LOCUS Exported 6137 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4252974 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6137) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6137) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6137 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 250..321 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 326..372 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 390..438 /note="synthetic polyadenylation signal" enhancer 519..822 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 824..1026 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 977..997 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" polyA_signal 1195..1316 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1206..1225) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1260..1279 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1431..1477 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1507..1643 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1688..1759 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1985..2573) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2065..2084) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 3073..4282 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 4119..4138 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 4286..4885 /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(4286..4305) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4782..4802 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 4912..4960 /note="synthetic polyadenylation signal" CDS 5226..6086 /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(5444..5463) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 6099..98 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" ORIGIN 1 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac 61 ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg 121 actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt 181 tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc 241 ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa 301 cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa 361 ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg 421 atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt 481 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat 541 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt 601 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa 661 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc 721 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct 781 acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag 841 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt 901 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac 961 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc 1021 agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac 1081 acccgccagc ggccgccacc atgagtgccg atgctcagtc ctttctcaac ggtttcgcag 1141 tgtaagctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat 1201 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg 1261 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca 1321 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa 1381 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc 1441 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat 1501 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac 1561 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct 1621 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg 1681 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc 1741 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca 1801 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg 1861 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt 1921 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 1981 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 2041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 2101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 2161 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 2221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 2281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 2341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 2401 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 2461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 2521 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 2581 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 2641 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 2701 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 2761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc 2821 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag 2881 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct 2941 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg 3001 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat 3061 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca 3121 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac 3181 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa 3241 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg 3301 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg 3361 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc 3421 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg 3481 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt 3541 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt 3601 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag 3661 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt 3721 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc 3781 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg 3841 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc 3901 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc 3961 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc 4021 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt 4081 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat 4141 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc 4201 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc 4261 gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac 4321 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc 4381 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact 4441 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc 4501 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg 4561 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct 4621 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc 4681 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga 4741 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga 4801 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg 4861 catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc 4921 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg 4981 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt 5041 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag 5101 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa 5161 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct 5221 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 5281 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 5341 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 5401 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 5461 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 5521 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 5581 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 5641 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 5701 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 5761 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc 5821 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc 5881 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 5941 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 6001 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 6061 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc 6121 ctgatgcgac gctgcgc //