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" promoter 10..417 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" CDS 794..811 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(1954..1972) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 2323..3126 /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 3209..4191 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 4269..4298 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 4320..4907 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4919..4946 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4970..5161) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5162..6022) /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 5785..5804 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6023..6125) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 6126..6157 /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 atgaaaatta tcctattctt agcattgata accctggcta cgtgtgagct 481 ttatcactac caggaatgcg ttagggggac aactgtcttg ttgaaggaac cttgttcatc 541 tggaacttac gaaggcaatt cgccatttca tccattggct gataacaagt tcgccttaac 601 ctgttttagt actcaattcg ctttcgcttg tcccgacggt gtgaaacacg tttatcaatt 661 gagagcgaga tccgtaagcc caaagttatt tatccgtcaa gaagaagttc aagagttgta 721 ctccccaatt tttctaattg tcgccgctat cgttttcatt accttgtgtt tcacactgaa 781 gcgaaagact gaactcgtgc cgagaggttc taaaatcgag gaaggtaagc tagtcatttg 841 gataaatgga gataagggtt acaacggttt ggcagaagta ggcaagaaat ttgaaaagga 901 cacaggtatc aaggtgacgg ttgaacatcc agacaagtta gaagaaaaat tccctcaagt 961 tgctgctact ggtgatggtc ccgacattat tttctgggcc cacgatcgtt ttggtggata 1021 tgctcagtca ggtttgctgg ctgagatcac cccagacaag gccttccaag ataaattgta 1081 cccattcact tgggatgctg tcagatacaa cggtaagctt atagcatatc caattgctgt 1141 tgaagcctta tcgttgatct acaataagga cttgttacct aacccaccaa agacatggga 1201 agaaattcca gctttggata aagagctaaa ggccaaggga aaaagcgctt tgatgtttaa 1261 cttacaagaa ccttatttca cctggccatt aatcgctgct gacggtggtt acgcatttaa 1321 atacgaaaac ggtaagtatg acattaagga tgtcggcgtt gataatgctg gtgcaaaggc 1381 tggtttgacc ttcttggttg atttaataaa gaataaacac atgaacgctg acactgacta 1441 ctctatcgcc gaagccgctt tcaacaaggg tgaaactgct atgaccataa atggtccatg 1501 ggcctggagt aacattgata cttctaaagt aaactatggt gtcactgttt tgcctacatt 1561 caaaggccaa ccatccaagc cctttgtcgg tgttctatcc gctggcatca atgctgcttc 1621 tccaaacaag gagctagcta aagaattttt ggaaaattac ttgttaactg acgaaggttt 1681 ggaggccgtt aacaaggaca agccattggg tgctgtcgct ttgaaatctt acgaagaaga 1741 attggctaag gatcccagaa ttgccgctac catggagaac gcccaaaagg gtgaaattat 1801 gccaaacatc ccacaaatgt cagccttctg gtacgcagtt aggactgccg tgattaatgc 1861 tgcttccggt agacaaaccg tcgacgaagc attaaaagat gctcaaacgt aaacaggccc 1921 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc 1981 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta 2041 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt 2101 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat 2161 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga 2221 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc 2281 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat 2341 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac 2401 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag 2461 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact 2521 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt 2581 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct 2641 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc 2701 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc 2761 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag 2821 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga 2881 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta 2941 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca 3001 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta 3061 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa 3121 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca 3181 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca 3241 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga 3301 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta 3361 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt 3421 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt 3481 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact 3541 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc 3601 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga 3661 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa 3721 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg 3781 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc 3841 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat 3901 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta 3961 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga 4021 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt 4081 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca 4141 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat 4201 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc 4261 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct 4321 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 4381 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca 4441 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 4501 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 4561 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 4621 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 4681 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 4741 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 4801 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 4861 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 4921 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt 4981 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa 5041 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg 5101 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca 5161 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 5221 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc 5281 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 5341 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc 5401 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 5461 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 5521 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 5581 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac 5641 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt 5701 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 5761 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 5821 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat 5881 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 5941 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 6001 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa 6061 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc 6121 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg 6181 atttaaatgg tcagttaga //