LOCUS Exported 2336 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 nsp8. ACCESSION . VERSION . KEYWORDS pGBW-m4046793 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2336) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2336) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2336 /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 699..719 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 720..749 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" rep_origin 870..1458 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1359..1378 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1564..2223 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1630..1649) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2294..2328 /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 atagcctcag 121 agtttagttc ccttccatca tatgcagctt ttgctactgc tcaagaagct tatgagcagg 181 ctgttgctaa tggtgattct gaagttgttc ttaaaaagtt gaagaagtct ttgaatgtgg 241 ctaaatctga atttgaccgt gatgcagcca tgcaacgtaa gttggaaaag atggctgatc 301 aagctatgac ccaaatgtat aaacaggcta gatctgagga caagagggca aaagttacta 361 gtgctatgca gacaatgctt ttcactatgc ttagaaagtt ggataatgat gcactcaaca 421 acattatcaa caatgcaaga gatggttgtg ttcccttgaa cataatacct cttacaacag 481 cagccaaact aatggttgtc ataccagact ataacacata taaaaatacg tgtgatggta 541 caacatttac ttatgcatca gcattgtggg aaatccaaca ggttgtagat gcagatagta 601 aaattgttca acttagtgaa attagtatgg acaattcacc taatttagca tggcctctta 661 ttgtaacagc tttaagggcc aattctgctg tcaaattaga gaacctgtac tttcaaagcg 721 agcagaagct tatttccgaa gaagatctct aaccggctta tcggtcagtt tcacctgatt 781 tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg ggcagaaaga tgaatgactg 841 tccacgacgc tatacccaaa agaaaaagct tgagatcctt tttttctgcg cgtaatctgc 901 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 961 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 1021 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 1081 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 1141 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 1201 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 1261 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 1321 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 1381 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 1441 gggcggagcc tatggaaatt gatcgggcac gtaagaggtt ccaactttca ccataatgaa 1501 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct 1561 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa 1621 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg 1681 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt 1741 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac 1801 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact 1861 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata 1921 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt 1981 gagaatatgt ttttcgtttc agccaatccc tgggtgagtt tcaccagttt tgatttaaac 2041 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa 2101 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc 2161 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg 2221 taatttgata tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact 2281 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtcatc //