LOCUS Exported 4224 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4136048 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4224) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4224) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4224 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 93..214 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(104..123) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 158..177 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 329..375 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 405..541 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 586..657 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(883..1471) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(963..982) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2217..3077 /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(2435..2454) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3090..3226 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3378..3449 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3454..3500 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3518..3566 /note="synthetic polyadenylation signal" enhancer 3647..3950 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3952..4154 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4105..4125 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 4223..8 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg taagctagct tgactgactg 61 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac 121 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt 181 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat 241 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg 301 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc 361 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa 421 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga 481 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta 541 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt 601 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc 661 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag 721 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc 781 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca 841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 1021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 1081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 1141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 1201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 1261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 1321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 1381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 1441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 1501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 1561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 1621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 1681 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc 1741 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc 1801 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta 1861 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct 1921 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga 1981 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc 2041 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac 2101 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc 2161 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga 2221 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 2281 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag 2341 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 2401 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta 2461 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 2521 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 2581 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag 2641 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 2701 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 2761 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc 2821 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 2881 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 2941 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 3001 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 3061 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga 3121 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac 3181 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt 3241 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg 3301 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca 3361 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt 3421 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga 3481 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc 3541 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt 3601 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 3661 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 3721 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 3781 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 3841 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 3901 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt 3961 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc 4021 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat 4081 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 4141 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca 4201 ctcgacacac ccgccagcgg ccgc //