LOCUS Exported 5623 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp6. ACCESSION . VERSION . KEYWORDS pGBW-m4046474 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5623) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5623) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5623 /organism="synthetic DNA construct" /mol_type="other DNA" gap 385..395 /estimated_length=11 CDS 1298..1315 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 1316..1333 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1378..1396) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1747..2550 /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 2633..3510 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3693..3722 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3744..4331 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator 4343..4370 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(4394..4585) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4586..5446) /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 5209..5228 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5447..5549) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator 5550..5581 /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 tgctnnnnnn nnnnnaagca tagcaatcta atctaagttt 421 taattacaaa atggcagtga aaagaacaat caagggtaca caccactggt tgttactcac 481 aattttgact tcacttttag ttttagtcca gagtactcaa tggtctttgt tctttttttt 541 gtatgaaaat gcctttttac cttttgctat gggtattatt gctatgtctg cttttgcaat 601 gatgtttgtc aaacataagc atgcatttct ctgtttgttt ttgttacctt ctcttgccac 661 tgtagcttat tttaatatgg tctatatgcc tgctagttgg gtgatgcgta ttatgacatg 721 gttggatatg gttgatacta gtttgtctgg ttttaagcta aaagactgtg ttatgtatgc 781 atcagctgta gtgttactaa tccttatgac agcaagaact gtgtatgatg atggtgctag 841 gagagtgtgg acacttatga atgtcttgac actcgtttat aaagtttatt atggtaatgc 901 tttagatcaa gccatttcca tgtgggctct tataatctct gttacttcta actactcagg 961 tgtagttaca actgtcatgt ttttggccag aggtattgtt tttatgtgtg ttgagtattg 1021 ccctattttc ttcataactg gtaatacact tcagtgtata atgctagttt attgtttctt 1081 aggctatttt tgtacttgtt actttggcct cttttgttta ctcaaccgct actttagact 1141 gactcttggt gtttatgatt acttagtttc tacacaggag tttagatata tgaattcaca 1201 gggactactc ccacccaaga atagcataga tgccttcaaa ctcaacatta aattgttggg 1261 tgttggtggc aaaccttgta tcaaagtagc cactgtactc gtgccgagag gttctcacca 1321 tcatcaccat cattaaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc 1381 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca 1441 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta 1501 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac 1561 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa 1621 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca 1681 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga 1741 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct 1801 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt 1861 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta 1921 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag 1981 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt 2041 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt 2101 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa 2161 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc 2221 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt 2281 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg 2341 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga 2401 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt 2461 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca 2521 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata 2581 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc 2641 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa 2701 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg 2761 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa 2821 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac 2881 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt 2941 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct 3001 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt 3061 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct 3121 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg 3181 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat 3241 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta 3301 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata 3361 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag 3421 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg 3481 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt 3541 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt 3601 tcctatactn nnnagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga 3661 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt 3721 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 3781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 3841 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 3901 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 3961 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 4021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 4081 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 4141 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 4201 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg 4261 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga 4321 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac 4381 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt 4441 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca 4501 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa 4561 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag 4621 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 4681 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 4741 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc 4801 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 4861 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 4921 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 4981 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa 5041 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 5101 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 5161 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 5221 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct 5281 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 5341 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 5401 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag 5461 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt 5521 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac 5581 tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga //