LOCUS Exported 5655 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 2'-O-ribose methyltransferase. ACCESSION . VERSION . KEYWORDS pGBW-m4133339 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5655) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5655) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5655 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 896..916 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 950..1027 /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" CDS 1055..1075 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" polyA_signal 1524..1645 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1535..1554) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1589..1608 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1760..1806 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1836..1972 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 2017..2088 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(2314..2902) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2394..2413) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3648..4508 /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(3866..3885) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4521..4657 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4809..4880 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4885..4931 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4949..4997 /note="synthetic polyadenylation signal" enhancer 5078..5381 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5383..5585 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5536..5556 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagt caagcgtggc aaccgggtgt tgctatgcct aatctttaca aaatgcaaag 61 aatgctatta gaaaagtgtg accttcaaaa ttatggtgat agtgcaacat tacctaaagg 121 cataatgatg aatgtcgcaa aatatactca actgtgtcaa tatttaaaca cattaacatt 181 agctgtaccc tataatatga gagttataca ttttggtgct ggttctgata aaggagttgc 241 accaggtaca gctgttttaa gacagtggtt gcctacgggt acgctgcttg tcgattcaga 301 tcttaatgac tttgtctctg atgcagattc aactttgatt ggtgattgtg caactgtaca 361 tacagctaat aaatgggatc tcattattag tgatatgtac gaccctaaga ctaaaaatgt 421 tacaaaagaa aatgactcta aagagggttt tttcacttac atttgtgggt ttatacaaca 481 aaagctagct cttggaggtt ccgtggctat aaagataaca gaacattctt ggaatgctga 541 tctttataag ctcatgggac acttcgcatg gtggacagcc tttgttacta atgtgaatgc 601 gtcatcatct gaagcatttt taattggatg taattatctt ggcaaaccac gcgaacaaat 661 agatggttat gtcatgcatg caaattacat attttggagg aatacaaatc caattcagtt 721 gtcttcctat tctttatttg acatgagtaa atttcccctt aaattaaggg gtactgctgt 781 tatgtcttta aaagaaggtc aaatcaatga tatgatttta tctcttctta gtaaaggtag 841 acttataatt agagaaaaca acagagttgt tatttctagt gatgttcttg ttaacgaaaa 901 cctttacttt cagtcgggac ggcgaatccc ggggctgata aatccctgga aaaggagatg 961 gaagaagaac tttattgcag tgtctgcggc taaccggttc aaaaagatca gttcaagcgg 1021 cgccctagat tacgacattc ctacgaccgc ctccgagaat ctctatttcc agggtgaatt 1081 tggcttggca caacatgacg aggccgtcga taacaagttt aataaagaac agcagaacgc 1141 tttctatgag atccttcact taccaaatct gaacgaagag caacgcaacg ctttcattca 1201 gagcctgaaa gacgatccct cgcagtctgc aaatctcctg gccgaagcca agaagctgaa 1261 tgacgcacag gctccaaaag ttgataacaa gtttaacaag gagcagcaga atgctttcta 1321 cgagatattg caccttccta atctgaatga ggaacaaaga aatgcattta tccagtccct 1381 aaaagacgat ccatcacaga gtgccaacct gcttgcggaa gccaaaaagc tcaatgatgc 1441 ccaggccccg aaagtagacg cgaaccatca ataagctagc ttgactgact gagatacagc 1501 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag 1561 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 1621 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 1681 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg 1741 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt 1801 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg 1861 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc 1921 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca 1981 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg 2041 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt 2101 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg 2161 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg 2221 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc 2281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 2341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 2401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 2461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 2521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 2581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 2641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 2701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 2761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 2821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 2881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 2941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 3001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 3061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 3121 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct 3181 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc 3241 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa 3301 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga 3361 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc 3421 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag 3481 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta 3541 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag 3601 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac 3661 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 3721 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca 3781 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc 3841 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 3901 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac 3961 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 4021 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg 4081 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 4141 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 4201 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt 4261 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 4321 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 4381 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 4441 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 4501 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg 4561 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact 4621 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga 4681 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt 4741 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg 4801 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac 4861 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag 4921 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga 4981 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta 5041 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 5101 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 5161 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 5221 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 5281 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 5341 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt 5401 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 5461 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 5521 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 5581 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca 5641 cccgccagcg gccgc //