LOCUS Exported 7800 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast expression plasmid for SARS-CoV-2 nsp2. ACCESSION . VERSION . KEYWORDS pGBW-m4046172 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7800) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7800) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7800 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 25..466 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 25..142 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 437..460 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" CDS 2395..2412 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(3555..3573) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 3924..4727 /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 4810..5792 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 5870..5899 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 5921..6508 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 6520..6547 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(6571..6762) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(6763..7623) /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 7386..7405 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7624..7726) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 7727..7758 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg 61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg 121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat 181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta 241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag 301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt 361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta 421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg 481 gcatacacga ggtatgtaga caataacttt tgcggtcctg atggataccc actggaatgt 541 atcaaagatt tgttagctcg tgccggtaag gctagctgta ccctttctga gcaattggac 601 ttcatagaca ctaagagagg tgtttactgt tgtcgcgaac atgaacacga aattgcctgg 661 tataccgaga gatcagaaaa atcctacgaa ttacagactc cattcgaaat taagttggct 721 aagaagtttg atacattcaa cggcgaatgc cccaatttcg tctttccatt gaactctatc 781 attaaaacta tccaaccgag agtcgagaag aagaaattgg atggttttat gggtagaatt 841 agatcggtgt acccagttgc ttccccaaat gaatgtaacc aaatgtgtct atctactttg 901 atgaagtgtg accactgtgg tgaaaccagt tggcaaacag gcgacttcgt taaggctaca 961 tgtgaatttt gcggtaccga gaacctcact aaggaaggtg cgactacttg tggttattta 1021 cctcaaaatg cagttgtcaa gatttactgt ccagcttgcc ataactctga agttggacct 1081 gaacacagct tggctgaata ccacaacgaa tccggtttaa agaccatcct tcgaaaaggt 1141 ggtcgtacca tagccttcgg gggttgcgtc ttctcatatg ttggttgtca taataagtgt 1201 gcctattggg ttccaagagc ttctgctaat atcggatgta accacacggg tgtggtcggt 1261 gaaggctctg agggtttgaa cgataacttg ttggaaatac ttcagaagga aaaggtcaac 1321 attaacatcg ttggtgactt taaattgaat gaagaaattg ctattatcct agcgtccttc 1381 tcggcctcaa cttccgcttt tgtagaaact gttaagggtc ttgattacaa agctttcaag 1441 caaatcgtag aatcatgcgg taatttcaag gttaccaagg ggaaagccaa aaaaggtgcc 1501 tggaacattg gtgaacaaaa gtctatactg agtccattat atgctttcgc atctgaagct 1561 gctagagtcg tcagatctat tttttccaga actctcgaaa ccgctcaaaa ttcagtgaga 1621 gttttgcaaa aagcagccat tactatcttg gacggtattt cccagtactc tttgaggtta 1681 atcgatgcta tgatgtttac ctccgacttg gctactaata acctggtcgt catggcttac 1741 ataaccggtg gcgttgttca attgacatct caatggctaa ctaatatatt cggtactgtt 1801 tatgaaaagc tcaagccagt cttggattgg ttggaggaaa agttcaagga gggcgttgag 1861 tttttacggg atggttggga aatcgttaag ttcatttcta catgtgcctg tgaaattgtc 1921 ggtggtcaaa tcgtgacctg tgctaaggaa attaaggagt cggttcaaac tttctttaag 1981 ttggtgaaca aatttttagc tctctgtgct gattcgatta tcatcggagg agcgaaattg 2041 aaggcactga acttgggtga aaccttcgtc acccactcca aaggtctgta ccgtaaatgt 2101 gttaaatcaa gagaagaaac tggtttgcta atgcctttga aggccccgaa ggaaattatc 2161 ttcttggaag gtgaaacatt accaacagag gttttaacgg aagaggtagt tttgaagacg 2221 ggggatttgc agccgttaga acaaccaacc tctgaagccg tggaagctcc cctagttgga 2281 acgccagttt gcattaacgg tctgatgtta ttggaaatca aggacaccga aaagtactgt 2341 gctctcgctc caaacatgat ggtcactaac aataccttca ctttgaaggg tggtctcgtg 2401 ccgagaggtt ctaaaatcga ggaaggtaag ctagtcattt ggataaatgg agataagggt 2461 tacaacggtt tggcagaagt aggcaagaaa tttgaaaagg acacaggtat caaggtgacg 2521 gttgaacatc cagacaagtt agaagaaaaa ttccctcaag ttgctgctac tggtgatggt 2581 cccgacatta ttttctgggc ccacgatcgt tttggtggat atgctcagtc aggtttgctg 2641 gctgagatca ccccagacaa ggccttccaa gataaattgt acccattcac ttgggatgct 2701 gtcagataca acggtaagct tatagcatat ccaattgctg ttgaagcctt atcgttgatc 2761 tacaataagg acttgttacc taacccacca aagacatggg aagaaattcc agctttggat 2821 aaagagctaa aggccaaggg aaaaagcgct ttgatgttta acttacaaga accttatttc 2881 acctggccat taatcgctgc tgacggtggt tacgcattta aatacgaaaa cggtaagtat 2941 gacattaagg atgtcggcgt tgataatgct ggtgcaaagg ctggtttgac cttcttggtt 3001 gatttaataa agaataaaca catgaacgct gacactgact actctatcgc cgaagccgct 3061 ttcaacaagg gtgaaactgc tatgaccata aatggtccat gggcctggag taacattgat 3121 acttctaaag taaactatgg tgtcactgtt ttgcctacat tcaaaggcca accatccaag 3181 ccctttgtcg gtgttctatc cgctggcatc aatgctgctt ctccaaacaa ggagctagct 3241 aaagaatttt tggaaaatta cttgttaact gacgaaggtt tggaggccgt taacaaggac 3301 aagccattgg gtgctgtcgc tttgaaatct tacgaagaag aattggctaa ggatcccaga 3361 attgccgcta ccatggagaa cgcccaaaag ggtgaaatta tgccaaacat cccacaaatg 3421 tcagccttct ggtacgcagt taggactgcc gtgattaatg ctgcttccgg tagacaaacc 3481 gtcgacgaag cattaaaaga tgctcaaacg taaacaggcc ccttttcctt tgtcgatatc 3541 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa 3601 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt 3661 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg 3721 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct 3781 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat 3841 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa 3901 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc 3961 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt 4021 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc 4081 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca 4141 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa 4201 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag 4261 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt 4321 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat 4381 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga 4441 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg 4501 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat 4561 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg 4621 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa 4681 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat 4741 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat 4801 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg 4861 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa 4921 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga 4981 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg 5041 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat 5101 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc 5161 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg 5221 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac 5281 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc 5341 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc 5401 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa 5461 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt 5521 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca 5581 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata 5641 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc 5701 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt 5761 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc 5821 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc 5881 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc 5941 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 6001 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa 6061 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 6121 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 6181 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 6241 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 6301 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 6361 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg 6421 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg 6481 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag 6541 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg 6601 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc 6661 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat 6721 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag 6781 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 6841 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc 6901 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc 6961 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg 7021 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7081 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7141 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7201 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7261 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc 7321 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7381 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7441 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7501 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7561 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 7621 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga 7681 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc 7741 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga //