LOCUS Exported 2159 bp ds-DNA circular SYN 20-JAN-2023 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 nsp10. ACCESSION . VERSION . KEYWORDS pGBW-m4046840 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2159) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2159) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 20, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2159 /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 509..529 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 530..553 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" rep_origin 674..1262 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1163..1182 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1368..2027 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1434..1453) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2098..2132 /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 ggccggtaac gcaacggaag ttcccgcgaa 121 tagcacagtg ctgtcttttt gcgctttcgc tgtagacgcg gcaaaagctt acaaggatta 181 tttggcctca ggcggacagc caattaccaa ctgtgtcaaa atgctctgca ctcacaccgg 241 gactggccaa gctataaccg taactcctga ggcaaacatg gaccaggaat ccttcggtgg 301 tgcgagttgc tgcctgtact gtcgctgcca tatcgatcac ccgaacccga aaggcttttg 361 tgacctgaag gggaaatacg ttcagatccc gactacgtgc gctaacgatc cggttggttt 421 caccctgaaa aacaccgtgt gcaccgtgtg cggtatgtgg aaaggctatg gctgttcttg 481 tgaccagctg agggaaccga tgcttcaaga gaacctgtac tttcaaagct ggagccatcc 541 gcagtttgaa aaataaccgg cttatcggtc agtttcacct gatttacgta aaaacccgct 601 tcggcgggtt tttgcttttg gaggggcaga aagatgaatg actgtccacg acgctatacc 661 caaaagaaaa agcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 721 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 781 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 841 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 901 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 961 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 1021 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 1081 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 1141 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 1201 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 1261 aattgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg 1321 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa 1381 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat 1441 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt 1501 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc 1561 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat 1621 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc 1681 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg 1741 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg 1801 tttcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca 1861 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga 1921 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc 1981 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc 2041 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc 2101 agaacgctcg gttgccgccg ggcgtttttt attggtcatc gatctcgatc ccgcgaaat //