LOCUS Exported 6227 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4252808 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6227) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6227) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6227 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 26..74 /note="synthetic polyadenylation signal" terminator 92..138 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_feature 143..214 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 366..502 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" CDS complement(515..1375) /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 1138..1157 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" polyA_signal 1641..1689 /note="synthetic polyadenylation signal" CDS complement(1716..2315) /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1799..1819) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 2296..2315 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" promoter complement(2319..3528) /label=UbC promoter /note="human ubiquitin C promoter" primer_bind complement(2463..2482) /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" rep_origin 4028..4616 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4517..4536 /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 4842..4913 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4958..5094 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" terminator 5124..5170 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 5285..5406 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5322..5341) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5376..5395 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" CDS complement(5459..5524) /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS complement(5528..5548) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" promoter complement(5665..5867) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(5694..5714) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 5869..6172 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 ggctacactc gatcagcagt tagataataa aatcgctatc catcgaagat ggatgtgtgt 61 tggttttttg tgtgtgtaac gcaacgattg atagcataac cccttggggc ctctaaacgg 121 gtcttgaggg gttttttgaa ttgagaggta gctgaagctg ctgttatgac cactagaggg 181 aagaagatac ctgtggagct aatggtccaa gatgtcggga gccgcgatcg ctgccaaaaa 241 ggtaagctga cgatagctga cgagattcga tgcccgacga gcaaatgagc gccacggctg 301 ttcgcgtaag ctcgaaacga cggagattcg atagagccag agtcgagttt aaacgattcg 361 ggatcttaag gataaatttc tggtaaggag gacacgtatg gaagtgggca agttggggaa 421 gccgtatccg ttgctgaatc tggcatatgt gggagtataa gacgcgcagc gtcgcatcag 481 gcattttttt ctgcgccaat gcacctagaa tcggttacca atgcttaatc agtgaggcac 541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 601 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 661 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 721 gaagtggtcc tgcaacttta tccgcctcca tccagtctat caactgttgc cgggaagcta 781 gagtaagtag ttcgccagtt aatagtttac gcaaggttgt tgccattgct acaggcatcg 841 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 901 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccaatcg 961 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 1021 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac tcaaccaagt 1081 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 1141 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaagcgt tcttcggggc 1201 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgcgcac 1261 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 1321 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatgcgaa 1381 agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa agctaggact cgattatacg 1441 ttgaccgatc agactatcaa tatctaggtt taaacagatc ggtacgctgc cgtgaccagg 1501 cttgagtaca atatctgacg attggacttc ctgggtccca agctggattc cgactggctg 1561 actgccaagc aacgatatgg atcgtcgaga ctcgcactga gaggatcgtt ctcggtacac 1621 caggcgcgcc tagcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc 1681 gattttatta tctaagacta ggcttacata ctagcctagg caccgggctt gcgggtcatg 1741 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc 1801 tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc 1861 tcggcggcct ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg 1921 ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg 1981 aggccctcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg 2041 ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc 2101 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag 2161 