LOCUS Exported 6510 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp2. ACCESSION . VERSION . KEYWORDS pGBW-m4133357 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6510) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6510) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6510 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 172..308 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 460..531 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 536..582 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 600..648 /note="synthetic polyadenylation signal" enhancer 729..1032 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1034..1236 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1187..1207 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 3222..3242 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 3591..3608 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 3685..3806 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3696..3715) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3750..3769 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 3921..3967 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 3997..4133 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4178..4249 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(4475..5063) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4555..4574) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 5809..159 /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(6027..6046) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc 61 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag 121 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg 181 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag 241 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt 301 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc 361 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta 421 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca 481 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa 541 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac 601 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg 661 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg 721 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 781 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 841 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 901 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 961 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 1021 gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 1081 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 1141 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 1201 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga 1261 tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgccacc atgtacactc 1321 gctatgtcga taacaacttc tgtggccctg atggctaccc tcttgagtgc attaaagacc 1381 ttctagcacg tgctggtaaa gcttcatgca ctttgtccga acaactggac tttattgaca 1441 ctaagagggg tgtatactgc tgccgtgaac atgagcatga aattgcttgg tacacggaac 1501 gttctgaaaa gagctatgaa ttgcagacac cttttgaaat taaattggca aagaaatttg 1561 acaccttcaa tggggaatgt ccaaattttg tatttccctt aaattccata atcaagacta 1621 ttcaaccaag ggttgaaaag aaaaagcttg atggctttat gggtagaatt cgatctgtct 1681 atccagttgc gtcaccaaat gaatgcaacc aaatgtgcct ttcaactctc atgaagtgtg 1741 atcattgtgg tgaaacttca tggcagacgg gcgattttgt taaagccact tgcgaatttt 1801 gtggcactga gaatttgact aaagaaggtg ccactacttg tggttactta ccccaaaatg 1861 ctgttgttaa aatttattgt ccagcatgtc acaattcaga agtaggacct gagcatagtc 1921 ttgccgaata ccataatgaa tctggcttga aaaccattct tcgtaagggt ggtcgcacta 1981 ttgcctttgg aggctgtgtg ttctcttatg ttggttgcca taacaagtgt gcctattggg 2041 ttccacgtgc tagcgctaac ataggttgta accatacagg tgttgttgga gaaggttccg 2101 aaggtcttaa tgacaacctt cttgaaatac tccaaaaaga gaaagtcaac atcaatattg 2161 ttggtgactt taaacttaat gaagagatcg ccattatttt ggcatctttt tctgcttcca 2221 caagtgcttt tgtggaaact gtgaaaggtt tggattataa agcattcaaa caaattgttg 2281 aatcctgtgg taattttaaa gttacaaaag gaaaagctaa aaaaggtgcc tggaatattg 2341 gtgaacagaa atcaatactg agtcctcttt atgcatttgc atcagaggct gctcgtgttg 2401 tacgatcaat tttctcccgc actcttgaaa ctgctcaaaa ttctgtgcgt gttttacaga 2461 aggccgctat aacaatacta gatggaattt cacagtattc actgagactc attgatgcta 2521 tgatgttcac atctgatttg gctactaaca atctagttgt aatggcctac attacaggtg 2581 gtgttgttca gttgacttcg cagtggctaa ctaacatctt tggcactgtt tatgaaaaac 2641 tcaaacccgt ccttgattgg cttgaagaga agtttaagga aggtgtagag tttcttagag 2701 acggttggga aattgttaaa tttatctcaa cctgtgcttg tgaaattgtc ggtggacaaa 2761 ttgtcacctg tgcaaaggaa attaaggaga gtgttcagac attctttaag cttgtaaata 2821 aatttttggc tttgtgtgct gactctatca ttattggtgg agctaaactt aaagccttga 2881 atttaggtga aacatttgtc acgcactcaa agggattgta cagaaagtgt gttaaatcca 2941 gagaagaaac tggcctactc atgcctctaa aagccccaaa agaaattatc ttcttagagg 3001 gagaaacact tcccacagaa gtgttaacag aggaagttgt cttgaaaact ggtgatttac 3061 aaccattaga acaacctact agtgaagctg ttgaagctcc attggttggt acaccagttt 3121 gtattaacgg gcttatgttg ctcgaaatca aagacacaga aaagtactgt gcccttgcac 3181 ctaatatgat ggtaacaaac aataccttca cactcaaagg cgaaaacctt tactttcagt 3241 cgatgagtga caaaatcatt catctaaccg atgactcctt tgatacagac gtgttgaagg 3301 ccgatggtgc gatactggta gatttctggg ctgaatggtg tggcccgtgc aagatgatcg 3361 cacccattct ggacgagatc gccgacgagt atcaaggaaa attaactgtc gctaagctta 3421 atatagatca gaacccaggg acggcaccta aatacggcat caggggaata cctaccctcc 3481 tgctctttaa gaatggggaa gttgccgcga caaaagtggg agccctgtct aagggccagc 3541 taaaagagtt cttggacgct aacctggcag gttcaggctc ggggcacatg caccatcacc 3601 atcaccatag cagcgggctc gtgccacgcg gctaagctag cttgactgac tgagatacag 3661 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta 3721 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa 3781 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg 3841 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg 3901 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg 3961 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct 4021 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt 4081 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc 4141 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg 4201 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt 4261 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt 4321 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt 4381 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg 4441 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4501 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4561 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4621 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4681 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 4741 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 4801 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 4861 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 4921 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 4981 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 5041 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 5101 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 5161 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 5221 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 5281 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc 5341 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct 5401 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca 5461 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg 5521 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc 5581 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca 5641 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt 5701 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta 5761 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa 5821 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 5881 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac 5941 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt 6001 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 6061 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca 6121 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 6181 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag 6241 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 6301 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 6361 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag 6421 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 6481 gctggctggt ttattgctga taaatctgga //