LOCUS Exported 4650 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp10. ACCESSION . VERSION . KEYWORDS pGBW-m4134050 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4650) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4650) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4650 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 81..384 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 386..588 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 539..559 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 657..666 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1077..1097 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1098..1124 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 1177..1298 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1188..1207) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1242..1261 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1413..1459 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1489..1625 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1670..1741 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1967..2555) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2047..2066) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3301..4161 /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(3519..3538) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4174..4310 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4462..4533 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4538..4584 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4602..4650 /note="synthetic polyadenylation signal" ORIGIN 1 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat 61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg 121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta 241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 361 gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga 421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac 601 cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgcca 661 ccatgggtaa tgcaacagaa gtgcctgcca attcaactgt attatctttc tgtgcttttg 721 ctgtagatgc tgctaaagct tacaaagatt atctagctag tgggggacaa ccaatcacta 781 attgtgttaa gatgttgtgt acacacactg gtactggtca ggcaataaca gttacaccgg 841 aagccaatat ggatcaagaa tcctttggtg gtgcatcgtg ttgtctgtac tgccgttgcc 901 acatagatca tccaaatcct aaaggatttt gtgacttaaa aggtaagtat gtacaaatac 961 ctacaacttg tgctaatgac cctgtgggtt ttacacttaa aaacacagtc tgtaccgtct 1021 gcggtatgtg gaaaggttat ggctgtagtt gtgatcaact ccgcgaaccc atgcttgaaa 1081 acctttactt tcagtcgtac ccctatgacg taccagatta tgcgtaagct agcttgactg 1141 actgagatac agcgtacctt cagctcacag acatgataag atacattgat gagtttggac 1201 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg 1261 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt 1321 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca 1381 aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg 1441 ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag 1501 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa 1561 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc 1621 cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa 1681 tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct 1741 aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg 1801 cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg 1861 gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag 1921 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 1981 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 2041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 2101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 2161 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 2221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 2281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 2341 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 2401 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 2461 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 2521 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 2581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 2641 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 2701 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 2761 cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct 2821 cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac 2881 tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca 2941 ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca 3001 tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta 3061 ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc 3121 agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg 3181 tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat 3241 aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc 3301 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 3361 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg 3421 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 3481 gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 3541 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 3601 gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 3661 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt 3721 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 3781 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 3841 cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct 3901 tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 3961 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 4021 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 4081 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 4141 tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat 4201 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc 4261 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat 4321 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc 4381 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt 4441 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc 4501 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt 4561 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca 4621 catccatctt cgatggatag cgattttatt //