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg 2221 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg 2281 gtggcgaggc gcaccgtggg cttgtactcg gtcatgttgt ctaacaaaaa agccaaaaac 2341 ggccagaatt tagcggacaa tttacaagtc taacactgaa aattacatat tgacccaaat 2401 gattacattt caaaaggtgc ctaaaaaact tcacaaaaca cactcgccaa ccccgagcgc 2461 atagttcaaa accggagctt cagctactta agaagatagg tacataaaac cgaccaaaga 2521 aactgacgcc tcacttatcc ctcccctcac cagaggtccg gcgcctgtcg attcaggaga 2581 gcctacccta ggcccgaacc ctgcgtcctg cgacggagaa aagcctaccg cacacctacc 2641 ggctggtggc cccaccctgc attataagcc aacagaacgg gtgacgtcac gacacgacga 2701 gggcgcgcgc tcccaaaggt acgggtgcac tgcccaacgg caccgccata actgccgccc 2761 ccgcaacaga cgacaaaccg agttctccag tcagtgacaa acttcacgtc agggtcccca 2821 gatggtgccc agcccatctc acccgaataa gagctttccc gcattagcga agggctcaag 2881 accttgggtt cttgccgccc accatgcccc ccaccttgtt tcaacgacct cacagcccgc 2941 ctcacaagcg tcttccattc aagactcggg aacagccgcc attttgctgc gctcccccca 3001 acccccagtt cagggcaacc ttgctcgcgg acccagacta cagcccttgg cggtctctcc 3061 acacgcttcc gtcccaccga gcggcccggc ggccacgaaa gccccggcca gcccagcagc 3121 ccgctactca ccaagtgacg atcacagcga tccacaaaca agaaccgcga cccaaatccc 3181 ggctgcgacg gaactagctg tgccacaccc ggcgcgtcct tatataatca tcggcgttca 3241 ccgccccacg gagatccctc cgcagaatcg ccgagaaggg actacttttc ctcgcctgtt 3301 ccgctctctg gaaagaaaac cagtgcccta gagtcaccca agtcccgtcc taaaatgtcc 3361 ttctgctgat actggggttc taaggccgag tcttatgagc agcgggccgc tgtcctgagc 3421 gtccgggcgg aaggatcagg acgctcctgc gcccttcgtc tgacgtggca gcgctcgccg 3481 tgacgagggg ggcgcccgcg ggaggcgcca aaacccggcg cggaggccag gcgcgccccg 3541 atttcgatat tcctcggatc tcggatagat cgggattgag gatgtaggtt gaacttctct 3601 cgggtgatcc cgatctcgga tattcgggat tggagtaata cctggcgata ggcgctgcca 3661 agcaatagcc aaggcgcaag gccggccaag agacttgcga tagttcgttc gagaggtagc 3721 ctgcaagatg gatcggacga aaggtttgaa gattttccga cgagggcctt ggagagttcg 3781 gaaatttaaa tgcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 3841 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa 3901 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 3961 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 4021 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 4081 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 4141 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 4201 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 4261 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 4321 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 4381 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 4441 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 4501 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 4561 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 4621 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgattt 4681 aaatgaaact cgtaatcgga gaacgggtgc aatagcaagg accagatcga acttgactga 4741 cgcaattctg agcgtagtcg aaacccagac aagtcgatct cgccaagtag ctgacttgac 4801 tggttaatta agtttcgata gcccaaggta accaatgaga ttagggaggg aagaaagttt 4861 gatgaactac ttctgacccc tagtggtgtc cagaaaagac cattaaagga atgtcttgat 4921 cgaaatcgag agatcgcctg caggatagct gacgacctta aggataaatt tctggtaagg 4981 aggacacgta tggaagtggg caagttgggg aagccgtatc cgttgctgaa tctggcatat 5041 gtgggagtat aagacgcgca gcgtcgcatc aggcattttt ttctgcgcca atgccggtag 5101 atagcaatcc ggcccgaggg gcacaaaaaa cccctcaaga cccgtttaga ggccccaagg 5161 ggttatgcta cgataccgat agagatgggc caataccaca tttgtagagg ttttacttgc 5221 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt 5281 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 5341 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 5401 atcttatcat gtctgtgagc tgaaggtacg ctgtatctca gtcagtcaag ctagcttatt 5461 tgtcatcgtc gtccttgtaa tcaatgtcat gatctttgta gtctccatcg tggtccttat 5521 aatccatcga ctgaaagtaa aggttttcca ctgcgaaacc gttgagaaag gactgagcat 5581 cggcactcat ggtggcggcc gctggcgggt gtgtcgagtg gatggtagga tcgacaaaga 5641 tctgatctga cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct 5701 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt 5761 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga 5821 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcagca tggtaatagc 5881 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc 5941 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg 6001 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca 6061 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg 6121 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc 6181 catatatggg ctatgaacta atgaccccgt aattgattac tagatct //