LOCUS Exported 2634 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 2'-O-ribose methyltransferase. ACCESSION . VERSION . KEYWORDS pGBW-m4046823 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2634) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2634) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2634 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 20..39 /label=T7 /note="T7 promoter, forward primer" promoter 20..38 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 39..63 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 78..100 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 1005..1025 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1026..1055 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" rep_origin 1172..1760 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1661..1680 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1866..2525 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1932..1951) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2596..2630 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" ORIGIN 1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat 61 tcccctctag aaataatttt gtttaacttt aagaaggaga ctcttctatg tcaagccaag 121 catggcagcc cggggtggcc atgccaaact tgtacaaaat gcagcgtatg ttactggaga 181 agtgcgacct gcaaaattat ggtgattctg cgacgcttcc taaaggaatt atgatgaacg 241 tagctaaata cactcagctg tgtcagtatc tcaacaccct gacactggct gtcccgtaca 301 acatgcgcgt tatccacttc ggcgcaggct ccgataaagg tgttgcgccg ggcactgctg 361 tactgcgtca gtggctgccg accggtactc tgctggttga ctctgacctc aatgattttg 421 tgagtgacgc cgattccacc ctgatcggtg attgcgctac cgttcatact gcgaacaaat 481 gggacctgat tatcagcgat atgtacgacc cgaagaccaa aaacgtaacc aaagaaaacg 541 actcgaagga aggcttcttt acttatatct gtggtttcat tcagcagaaa ctggctcttg 601 gcggttcggt ggcaatcaag attacggaac actcttggaa cgctgaccta tacaaactta 661 tgggccattt cgcgtggtgg accgcctttg ttactaatgt caacgcgagc agctctgagg 721 cattcctgat cggatgcaat tacctgggta aaccgcgaga acagatcgat ggctatgtta 781 tgcacgctaa ctacatattc tggcgtaaca cgaacccgat ccaattatcc tcatattccc 841 tgtttgacat gagcaaattc ccactaaaac tgcgcggtac agcagtgatg tctctgaaag 901 aaggccagat taacgacatg attctgagtc tgctgtctaa agggagactg atcatccgtg 961 agaataaccg tgtggtcatc tcatccgatg ttctggtaaa caatgagaac ctgtactttc 1021 aaagcgagca gaagcttatt tccgaagaag atctctaacc ggcttatcgg tcagtttcac 1081 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa 1141 tgactgtcca cgacgctata cccaaaagaa attgagatcc tttttttctg cgcgtaatct 1201 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 1261 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 1321 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 1381 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 1441 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 1501 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 1561 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 1621 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 1681 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 1741 gggggcggag cctatggaaa ttgatcgggc acgtaagagg ttccaacttt caccataatg 1801 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag 1861 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta 1921 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc 1981 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct 2041 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag 2101 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa 2161 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca 2221 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta 2281 ttgagaatat gtttttcgtt tcagccaatc cctgggtgag tttcaccagt tttgatttaa 2341 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc 2401 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct 2461 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg 2521 cgtaatttga tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa 2581 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggt //