LOCUS Exported 2654 bp ds-DNA circular SYN 01-APR-2022 DEFINITION Bacterial expression plasmid for SARS-CoV-2 3C-like proteinase. ACCESSION . VERSION . KEYWORDS pGBW-m4046747 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2654) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2654) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 1, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2654 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 1010..1030 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1031..1048 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" rep_origin 1169..1757 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1658..1677 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1863..2522 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1929..1948) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2593..2627 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag actcttctat gagcggcttt cgcaaaatgg ccttcccttc 121 cggtaaggtg gaggggtgca tggtccaggt aacttgtggc acgaccactc tgaacggttt 181 atggcttgac gatgttgttt attgcccacg gcacgttatc tgtacaagtg aagatatgct 241 gaatcccaac tacgaagacc tgctcattcg taaatcgaac cataacttct tggtgcaagc 301 aggcaatgtg cagctgcgtg tcatcggtca ctctatgcag aactgcgtac taaaactgaa 361 agttgacacc gcgaacccga aaaccccgaa gtacaaattc gtacgcatcc aaccgggaca 421 gacttttagc gttctggctt gctataacgg ctctccgagc ggtgtttacc agtgcgctat 481 gcgaccgaac ttcactatta agggttcttt tctgaatggc tcctgtggaa gtgtaggctt 541 caacatagat tacgactgcg tcagcttttg ttatatgcac catatggaac tgccaaccgg 601 tgttcacgcg ggtacagatc tggagggcaa cttctacggt ccgttcgttg accgccagac 661 cgctcaagca gcggggacgg acaccactat cacggtgaat gtgctggcct ggctgtacgc 721 agctgtgatc aacggcgatc gttggtttct gaacagattc accactactc ttaacgattt 781 caacctcgtt gctatgaaat acaactacga accgctgacc caggatcacg ttgacatttt 841 aggtcctctg tcagctcaga ccgggatcgc agtactggat atgtgcgcgt ctttaaaaga 901 acttctgcaa aatggcatga atggtcgtac tattctgggc tccgccctgc tggaggacga 961 atttactccg ttcgacgtag tacgtcagtg ctctggtgta acttttcagg agaacctgta 1021 ctttcaaagc caccatcatc accatcatta accggcttat cggtcagttt cacctgattt 1081 acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg gcagaaagat gaatgactgt 1141 ccacgacgct atacccaaaa gaaaaagctt gagatccttt ttttctgcgc gtaatctgct 1201 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 1261 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 1321 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 1381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 1441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 1501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 1561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 1621 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 1681 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 1741 ggcggagcct atggaaattg atcgggcacg taagaggttc caactttcac cataatgaaa 1801 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta 1861 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag 1921 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg 1981 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta 2041 ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg 2101 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg 2161 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat 2221 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg 2281 agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt gatttaaacg 2341 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag 2401 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc 2461 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt 2521 aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc 2581 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tcatcgatct 2641 cgatcccgcg aaat //