LOCUS Exported 4269 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4134024 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4269) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4269) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4269 /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..85 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 138..259 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(149..168) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 203..222 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 374..420 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 450..586 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 631..702 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(928..1516) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1008..1027) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2262..3122 /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(2480..2499) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3135..3271 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3423..3494 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3499..3545 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3563..3611 /note="synthetic polyadenylation signal" enhancer 3692..3995 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3997..4199 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4150..4170 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt 61 tcagtcgcac catcaccatc accattaagc tagcttgact gactgagata cagcgtacct 121 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca 181 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat 241 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg 301 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc 361 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg 421 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga 481 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac 541 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc 601 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca 661 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg 721 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg 781 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg 841 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa 901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 1021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 1081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 1141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 1201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 1261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 1321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 1381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 1441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 1501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 1561 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 1621 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 1681 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 1741 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac 1801 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc 1861 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga 1921 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc 1981 cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag 2041 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg 2101 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc 2161 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg 2221 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc 2281 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 2341 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac 2401 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga 2461 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 2521 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca 2581 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 2641 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa 2701 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 2761 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 2821 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag 2881 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 2941 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 3001 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 3061 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 3121 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat 3181 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata 3241 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg 3301 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt 3361 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc 3421 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc 3481 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta 3541 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata 3601 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt 3661 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 3721 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 3781 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 3841 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 3901 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 3961 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca 4021 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca 4081 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg 4141 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg 4201 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc 4261 agcggccgc //