LOCUS Exported 5133 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 2'-O-ribose methyltransferase. ACCESSION . VERSION . KEYWORDS pGBW-m4133218 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5133) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5133) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5133 /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..949 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 1002..1123 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1013..1032) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1067..1086 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1238..1284 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1314..1450 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1495..1566 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1792..2380) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1872..1891) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3126..3986 /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(3344..3363) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3999..4135 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4287..4358 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4363..4409 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4427..4475 /note="synthetic polyadenylation signal" enhancer 4556..4859 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4861..5063 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5014..5034 /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 cgtaccccta tgacgtacca gattatgcgt aagctagctt 961 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt 1021 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 1081 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat 1141 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct 1201 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct 1261 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg 1321 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc 1381 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat 1441 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc 1501 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc 1561 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta 1621 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg 1681 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc 1741 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 1801 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 1861 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 1921 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 1981 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 2041 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 2101 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 2161 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 2221 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 2281 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 2341 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 2401 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 2461 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 2521 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 2581 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag 2641 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa 2701 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat 2761 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac 2821 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg 2881 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt 2941 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag 3001 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa 3061 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct 3121 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 3181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 3241 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 3301 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 3361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 3421 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 3481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 3541 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 3601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 3661 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc 3721 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc 3781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 3841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 3901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 3961 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc 4021 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc 4081 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc 4141 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa 4201 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc 4261 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt 4321 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac 4381 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca 4441 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc 4501 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta 4561 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt 4621 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg 4681 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta 4741 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga 4801 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc 4861 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc 4921 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact 4981 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt 5041 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc 5101 taccatccac tcgacacacc cgccagcggc cgc //