LOCUS Exported 4329 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4133333 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4329) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4329) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4329 /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..145 /codon_start=1 /product="calmodulin-binding peptide" /label=CBP /note="derived from skeletal muscle myosin light chain kinase; binds calmodulin with nanomolar affinity in the presence of calcium" /translation="KRRWKKNFIAVSAANRFKKISSSGAL" polyA_signal 198..319 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(209..228) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 263..282 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 434..480 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 510..646 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 691..762 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(988..1576) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1068..1087) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2322..3182 /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(2540..2559) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3195..3331 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3483..3554 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3559..3605 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3623..3671 /note="synthetic polyadenylation signal" enhancer 3752..4055 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4057..4259 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4210..4230 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt 61 tcagtcgaag aggcgatgga aaaagaattt tatcgcggtg agtgcagcca acagattcaa 121 aaagatatcc tcgtctggtg ccctctaagc tagcttgact gactgagata cagcgtacct 181 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca 241 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat 301 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg 361 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc 421 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg 481 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga 541 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac 601 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc 661 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca 721 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg 781 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg 841 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg 901 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa 961 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 1021 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 1081 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 1141 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 1201 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 1261 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 1321 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 1381 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 1441 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 1501 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 1561 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 1621 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 1681 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 1741 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 1801 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac 1861 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc 1921 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga 1981 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc 2041 cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag 2101 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg 2161 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc 2221 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg 2281 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc 2341 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 2401 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac 2461 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga 2521 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 2581 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca 2641 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 2701 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa 2761 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 2821 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 2881 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag 2941 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 3001 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 3061 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 3121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 3181 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat 3241 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata 3301 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg 3361 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt 3421 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc 3481 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc 3541 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta 3601 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata 3661 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt 3721 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 3781 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 3841 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 3901 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 3961 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 4021 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca 4081 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca 4141 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg 4201 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg 4261 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc 4321 agcggccgc //