LOCUS Exported 4272 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4133177 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4272) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4272) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4272 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 41..61 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 62..88 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 141..262 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(152..171) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 206..225 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 377..423 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 453..589 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 634..705 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(931..1519) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1011..1030) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2265..3125 /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(2483..2502) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3138..3274 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3426..3497 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3502..3548 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3566..3614 /note="synthetic polyadenylation signal" enhancer 3695..3998 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4000..4202 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4153..4173 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 4271..8 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc 61 gtacccctat gacgtaccag attatgcgta agctagcttg actgactgag atacagcgta 121 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat 181 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat 241 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca 301 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc 361 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt 421 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg 481 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc 541 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct 601 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca 661 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc 721 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct 781 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac 841 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag 901 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 961 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 1021 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 1081 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 1141 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 1201 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 1261 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 1321 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 1381 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 1441 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 1501 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 1561 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 1621 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 1681 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 1741 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca 1801 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg 1861 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc 1921 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct 1981 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct 2041 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt 2101 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg 2161 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt 2221 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt 2281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 2341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg 2401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa 2461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 2521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag 2581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 2641 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc 2701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 2761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 2821 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga 2881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 2941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 3001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 3061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 3121 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct 3181 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc 3241 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc 3301 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct 3361 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct 3421 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc 3481 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg 3541 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg 3601 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg 3661 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc 3721 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca 3781 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg 3841 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg 3901 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt 3961 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca 4021 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg 4081 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact 4141 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag 4201 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc 4261 gccagcggcc gc //