttgggctaag cgcaacatta aaccagtacc agaggtgaaa atactcaata atttgggtgt 661 ggacattgct gctaatactg tgatctggga ctacaaaaga gatgctccag cacatatatc 721 tactattggt gtttgttcta tgactgacat agccaagaaa ccaactgaaa cgatttgtgc 781 accactcact gtcttttttg atggtagagt tgatggtcaa gtagacttat ttagaaatgc 841 ccgtaatggt gttcttatta cagaaggtag tgttaaaggt ttacaaccat ctgtaggtcc 901 caaacaagct agtcttaatg gagtcacatt aattggagaa gccgtaaaaa cacagttcaa 961 ttattataag aaagttgatg gtgttgtcca acaattacct gaaacttact ttactcagag 1021 tagaaattta caagaattta aacccaggag tcaaatggaa attgatttct tagaattagc 1081 tatggatgaa ttcattgaac ggtataaatt agaaggctat gccttcgaac atatcgttta 1141 tggagatttt agtcatagtc agttaggtgg tttacatcta ctgattggac tagctaaacg 1201 ttttaaggaa tcaccttttg aattagaaga ttttattcct atggacagta cagttaaaaa 1261 ctatttcata acagatgcgc aaacaggttc atctaagtgt gtgtgttctg ttattgattt 1321 attacttgat gattttgttg aaataataaa atcccaagat ttatctgtag tttctaaggt 1381 tgtcaaagtg actattgact atacagaaat ttcatttatg ctttggtgta aagatggcca 1441 tgtagaaaca ttttacccaa aattataaac aggccccttt tcctttgtcg atatcatgta 1501 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga 1561 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta 1621 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa 1681 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa 1741 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt 1801 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc 1861 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct 1921 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct 1981 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa 2041 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt 2101 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt 2161 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa 2221 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag 2281 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca 2341 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc 2401 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt 2461 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt 2521 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga 2581 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca 2641 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta 2701 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc 2761 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat 2821 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct 2881 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg 2941 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga 3001 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga 3061 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat 3121 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact 3181 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt 3241 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac 3301 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg 3361 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt 3421 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc 3481 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc 3541 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa 3601 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac 3661 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa 3721 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg 3781 ttacaagctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg 3841 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta 3901 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 3961 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg 4021 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 4081 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 4141 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 4201 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 4261 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 4321 gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc 4381 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt 4441 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag 4501 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga 4561 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt 4621 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc 4681 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg 4741 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 4801 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 4861 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc 4921 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 4981 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5041 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 5101 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5161 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5221 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca 5281 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5341 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5401 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5461 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5521 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt 5581 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta 5641 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg 5701 gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag ttaga //