LOCUS Exported 4320 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4133895 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4320) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4320) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4320 /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 71..136 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" polyA_signal 189..310 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(200..219) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 254..273 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 425..471 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 501..637 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 682..753 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(979..1567) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1059..1078) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2313..3173 /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(2531..2550) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3186..3322 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3474..3545 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3550..3596 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3614..3662 /note="synthetic polyadenylation signal" enhancer 3743..4046 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4048..4250 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4201..4221 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt 61 tcagtcgatg gattataagg accacgatgg agactacaaa gatcatgaca ttgattacaa 121 ggacgacgat gacaaataag ctagcttgac tgactgagat acagcgtacc ttcagctcac 181 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa 241 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa 301 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg 361 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt 421 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg 481 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg 541 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact 601 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg 661 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc 721 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga 781 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta 841 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat 901 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 961 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 1021 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 1081 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 1141 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 1201 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 1261 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 1321 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 1381 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 1441 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 1501 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 1561 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 1621 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 1681 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 1741 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 1801 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc 1861 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg 1921 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag 1981 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg 2041 aggaatatcg aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct 2101 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa 2161 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac 2221 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct 2281 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc 2341 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 2401 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca 2461 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt 2521 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg 2581 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc 2641 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 2701 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt 2761 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 2821 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta 2881 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg 2941 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 3001 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 3061 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 3121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc 3181 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca 3241 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc 3301 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga 3361 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg 3421 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg 3481 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac 3541 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa 3601 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta 3661 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc 3721 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac 3781 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa 3841 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 3901 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 3961 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 4021 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg 4081 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 4141 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 4201 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga 4261 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc //