LOCUS Exported 2646 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 3C-like proteinase. ACCESSION . VERSION . KEYWORDS pGBW-m4046922 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2646) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2646) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2646 /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 1023..1043 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1044..1067 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" rep_origin 1184..1772 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1673..1692 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1878..2537 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1944..1963) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2608..2642 /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 ggttttagaa 121 aaatggcatt cccatctggt aaagttgagg gttgtatggt acaagtaact tgtggtacaa 181 ctacacttaa cggtctttgg cttgatgacg tagtttactg tccaagacat gtgatctgca 241 cctctgaaga catgcttaac cctaattatg aagatttact cattcgtaag tctaatcata 301 atttcttggt acaggctggt aatgttcaac tcagggttat tggacattct atgcaaaatt 361 gtgtacttaa gcttaaggtt gatacagcca atcctaagac acctaagtat aagtttgttc 421 gcattcaacc aggacagact ttttcagtgt tagcttgtta caatggttca ccatctggtg 481 tttaccaatg tgctatgagg cccaatttca ctattaaggg ttcattcctt aatggttcat 541 gtggtagtgt tggttttaac atagattatg actgtgtctc tttttgttac atgcaccata 601 tggaattacc aactggagtt catgctggca cagacttaga aggtaacttt tatggacctt 661 ttgttgacag gcaaacagca caagcagctg gtacggacac aactattaca gttaatgttt 721 tagcttggtt gtacgctgct gttataaatg gagacaggtg gtttctcaat cgatttacca 781 caactcttaa tgactttaac cttgtggcta tgaagtacaa ttatgaacct ctaacacaag 841 accatgttga catactagga cctctttctg ctcaaactgg aattgccgtt ttagatatgt 901 gtgcttcatt aaaagaatta ctgcaaaatg gtatgaatgg acgtaccata ttgggtagtg 961 ctttattaga agatgaattt acaccttttg atgttgttag acaatgctca ggtgttactt 1021 tcgagaacct gtactttcaa agcgactaca aggatgatga tgacaaataa ccggcttatc 1081 ggtcagtttc acctgattta cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg 1141 cagaaagatg aatgactgtc cacgacgcta tacccaaaag aaattgagat cctttttttc 1201 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 1261 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 1321 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 1381 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 1441 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 1501 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 1561 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 1621 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 1681 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 1741 gatgctcgtc aggggggcgg agcctatgga aattgatcgg gcacgtaaga ggttccaact 1801 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag 1861 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc 1921 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc 1981 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt 2041 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta 2101 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt 2161 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc 2221 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc 2281 ctaaagggtt tattgagaat atgtttttcg tttcagccaa tccctgggtg agtttcacca 2341 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca 2401 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg 2461 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt 2521 ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac tcctgttgat agatccagta 2581 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt 2641 attggt //