LOCUS Exported 2598 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 nsp6. ACCESSION . VERSION . KEYWORDS pGBW-m4046916 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2598) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2598) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2598 /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 981..1001 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1002..1019 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" rep_origin 1136..1724 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1625..1644 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1830..2489 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1896..1915) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2560..2594 /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 tccgcggtga 121 agcgtacgat taaaggcact caccattggc tgttacttac aatcctaacc agcctgctgg 181 tcctcgttca aagtactcag tggtcgctgt tctttttctt gtatgagaac gccttcctgc 241 cgtttgcaat ggggatcata gctatgtcag cgttcgctat gatgtttgtt aaacacaaac 301 atgcattctt atgtctgttt ctgctgccaa gcctggctac cgtagcatac ttcaatatgg 361 tttacatgcc ggcttcctgg gtgatgcgca ttatgacttg gctggatatg gtagacacct 421 ctctgtctgg tttcaagctt aaagactgcg ttatgtatgc cagcgcggtg gttctgctga 481 tcctgatgac cgctaggacc gtctacgatg atggtgctcg tcgcgtatgg actctaatga 541 acgttctgac tcttgtatac aaagtctatt acggcaacgc actggaccag gcgatctcta 601 tgtgggccct gatcatttct gttacatcca attacagtgg tgtagttacc acggtaatgt 661 tccttgcccg tggcattgtg ttcatgtgcg tggaatattg ccctatcttc tttatcactg 721 gtaacaccct gcaatgtatc atgttggttt actgctttct gggctacttt tgcacttgct 781 atttcgggct gttctgttta ctcaaccgtt actttcgcct gaccctgggt gtctacgact 841 acctggttag cacacaggaa tttcgctaca tgaacagcca gggactgctt cccccgaaaa 901 attctattga tgctttcaaa ctgaacatca agttgttggg cgtgggtggc aaaccgtgca 961 tcaaagtggc gacggttcag gagaacctgt actttcaaag ccaccatcat caccatcatt 1021 aaccggctta tcggtcagtt tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg 1081 cttttggagg ggcagaaaga tgaatgactg tccacgacgc tatacccaaa agaaattgag 1141 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 1201 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 1261 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 1321 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 1381 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 1441 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 1501 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 1561 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 1621 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 1681 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaattgatc gggcacgtaa 1741 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat 1801 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg 1861 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat 1921 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa 1981 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc 2041 cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt 2101 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg 2161 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc 2221 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtttcagcc aatccctggg 2281 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt 2341 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg 2401 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt 2461 actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg actcctgttg 2521 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc 2581 cgggcgtttt ttattggt //