LOCUS Exported 6346 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 3'-to-5' exonuclease. ACCESSION . VERSION . KEYWORDS pGBW-m4046224 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6346) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6346) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6346 /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 2015..2032 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 2033..2056 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(2101..2119) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2470..3273 /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 3356..4338 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4416..4445 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4467..5054 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5066..5093 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5117..5308) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5309..6169) /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 5932..5951 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6170..6272) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6273..6304 /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 atggccgaga atgtcacagg cctattcaag gattgctcga aagtgatcac 481 cggtttacac cctacgcagg ctccaactca tttgtccgtt gacaccaagt ttaagactga 541 aggattgtgt gttgacatac caggtattcc caaagatatg acttaccgta ggctgatttc 601 tatgatgggt ttcaagatga actatcaagt taacggttac ccaaatatgt ttatcaccag 661 agaagaagct attagacacg tacgtgcatg gatcggtttc gatgtcgaag gttgtcatgc 721 tactagagaa gcggtcggta caaacttgcc attacaactt ggcttttcaa ccggtgttaa 781 tttggttgct gttcctacag ggtatgtcga cactccaaac aacacggatt tctctcgagt 841 gtccgccaag ccaccacctg gtgatcaatt caaacacttg atcccgctca tgtacaaggg 901 tttgccttgg aacgttgtca gaattaaaat agtacaaatg ttatctgata ctttgaagaa 961 tctgagtgac agagttgtct tcgtcctttg ggctcacggt tttgagctaa ccagcatgaa 1021 gtactttgtg aagattggac cagaacggac ttgttgcttg tgtgacagaa gagccacttg 1081 tttctccacc gctagcgata catatgcatg ttggcatcat tctattggtt tcgactacgt 1141 ttacaacccc tttatgatcg acgttcaaca gtggggtttc accggtaatt tgcaatcgaa 1201 ccacgacttg tattgtcaag ttcacggcaa tgctcacgtg gcttcatgcg atgctattat 1261 gacaaggtgt ttagccgtcc atgaatgttt cgttaaaaga gtagactgga ctatcgaata 1321 cccaattatc ggtgatgaac tcaagattaa cgctgcctgt agaaaggtcc aacatatggt 1381 tgtaaaggca gctctattag ctgacaaatt tccagttttg cacgatatag ggaatccaaa 1441 ggccataaag tgtgttccac aagctgacgt cgaatggaaa ttctacgatg cgcaaccctg 1501 ctctgataaa gcttataaga tcgaggaatt gttttactct tacgctaccc actccgataa 1561 gttcactgac ggtgtgtgtc ttttctggaa ctgcaatgtt gatagatatc cagcaaactc 1621 catcgtttgt agattcgaca cgcgagtctt gagcaacttg aacttaccag gttgtgatgg 1681 tggttccttg tacgtcaata aacatgcatt tcacactccc gctttcgaca agtctgcttt 1741 tgttaattta aagcaattgc cttttttcta ttactctgat agtccatgcg aatctcatgg 1801 taagcaagtc gtttccgaca ttgattacgt accactgaag tcagccacct gtataacaag 1861 atgtaacctt ggaggtgctg tctgccgcca ccatgctaac gagtacagac tatacttgga 1921 tgcctataac atgatgattt cagccggttt ctctttatgg gtttacaagc agtttgacac 1981 ttacaacttg tggaacacct tcacgcgttt gcaactcgtg ccgagaggtt cttggagcca 2041 tccgcagttt gaaaaataaa caggcccctt ttcctttgtc gatatcatgt aattagttat 2101 gtcacgctta cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag 2161 acaacctgaa gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg 2221 ttatttatat ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg 2281 cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa 2341 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa 2401 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga 2461 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt 2521 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat 2581 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta 2641 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta 2701 aaggcattat ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt 2761 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac 2821 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca 2881 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc 2941 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat 3001 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg 3061 ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat 3121 agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta 3181 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa 3241 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt 3301 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga 3361 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac 3421 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca 3481 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt 3541 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt 3601 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat 3661 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc 3721 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct 3781 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa 3841 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga 3901 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat 3961 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc 4021 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta 4081 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga 4141 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt 4201 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg 4261 cgtttttggt tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga 4321 agttcctata ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct 4381 tgactccagc gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt 4441 cttcggtaga aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 4501 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 4561 actctttttc cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 4621 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 4681 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 4741 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 4801 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 4861 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 4921 tcggaacagg agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc 4981 ctgtcgggtt tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc 5041 ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat 5101 tacatttggg ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc 5161 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt 5221 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt 5281 taagggcggt tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct 5341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 5401 cgatgcggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 5461 caccggctcc agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg 5521 atcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 5581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 5641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 5701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca 5761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 5821 ctgtcatgcc atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct 5881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 5941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac 6001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 6061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 6121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct 6181 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat 6241 ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt 6301 gactttctgc tatggaggtc aggtatgatt taaatggtca gttaga //