LOCUS Exported 2640 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 3C-like proteinase. ACCESSION . VERSION . KEYWORDS pGBW-m4046754 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2640) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2640) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2640 /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..1061 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" rep_origin 1178..1766 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1667..1686 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1872..2531 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1938..1957) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2602..2636 /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 agccaccatc atcaccatca ttaaccggct tatcggtcag 1081 tttcacctga tttacgtaaa aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa 1141 gatgaatgac tgtccacgac gctataccca aaagaaattg agatcctttt tttctgcgcg 1201 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 1261 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 1321 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 1381 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 1441 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 1501 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 1561 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 1621 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 1681 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 1741 cgtcaggggg gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc 1801 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta 1861 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc 1921 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg 1981 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc 2041 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa 2101 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg 2161 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc 2221 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag 2281 ggtttattga gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg 2341 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt 2401 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg 2461 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg 2521 gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc 2581 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt //