LOCUS Exported 5977 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 N (nucleocapsid). ACCESSION . VERSION . KEYWORDS pGBW-m4046669 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5977) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5977) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5977 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(1732..1750) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2101..2904 /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 2987..3969 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4047..4076 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4098..4685 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4697..4724 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4748..4939) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4940..5800) /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 5563..5582 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5801..5903) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5904..5935 /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 tttctnnnnc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga 301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta 361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt 421 taattacaaa atgtctgata atggacccca aaatcagcga aatgcacccc gcattacgtt 481 tggtggaccc tcagattcaa ctggcagtaa ccagaatgga gaacgcagtg gggcgcgatc 541 aaaacaacgt cggccccaag gtttacccaa taatactgcg tcttggttca ccgctctcac 601 tcaacatggc aaggaagacc ttaaattccc tcgaggacaa ggcgttccaa ttaacaccaa 661 tagcagtcca gatgaccaaa ttggctacta ccgaagagct accagacgaa ttcgtggtgg 721 tgacggtaaa atgaaagatc tcagtccaag atggtatttc tactacctag gaactgggcc 781 agaagctgga cttccctatg gtgctaacaa agacggcatc atatgggttg caactgaggg 841 agccttgaat acaccaaaag atcacattgg cacccgcaat cctgctaaca atgctgcaat 901 cgtgctacaa cttcctcaag gaacaacatt gccaaaaggc ttctacgcag aagggagcag 961 aggcggcagt caagcctctt ctcgttcctc atcacgtagt cgcaacagtt caagaaattc 1021 aactccaggc agcagtaggg gaacttctcc tgctagaatg gctggcaatg gcggtgatgc 1081 tgctcttgct ttgctgctgc ttgacagatt gaaccagctt gagagcaaaa tgtctggtaa 1141 aggccaacaa caacaaggcc aaactgtcac taagaaatct gctgctgagg cttctaagaa 1201 gcctcggcaa aaacgtactg ccactaaagc atacaatgta acacaagctt tcggcagacg 1261 tggtccagaa caaacccaag gaaattttgg ggaccaggaa ctaatcagac aaggaactga 1321 ttacaaacat tggccgcaaa ttgcacaatt tgcccccagc gcttcagcgt tcttcggaat 1381 gtcgcgcatt ggcatggaag tcacaccttc gggaacgtgg ttgacctaca caggtgccat 1441 caaattggat gacaaagatc caaatttcaa agatcaagtc attttgctga ataagcatat 1501 tgacgcatac aaaacattcc caccaacaga gcctaaaaag gacaaaaaga agaaggctga 1561 tgaaactcaa gccttaccgc agagacagaa gaaacagcaa actgtgactc ttcttcctgc 1621 tgcagatttg gatgatttct ccaaacaatt gcaacaatcc atgagcagtg ctgactcaac 1681 tcaggcctaa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt 1741 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga 1801 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata 1861 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt 1921 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg 1981 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat 2041 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc 2101 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag 2161 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc 2221 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca 2281 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta 2341 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca 2401 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat 2461 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta 2521 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct 2581 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc 2641 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg 2701 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg 2761 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag 2821 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg 2881 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac 2941 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt 3001 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct 3061 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat 3121 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag 3181 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa 3241 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac 3301 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa 3361 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct 3421 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt 3481 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata 3541 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg 3601 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt 3661 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag 3721 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat 3781 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat 3841 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg 3901 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat 3961 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag 4021 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag 4081 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4201 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 4261 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 4321 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 4381 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 4441 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 4501 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 4561 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt 4621 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat 4681 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg 4741 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct 4801 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt 4861 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg 4921 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct 4981 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg 5041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5101 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa 5161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 5221 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 5281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 5341 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt 5401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 5461 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 5521 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 5581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga 5641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 5701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 5761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg 5821 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt 5881 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg 5941 ctatggaggt caggtatgat ttaaatggtc agttaga //