LOCUS Exported 6199 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast expression plasmid for SARS-CoV-2 ORF7a protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046365 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6199) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6199) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6199 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(312..1115) /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" primer_bind 1466..1484 /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS complement(2627..2644) /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" promoter complement(3021..3428) /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" terminator 3480..3511 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" promoter 3512..3614 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 3615..4475 /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 complement(3833..3852) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS 4476..4667 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" terminator 4691..4718 /label=T7Te terminator /note="phage T7 early transcription terminator" rep_origin complement(4730..5317) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 5339..5368 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 5446..229 /label=2u ori /note="yeast 2u plasmid origin of replication" ORIGIN 1 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc 61 gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt 121 tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg 181 ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgtta ctcgggtaat 241 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat 301 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt 361 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca 421 ataataatgt cagatcctgt agaaaccaca tcatccacgg ttctatactg ttgacccaat 481 gcatctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca 541 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc 601 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat 661 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa 721 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct 781 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt 841 ctgtcctcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg 901 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga 961 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac 1021 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga 1081 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcacgtt 1141 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat 1201 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag 1261 attaccgaat caaaaaaatt tcaaggaaac cgtgtactgc agtgcactag tggttcggcg 1321 gccgctttgt acagaaaaaa aagaaaaatt tgaaatataa ataacgttct taatactaac 1381 ataactatta aaaaaaataa atagggacct agacttcagg ttgtctaact ccttcctttt 1441 cggttagagc ggatgtggga ggagggcgtg aatgtaagcg tgacataact aattacatga 1501 tatcgacaaa ggaaaagggg cctgtttacg tttgagcatc ttttaatgct tcgtcgacgg 1561 tttgtctacc ggaagcagca ttaatcacgg cagtcctaac tgcgtaccag aaggctgaca 1621 tttgtgggat gtttggcata atttcaccct tttgggcgtt ctccatggta gcggcaattc 1681 tgggatcctt agccaattct tcttcgtaag atttcaaagc gacagcaccc aatggcttgt 1741 ccttgttaac ggcctccaaa ccttcgtcag ttaacaagta attttccaaa aattctttag 1801 ctagctcctt gtttggagaa gcagcattga tgccagcgga tagaacaccg acaaagggct 1861 tggatggttg gcctttgaat gtaggcaaaa cagtgacacc atagtttact ttagaagtat 1921 caatgttact ccaggcccat ggaccattta tggtcatagc agtttcaccc ttgttgaaag 1981 cggcttcggc gatagagtag tcagtgtcag cgttcatgtg tttattcttt attaaatcaa 2041 ccaagaaggt caaaccagcc tttgcaccag cattatcaac gccgacatcc ttaatgtcat 2101 acttaccgtt ttcgtattta aatgcgtaac caccgtcagc agcgattaat ggccaggtga 2161 aataaggttc ttgtaagtta aacatcaaag cgctttttcc cttggccttt agctctttat 2221 ccaaagctgg aatttcttcc catgtctttg gtgggttagg taacaagtcc ttattgtaga 2281 tcaacgataa ggcttcaaca gcaattggat atgctataag cttaccgttg tatctgacag 2341 catcccaagt gaatgggtac aatttatctt ggaaggcctt gtctggggtg atctcagcca 2401 gcaaacctga ctgagcatat ccaccaaaac gatcgtgggc ccagaaaata atgtcgggac 2461 catcaccagt agcagcaact tgagggaatt tttcttctaa cttgtctgga tgttcaaccg 2521 tcaccttgat acctgtgtcc ttttcaaatt tcttgcctac ttctgccaaa ccgttgtaac 2581 ccttatctcc atttatccaa atgactagct taccttcctc gattttagaa cctctcggca 2641 cgagttcagt ctttcgcttc agtgtgaaac acaaggtaat gaaaacgata gcggcgacaa 2701 ttagaaaaat tggggagtac aactcttgaa cttcttcttg acggataaat aactttgggc 2761 ttacggatct cgctctcaat tgataaacgt gtttcacacc gtcgggacaa gcgaaagcga 2821 attgagtact aaaacaggtt aaggcgaact tgttatcagc caatggatga aatggcgaat 2881 tgccttcgta agttccagat gaacaaggtt ccttcaacaa gacagttgtc cccctaacgc 2941 attcctggta gtgataaagc tcacacgtag ccagggttat caatgctaag aataggataa 3001 ttttcatttt gtaattaaaa cttagattag attgctatgc tttctttcta atgagcaaga 3061 agtaaaaaaa gttgtaatag aacaagaaaa atgaaactga aacttgagaa attgaagacc 3121 gtttattaac ttaaatatca atgggaggtc atcgaaagag aaaaaaatca aaaaaaaaaa 3181 ttttcaagaa aaagaaacgt gataaaaatt tttattgcct ttttcgacga agaaaaagaa 3241 acgaggcggt ctcttttttc ttttccaaac ctttagtacg ggtaattaac gacaccctag 3301 aggaagaaag aggggaaatt tagtatgctg tgcttgggtg ttttgaagtg gtacggcgat 3361 gcgcggagtc cgagaaaatc tggaagagta aaaaaggagt agaaacattt tgaagctatg 3421 gtgtgtgggg cggccgctct aactgaccat ttaaatcata cctgacctcc atagcagaaa 3481 gtcaaaagcc tccgaccgga ggcttttgac ttgatcggca cgtaagaggt tccaactttc 3541 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag 3601 ctaaggaagc taaaatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 3661 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 3721 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 3781 agagtttacg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 3841 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 3901 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatctcacg gatggcatga 3961 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 4021 ttctggcaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 4081 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 4141 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 4201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 4261 gatcacttct gcgctcggcc ctcccggctg gctggtttat tgctgataaa tctggagccg 4321 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgca 4381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 4441 ctgagatagg tgcctcactg attaagcatt ggtaagtgac caaacaggaa aaaaccgccc 4501 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc 4561 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt 4621 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgatga gggcccaaat 4681 gtaatcacct ggctcacctt cgggtgggcc tttctgcgtt gctggcgttt ttccataggc 4741 tccgcccccc tgacgagcat cacaaaaatc gatgctcaag tcagaggtgg cgaaacccga 4801 caggactata aagataccag gcgtttaccc ctggaagctc cctcgtgcgc tctcctgttc 4861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 4981 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 5041 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 5101 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 5161 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc tcggaaaaag 5221 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 5281 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga ttttctaccg 5341 aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gcagtccagt tacgctggag 5401 tcaagcttgt aacacgcggc cgctcgggaa gttcctattc tctagctaga aagtatagga 5461 acttcagagc gcttttgaaa accaaaagcg ctctgatgac gcactttcaa aaaaccaaaa 5521 acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac aaacattgct 5581 caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct acccatccac 5641 ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt ttatctctag 5701 tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa tcgaaaacaa 5761 tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa gaaaccgttc 5821 ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac aaagtatgcg 5881 caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa atgcacccgc 5941 agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta tggaagagaa 6001 aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa aaagttatca 6061 agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg ctttgttaga 6121 aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga ttctttgttg 6181 gtaaaatagc gctctcgcg //