LOCUS Exported 5094 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp6. ACCESSION . VERSION . KEYWORDS pGBW-m4134108 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5094) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5094) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5094 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 872..892 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 893..910 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 963..1084 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(974..993) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1028..1047 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1199..1245 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1275..1411 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1456..1527 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1753..2341) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1833..1852) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3087..3947 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3305..3324) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3960..4096 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4248..4319 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4324..4370 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4388..4436 /note="synthetic polyadenylation signal" enhancer 4517..4820 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4822..5024 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4975..4995 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 5093..8 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 caccatggca gtgaaaagaa caatcaaggg tacacaccac tggttgttac tcacaatttt 61 gacttcactt ttagttttag tccagagtac tcaatggtct ttgttctttt ttttgtatga 121 aaatgccttt ttaccttttg ctatgggtat tattgctatg tctgcttttg caatgatgtt 181 tgtcaaacat aagcatgcat ttctctgttt gtttttgtta ccttctcttg ccactgtagc 241 ttattttaat atggtctata tgcctgctag ttgggtgatg cgtattatga catggttgga 301 tatggttgat actagtttgt ctggttttaa gctaaaagac tgtgttatgt atgcatcagc 361 tgtagtgtta ctaatcctta tgacagcaag aactgtgtat gatgatggtg ctaggagagt 421 gtggacactt atgaatgtct tgacactcgt ttataaagtt tattatggta atgctttaga 481 tcaagccatt tccatgtggg ctcttataat ctctgttact tctaactact caggtgtagt 541 tacaactgtc atgtttttgg ccagaggtat tgtttttatg tgtgttgagt attgccctat 601 tttcttcata actggtaata cacttcagtg tataatgcta gtttattgtt tcttaggcta 661 tttttgtact tgttactttg gcctcttttg tttactcaac cgctacttta gactgactct 721 tggtgtttat gattacttag tttctacaca ggagtttaga tatatgaatt cacagggact 781 actcccaccc aagaatagca tagatgcctt caaactcaac attaaattgt tgggtgttgg 841 tggcaaacct tgtatcaaag tagccactgt agaaaacctt tactttcagt cgcaccatca 901 ccatcaccat taagctagct tgactgactg agatacagcg taccttcagc tcacagacat 961 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt 1021 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca 1081 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt 1141 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta 1201 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg 1261 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat 1321 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata 1381 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct 1441 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt 1501 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta 1561 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca 1621 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag 1681 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 1741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 1801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 1861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 1921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 1981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 2041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 2101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 2161 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 2221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 2281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 2341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 2401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 2461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 2521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt 2581 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca 2641 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg 2701 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg 2761 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat 2821 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga 2881 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa 2941 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat 3001 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag 3061 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc 3121 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 3181 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg 3241 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag 3301 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 3361 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta 3421 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 3481 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca 3541 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 3601 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat 3661 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg 3721 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 3781 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 3841 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 3901 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg 3961 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc 4021 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc 4081 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc 4141 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc 4201 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat 4261 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca 4321 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg 4381 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct 4441 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc 4501 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 4561 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 4621 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 4681 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 4741 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 4801 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc 4861 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 4921 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 4981 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc 5041 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc //