LOCUS Exported 5296 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 leader protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046308 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5296) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5296) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5296 /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" CDS 989..1006 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1051..1069) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1420..2223 /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 2306..3288 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3366..3395 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3417..4004 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4016..4043 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4067..4258) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4259..5119) /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 4882..4901 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5120..5222) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5223..5254 /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 gaggttctca ccatcatcac catcattaaa caggcccctt 1021 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac 1081 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt 1141 tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt 1201 ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt 1261 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg 1321 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa 1381 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag 1441 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa 1501 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta 1561 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat 1621 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta 1681 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg 1741 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca 1801 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt 1861 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt 1921 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac 1981 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat 2041 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga 2101 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag 2161 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac 2221 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt 2281 taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa 2341 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca 2401 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa 2461 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca 2521 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg 2581 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt 2641 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt 2701 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc 2761 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc 2821 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata 2881 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat 2941 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt 3001 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag 3061 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat 3121 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc 3181 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca 3241 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg 3301 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac 3361 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga 3421 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 3481 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca 3541 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 3601 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 3661 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 3721 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 3781 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 3841 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 3901 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 3961 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag 4021 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac 4081 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg 4141 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta 4201 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt 4261 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 4321 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca 4381 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 4441 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt 4501 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 4561 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 4621 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 4681 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca 4741 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg 4801 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 4861 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 4921 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca 4981 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 5041 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 5101 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa 5161 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat 5221 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt 5281 taaatggtca gttaga //