LOCUS Exported 4278 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4133395 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4278) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4278) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4278 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 47..67 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 68..94 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 147..268 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(158..177) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 212..231 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 383..429 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 459..595 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 640..711 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(937..1525) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1017..1036) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2271..3131 /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(2489..2508) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3144..3280 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3432..3503 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3508..3554 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3572..3620 /note="synthetic polyadenylation signal" enhancer 3701..4004 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4006..4208 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4159..4179 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt 61 tcagtcgtac ccctatgacg taccagatta tgcgtaagct agcttgactg actgagatac 121 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac 181 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt 241 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca 301 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat 361 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg 421 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc 481 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc 541 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg 601 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc 661 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg 721 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac 781 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta 841 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa 901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 1021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 1081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 1141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 1201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 1261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 1321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 1381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 1441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 1501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 1561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 1621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 1681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 1741 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa 1801 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt 1861 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc 1921 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc 1981 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga 2041 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc 2101 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc 2161 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc 2221 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc 2281 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 2341 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt 2401 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct 2461 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg 2521 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt 2581 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 2641 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga 2701 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 2761 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa 2821 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac 2881 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 2941 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 3001 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 3061 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 3121 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc 3181 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc 3241 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact 3301 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca 3361 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc 3421 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat 3481 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc 3541 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt 3601 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa 3661 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa 3721 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 3781 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 3841 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 3901 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 3961 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc 4021 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 4081 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 4141 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 4201 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac 4261 acacccgcca gcggccgc //