LOCUS Exported 4263 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4133855 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4263) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4263) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4263 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 26..35 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 68..88 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 89..106 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 159..280 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(170..189) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 224..243 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 395..441 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 471..607 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 652..723 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(949..1537) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1029..1048) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2283..3143 /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(2501..2520) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3156..3292 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3444..3515 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3520..3566 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3584..3632 /note="synthetic polyadenylation signal" enhancer 3713..4016 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4018..4220 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4171..4191 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 ccactcgaca cacccgccag cggccgccac catggctgat gcacaatcgt ttttaaacgg 61 gtttgcggaa aacctttact ttcagtcgca ccatcaccat caccattaag ctagcttgac 121 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg 181 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat 241 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca 301 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta 361 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa 421 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc 481 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc 541 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta 601 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt 661 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc 721 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt 781 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc 841 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa 901 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 961 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 1021 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 1081 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 1141 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 1201 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 1261 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 1321 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 1381 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 1441 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 1501 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 1561 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 1621 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 1681 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 1741 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc 1801 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga 1861 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc 1921 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta 1981 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg 2041 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag 2101 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct 2161 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt 2221 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc 2281 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 2341 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 2401 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 2461 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 2521 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 2581 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 2641 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 2701 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 2761 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 2821 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag 2881 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc 2941 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 3001 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 3061 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 3121 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg 3181 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc 3241 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga 3301 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag 3361 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt 3421 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc 3481 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg 3541 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca 3601 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga 3661 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 3721 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 3781 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 3841 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 3901 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 3961 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga 4021 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa 4081 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc 4141 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg 4201 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac 4261 cat //