LOCUS Exported 6376 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 leader protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046568 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6376) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6376) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6376 /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 971..988 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(2131..2149) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2500..3303 /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 3386..4368 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4446..4475 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4497..5084 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5096..5123 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(5147..5338) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5339..6199) /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 5962..5981 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6200..6302) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6303..6334 /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 atggaatctt tggtccctgg atttaacgag aaaacccacg tacagctatc 481 ccttcccgtg ttgcaagttc gtgacgttct ggttcggggt ttcggtgatt cagtcgaaga 541 agtcttgagt gaagccaggc aacatttaaa ggatggtact tgtggcttag ttgaagtcga 601 aaagggtgtg ttgccacaac ttgagcaacc atacgttttc attaagagat ctgacgcaag 661 aacagcgcca cacggtcatg ttatggttga attggtcgct gaattggagg gtatacaata 721 tggcagatcg ggtgaaactt taggtgtatt ggtcccacac gtaggtgaaa tccctgttgc 781 ttacagaaaa gttttattaa gaaagaatgg taacaagggt gctggtggac actcctacgg 841 agccgatctg aagtcttttg acttgggtga tgaattgggt actgatccat atgaagactt 901 ccaagaaaat tggaacacca aacatagctc tggtgttacg cgtgaattga tgagagaact 961 taatggtggt ctcgtgccga gaggttctaa aatcgaggaa ggtaagctag tcatttggat 1021 aaatggagat aagggttaca acggtttggc agaagtaggc aagaaatttg aaaaggacac 1081 aggtatcaag gtgacggttg aacatccaga caagttagaa gaaaaattcc ctcaagttgc 1141 tgctactggt gatggtcccg acattatttt ctgggcccac gatcgttttg gtggatatgc 1201 tcagtcaggt ttgctggctg agatcacccc agacaaggcc ttccaagata aattgtaccc 1261 attcacttgg gatgctgtca gatacaacgg taagcttata gcatatccaa ttgctgttga 1321 agccttatcg ttgatctaca ataaggactt gttacctaac ccaccaaaga catgggaaga 1381 aattccagct ttggataaag agctaaaggc caagggaaaa agcgctttga tgtttaactt 1441 acaagaacct tatttcacct ggccattaat cgctgctgac ggtggttacg catttaaata 1501 cgaaaacggt aagtatgaca ttaaggatgt cggcgttgat aatgctggtg caaaggctgg 1561 tttgaccttc ttggttgatt taataaagaa taaacacatg aacgctgaca ctgactactc 1621 tatcgccgaa gccgctttca acaagggtga aactgctatg accataaatg gtccatgggc 1681 ctggagtaac attgatactt ctaaagtaaa ctatggtgtc actgttttgc ctacattcaa 1741 aggccaacca tccaagccct ttgtcggtgt tctatccgct ggcatcaatg ctgcttctcc 1801 aaacaaggag ctagctaaag aatttttgga aaattacttg ttaactgacg aaggtttgga 1861 ggccgttaac aaggacaagc cattgggtgc tgtcgctttg aaatcttacg aagaagaatt 1921 ggctaaggat cccagaattg ccgctaccat ggagaacgcc caaaagggtg aaattatgcc 1981 aaacatccca caaatgtcag ccttctggta cgcagttagg actgccgtga ttaatgctgc 2041 ttccggtaga caaaccgtcg acgaagcatt aaaagatgct caaacgtaaa caggcccctt 2101 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac 2161 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt 2221 tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt 2281 ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt 2341 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg 2401 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa 2461 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag 2521 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa 2581 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta 2641 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat 2701 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta 2761 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg 2821 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca 2881 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt 2941 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt 3001 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac 3061 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat 3121 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga 3181 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag 3241 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac 3301 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt 3361 taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa 3421 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca 3481 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa 3541 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca 3601 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg 3661 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt 3721 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt 3781 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc 3841 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc 3901 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata 3961 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat 4021 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt 4081 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag 4141 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat 4201 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc 4261 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca 4321 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg 4381 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac 4441 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga 4501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 4561 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca 4621 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 4681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 4741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 4801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 4861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 4921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 4981 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 5041 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag 5101 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac 5161 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg 5221 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta 5281 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt 5341 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 5401 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca 5461 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 5521 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt 5581 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 5641 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 5701 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 5761 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca 5821 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg 5881 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 5941 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 6001 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca 6061 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 6121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 6181 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa 6241 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat 6301 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt 6361 taaatggtca gttaga //