LOCUS Exported 5124 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 2'-O-ribose methyltransferase. ACCESSION . VERSION . KEYWORDS pGBW-m4133350 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5124) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5124) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5124 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 902..922 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 923..940 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 993..1114 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1004..1023) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1058..1077 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1229..1275 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1305..1441 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1486..1557 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1783..2371) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1863..1882) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3117..3977 /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(3335..3354) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3990..4126 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4278..4349 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4354..4400 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4418..4466 /note="synthetic polyadenylation signal" enhancer 4547..4850 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4852..5054 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5005..5025 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgtcc tcacaagcgt ggcagccagg cgtagcaatg cctaatttat acaagatgca 61 gcggatgctc cttgaaaaat gcgacctaca gaactatgga gatagtgcta cattgcccaa 121 ggggataatg atgaatgtgg ccaaatatac ccaactgtgt cagtacctga acactctcac 181 gctggccgtt ccgtacaaca tgagggtgat tcattttggt gccggcagcg acaagggagt 241 cgctcccggg accgctgtgc tgcgacagtg gcttcctaca ggcactctgc tagtggattc 301 tgatttgaat gacttcgtca gcgacgcaga ctcgaccctc atcggagatt gtgcgactgt 361 tcacacagcc aacaaatggg acctgatcat ttccgatatg tatgatccaa agaccaaaaa 421 tgtcacgaag gagaacgact caaaggaggg tttctttacg tacatctgcg ggttcatcca 481 acagaaactt gcactgggcg gaagtgtggc aattaaaata acagaacact cttggaacgc 541 cgacctatac aagcttatgg gtcattttgc ttggtggacc gcctttgtta ctaatgtaaa 601 cgcttcaagc tccgaggcct tcctgatagg ttgtaattat ttaggcaagc ctagagagca 661 gattgatggg tatgtgatgc acgcaaatta catcttctgg cgcaacacaa atccaattca 721 gctcagttct tattcactgt ttgatatgag caaattcccc cttaaactga gaggcactgc 781 cgtcatgtcc ttgaaggaag gacaaatcaa cgatatgatc ttgtctctcc ttagcaaggg 841 gcgtttgatt attagggaga acaatcgggt ggtgatctca tcggacgttc tggtcaacaa 901 tgaaaacctt tactttcagt cgcaccatca ccatcaccat taagctagct tgactgactg 961 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac 1021 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt 1081 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat 1141 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg 1201 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc 1261 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa 1321 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga 1381 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta 1441 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt 1501 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc 1561 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag 1621 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc 1681 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca 1741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 1801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 1861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 1921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 1981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 2041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 2101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 2161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 2221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 2281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 2341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 2401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 2461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 2521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 2581 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc 2641 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc 2701 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta 2761 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct 2821 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga 2881 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc 2941 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac 3001 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc 3061 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga 3121 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 3181 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag 3241 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 3301 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta 3361 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 3421 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 3481 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag 3541 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 3601 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 3661 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc 3721 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 3781 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 3841 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 3901 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 3961 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga 4021 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac 4081 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt 4141 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg 4201 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca 4261 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt 4321 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga 4381 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc 4441 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt 4501 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 4561 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 4621 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 4681 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 4741 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 4801 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt 4861 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc 4921 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat 4981 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 5041 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca 5101 ctcgacacac ccgccagcgg ccgc //