LOCUS Exported 6673 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp2. ACCESSION . VERSION . KEYWORDS pGBW-m4046450 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6673) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6673) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6673 /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 2348..2365 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 2366..2383 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2428..2446) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2797..3600 /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 3683..4665 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4743..4772 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4794..5381 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5393..5420 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5444..5635) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5636..6496) /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 6259..6278 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6497..6599) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6600..6631 /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 atggcataca cgaggtatgt agacaataac ttttgcggtc ctgatggata 481 cccactggaa tgtatcaaag atttgttagc tcgtgccggt aaggctagct gtaccctttc 541 tgagcaattg gacttcatag acactaagag aggtgtttac tgttgtcgcg aacatgaaca 601 cgaaattgcc tggtataccg agagatcaga aaaatcctac gaattacaga ctccattcga 661 aattaagttg gctaagaagt ttgatacatt caacggcgaa tgccccaatt tcgtctttcc 721 attgaactct atcattaaaa ctatccaacc gagagtcgag aagaagaaat tggatggttt 781 tatgggtaga attagatcgg tgtacccagt tgcttcccca aatgaatgta accaaatgtg 841 tctatctact ttgatgaagt gtgaccactg tggtgaaacc agttggcaaa caggcgactt 901 cgttaaggct acatgtgaat tttgcggtac cgagaacctc actaaggaag gtgcgactac 961 ttgtggttat ttacctcaaa atgcagttgt caagatttac tgtccagctt gccataactc 1021 tgaagttgga cctgaacaca gcttggctga ataccacaac gaatccggtt taaagaccat 1081 ccttcgaaaa ggtggtcgta ccatagcctt cgggggttgc gtcttctcat atgttggttg 1141 tcataataag tgtgcctatt gggttccaag agcttctgct aatatcggat gtaaccacac 1201 gggtgtggtc ggtgaaggct ctgagggttt gaacgataac ttgttggaaa tacttcagaa 1261 ggaaaaggtc aacattaaca tcgttggtga ctttaaattg aatgaagaaa ttgctattat 1321 cctagcgtcc ttctcggcct caacttccgc ttttgtagaa actgttaagg gtcttgatta 1381 caaagctttc aagcaaatcg tagaatcatg cggtaatttc aaggttacca aggggaaagc 1441 caaaaaaggt gcctggaaca ttggtgaaca aaagtctata ctgagtccat tatatgcttt 1501 cgcatctgaa gctgctagag tcgtcagatc tattttttcc agaactctcg aaaccgctca 1561 aaattcagtg agagttttgc aaaaagcagc cattactatc ttggacggta tttcccagta 1621 ctctttgagg ttaatcgatg ctatgatgtt tacctccgac ttggctacta ataacctggt 1681 cgtcatggct tacataaccg gtggcgttgt tcaattgaca tctcaatggc taactaatat 1741 attcggtact gtttatgaaa agctcaagcc agtcttggat tggttggagg aaaagttcaa 1801 ggagggcgtt gagtttttac gggatggttg ggaaatcgtt aagttcattt ctacatgtgc 1861 ctgtgaaatt gtcggtggtc aaatcgtgac ctgtgctaag gaaattaagg agtcggttca 1921 aactttcttt aagttggtga acaaattttt agctctctgt gctgattcga ttatcatcgg 1981 aggagcgaaa ttgaaggcac tgaacttggg tgaaaccttc gtcacccact ccaaaggtct 2041 gtaccgtaaa tgtgttaaat caagagaaga aactggtttg ctaatgcctt tgaaggcccc 2101 gaaggaaatt atcttcttgg aaggtgaaac attaccaaca gaggttttaa cggaagaggt 2161 agttttgaag acgggggatt tgcagccgtt agaacaacca acctctgaag ccgtggaagc 2221 tcccctagtt ggaacgccag tttgcattaa cggtctgatg ttattggaaa tcaaggacac 2281 cgaaaagtac tgtgctctcg ctccaaacat gatggtcact aacaatacct tcactttgaa 2341 gggtggtctc gtgccgagag gttctcacca tcatcaccat cattaaacag gccccttttc 2401 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc 2461 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt 2521 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg 2581 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt 2641 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta 2701 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg 2761 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa 2821 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag 2881 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt 2941 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt 3001 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc 3061 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt 3121 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt 3181 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg 3241 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact 3301 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg 3361 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac 3421 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct 3481 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt 3541 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa 3601 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa 3661 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg 3721 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa 3781 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa 3841 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa 3901 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc 3961 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt 4021 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa 4081 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac 4141 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc 4201 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg 4261 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata 4321 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct 4381 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc 4441 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc 4501 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat 4561 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag 4621 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac 4681 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca 4741 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat 4801 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 4861 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag 4921 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 4981 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 5041 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 5101 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 5161 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 5221 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 5281 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc 5341 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa 5401 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt 5461 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt 5521 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg 5581 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc 5641 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 5701 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg 5761 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 5821 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta 5881 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 5941 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 6001 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 6061 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg 6121 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga 6181 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 6241 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 6301 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt 6361 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 6421 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 6481 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata 6541 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa 6601 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa 6661 atggtcagtt aga //