LOCUS Exported 1977 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4046786 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1977) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 1977) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1977 /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 354..374 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 375..398 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" rep_origin 515..1103 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1004..1023 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1209..1868 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1275..1294) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 1939..1973 /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 aaaatgtcag 121 atgtaaagtg cacatcagta gtcttactct cagttttgca acaactcaga gtagaatcat 181 catctaaatt gtgggctcaa tgtgtccagt tacacaatga cattctctta gctaaagata 241 ctactgaagc ctttgaaaaa atggtttcac tactttctgt tttgctttcc atgcagggtg 301 ctgtagacat aaacaagctt tgtgaagaaa tgctggacaa cagggcaacc ttagagaacc 361 tgtactttca aagcgactac aaggatgatg atgacaaata accggcttat cggtcagttt 421 cacctgattt acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg gcagaaagat 481 gaatgactgt ccacgacgct atacccaaaa gaaattgaga tccttttttt ctgcgcgtaa 541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 601 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 661 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 721 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 781 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 841 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 901 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 961 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 1021 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 1081 caggggggcg gagcctatgg aaattgatcg ggcacgtaag aggttccaac tttcaccata 1141 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg 1201 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc 1261 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc 1321 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg 1381 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga 1441 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc 1501 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac 1561 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt 1621 ttattgagaa tatgtttttc gtttcagcca atccctgggt gagtttcacc agttttgatt 1681 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata 1741 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg 1801 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg 1861 gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga tagatccagt aatgacctca 1921 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggt //