LOCUS Exported 1779 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4046725 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1779) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 1779) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1779 /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 150..170 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 171..200 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" rep_origin 317..905 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 806..825 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1011..1670 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1077..1096) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 1741..1775 /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 agcgctgatg 121 cgcaaagttt tctcaatggt ttcgccgtag agaacctgta ctttcaaagc gagcagaagc 181 ttatttccga agaagatctc taaccggctt atcggtcagt ttcacctgat ttacgtaaaa 241 acccgcttcg gcgggttttt gcttttggag gggcagaaag atgaatgact gtccacgacg 301 ctatacccaa aagaaattga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 361 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 421 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 481 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 541 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 601 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 661 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 721 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 781 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 841 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 901 ggaaattgat cgggcacgta agaggttcca actttcacca taatgaaata agatcactac 961 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa 1021 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg 1081 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct 1141 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg 1201 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga 1261 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat 1321 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg 1381 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt 1441 tcgtttcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg 1501 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc 1561 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa 1621 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttgatatcg 1681 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg 1741 ttcagaacgc tcggttgccg ccgggcgttt tttattggt //