LOCUS Exported 1991 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4046832 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1991) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 1991) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1991 /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 360..380 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 381..404 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" rep_origin 525..1113 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1014..1033 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1219..1878 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1285..1304) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 1949..1983 /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 agcaaaatgt 121 ctgatgtgaa gtgtacttcc gttgtattgc tgtcagtcct gcaacagctg cgtgttgagt 181 cgtctagcaa actatgggca cagtgcgtgc agctgcacaa cgacatcctc ttagcgaaag 241 atacgaccga agctttcgaa aaaatggttt cgctgttgtc cgtacttctg tctatgcaag 301 gtgccgttga cattaataaa ctgtgcgaag agatgctgga caaccgcgct actctgcaag 361 agaacctgta ctttcaaagc tggagccatc cgcagtttga aaaataaccg gcttatcggt 421 cagtttcacc tgatttacgt aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag 481 aaagatgaat gactgtccac gacgctatac ccaaaagaaa aagcttgaga tccttttttt 541 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 601 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 661 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 721 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 781 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 841 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 901 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 961 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 1021 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 1081 tgatgctcgt caggggggcg gagcctatgg aaattgatcg ggcacgtaag aggttccaac 1141 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca 1201 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc 1261 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac 1321 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag 1381 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt 1441 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt 1501 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg 1561 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc 1621 cctaaagggt ttattgagaa tatgtttttc gtttcagcca atccctgggt gagtttcacc 1681 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc 1741 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc 1801 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag 1861 tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga tagatccagt 1921 aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt 1981 tattggtcat c //