LOCUS Exported 3539 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 helicase. ACCESSION . VERSION . KEYWORDS pGBW-m4046813 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3539) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 3539) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3539 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 24..43 /label=T7 /note="T7 promoter, forward primer" promoter 24..42 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 43..67 /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 82..104 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 1912..1932 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1933..1956 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" rep_origin 2077..2665 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2566..2585 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2771..3430 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(2837..2856) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 3501..3535 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" ORIGIN 1 catcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa 61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagactcttc tatggttggg 121 gcttgtgttc tttgcaattc acagacttca ttaagatgtg gtgcttgcat acgtagacca 181 ttcttatgtt gtaaatgctg ttacgaccat gtcatatcaa catcacataa attagtcttg 241 tctgttaatc cgtatgtttg caatgctcca ggttgtgatg tcacagatgt gactcaactt 301 tacttaggag gtatgagcta ttattgtaaa tcacataaac cacccattag ttttccattg 361 tgtgctaatg gacaagtttt tggtttatat aaaaatacat gtgttggtag cgataatgtt 421 actgacttta atgcaattgc aacatgtgac tggacaaatg ctggtgatta cattttagct 481 aacacctgta ctgaaagact caagcttttt gcagcagaaa cgctcaaagc tactgaggag 541 acatttaaac tgtcttatgg tattgctact gtacgtgaag tgctgtctga cagagaatta 601 catctttcat gggaagttgg taaacctaga ccaccactta accgaaatta tgtctttact 661 ggttatcgtg taactaaaaa cagtaaagta caaataggag agtacacctt tgaaaaaggt 721 gactatggtg atgctgttgt ttaccgaggt acaacaactt acaaattaaa tgttggtgat 781 tattttgtgc tgacatcaca tacagtaatg ccattaagtg cacctacact agtgccacaa 841 gagcactatg ttagaattac tggcttatac ccaacactca atatctcaga tgagttttct 901 agcaatgttg caaattatca aaaggttggt atgcaaaagt attctacact ccagggacca 961 cctggtactg gtaagagtca ttttgctatt ggcctagctc tctactaccc ttctgctcgc 1021 atagtgtata cagcttgctc tcatgccgct gttgatgcac tatgtgagaa ggcattaaaa 1081 tatttgccta tagataaatg tagtagaatt atacctgcac gtgctcgtgt agagtgtttt 1141 gataaattca aagtgaattc aacattagaa cagtatgtct tttgtactgt aaatgcattg 1201 cctgagacga cagcagatat agttgtcttt gatgaaattt caatggccac aaattatgat 1261 ttgagtgttg tcaatgccag attacgtgct aagcactatg tgtacattgg cgaccctgct 1321 caattacctg caccacgcac attgctaact aagggcacac tagaaccaga atatttcaat 1381 tcagtgtgta gacttatgaa aactataggt ccagacatgt tcctcggaac ttgtcggcgt 1441 tgtcctgctg aaattgttga cactgtgagt gctttggttt atgataataa gcttaaagca 1501 cataaagaca aatcagctca atgctttaaa atgttttata agggtgttat cacgcatgat 1561 gtttcatctg caattaacag gccacaaata ggcgtggtaa gagaattcct tacacgtaac 1621 cctgcttgga gaaaagctgt ctttatttca ccttataatt cacagaatgc tgtagcctca 1681 aagattttgg gactaccaac tcaaactgtt gattcatcac agggctcaga atatgactat 1741 gtcatattca ctcaaaccac tgaaacagct cactcttgta atgtaaacag atttaatgtt 1801 gctattacca gagcaaaagt aggcatactt tgcataatgt ctgatagaga cctttatgac 1861 aagttgcaat ttacaagtct tgaaattcca cgtaggaatg tggcaacttt agagaacctg 1921 tactttcaaa gctggagcca tccgcagttt gaaaaataac cggcttatcg gtcagtttca 1981 cctgatttac gtaaaaaccc gcttcggcgg gtttttgctt ttggaggggc agaaagatga 2041 atgactgtcc acgacgctat acccaaaaga aaaagcttga gatccttttt ttctgcgcgt 2101 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 2161 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 2221 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 2281 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 2341 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 2401 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 2461 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 2521 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 2581 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 2641 gtcagggggg cggagcctat ggaaattgat cgggcacgta agaggttcca actttcacca 2701 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa 2761 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca 2821 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt 2881 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc 2941 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat 3001 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga 3061 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct 3121 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg 3181 gtttattgag aatatgtttt tcgtttcagc caatccctgg gtgagtttca ccagttttga 3241 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta 3301 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga 3361 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg 3421 cggggcgtaa tttgatatcg agctcgcttg gactcctgtt gatagatcca gtaatgacct 3481 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggt //