LOCUS Exported 3542 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 helicase. ACCESSION . VERSION . KEYWORDS pGBW-m4046898 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3542) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 3542) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3542 /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 1908..1928 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1929..1955 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" rep_origin 2076..2664 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2565..2584 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2770..3429 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(2836..2855) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 3500..3534 /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 gttggggctt 121 gtgttctttg caattcacag acttcattaa gatgtggtgc ttgcatacgt agaccattct 181 tatgttgtaa atgctgttac gaccatgtca tatcaacatc acataaatta gtcttgtctg 241 ttaatccgta tgtttgcaat gctccaggtt gtgatgtcac agatgtgact caactttact 301 taggaggtat gagctattat tgtaaatcac ataaaccacc cattagtttt ccattgtgtg 361 ctaatggaca agtttttggt ttatataaaa atacatgtgt tggtagcgat aatgttactg 421 actttaatgc aattgcaaca tgtgactgga caaatgctgg tgattacatt ttagctaaca 481 cctgtactga aagactcaag ctttttgcag cagaaacgct caaagctact gaggagacat 541 ttaaactgtc ttatggtatt gctactgtac gtgaagtgct gtctgacaga gaattacatc 601 tttcatggga agttggtaaa cctagaccac cacttaaccg aaattatgtc tttactggtt 661 atcgtgtaac taaaaacagt aaagtacaaa taggagagta cacctttgaa aaaggtgact 721 atggtgatgc tgttgtttac cgaggtacaa caacttacaa attaaatgtt ggtgattatt 781 ttgtgctgac atcacataca gtaatgccat taagtgcacc tacactagtg ccacaagagc 841 actatgttag aattactggc ttatacccaa cactcaatat ctcagatgag ttttctagca 901 atgttgcaaa ttatcaaaag gttggtatgc aaaagtattc tacactccag ggaccacctg 961 gtactggtaa gagtcatttt gctattggcc tagctctcta ctacccttct gctcgcatag 1021 tgtatacagc ttgctctcat gccgctgttg atgcactatg tgagaaggca ttaaaatatt 1081 tgcctataga taaatgtagt agaattatac ctgcacgtgc tcgtgtagag tgttttgata 1141 aattcaaagt gaattcaaca ttagaacagt atgtcttttg tactgtaaat gcattgcctg 1201 agacgacagc agatatagtt gtctttgatg aaatttcaat ggccacaaat tatgatttga 1261 gtgttgtcaa tgccagatta cgtgctaagc actatgtgta cattggcgac cctgctcaat 1321 tacctgcacc acgcacattg ctaactaagg gcacactaga accagaatat ttcaattcag 1381 tgtgtagact tatgaaaact ataggtccag acatgttcct cggaacttgt cggcgttgtc 1441 ctgctgaaat tgttgacact gtgagtgctt tggtttatga taataagctt aaagcacata 1501 aagacaaatc agctcaatgc tttaaaatgt tttataaggg tgttatcacg catgatgttt 1561 catctgcaat taacaggcca caaataggcg tggtaagaga attccttaca cgtaaccctg 1621 cttggagaaa agctgtcttt atttcacctt ataattcaca gaatgctgta gcctcaaaga 1681 ttttgggact accaactcaa actgttgatt catcacaggg ctcagaatat gactatgtca 1741 tattcactca aaccactgaa acagctcact cttgtaatgt aaacagattt aatgttgcta 1801 ttaccagagc aaaagtaggc atactttgca taatgtctga tagagacctt tatgacaagt 1861 tgcaatttac aagtcttgaa attccacgta ggaatgtggc aactttagag aacctgtact 1921 ttcaaagcta cccgtatgac gttcctgatt atgcctaacc ggcttatcgg tcagtttcac 1981 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa 2041 tgactgtcca cgacgctata cccaaaagaa aaagcttgag atcctttttt tctgcgcgta 2101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 2161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 2221 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 2281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 2341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 2401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 2461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 2521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 2581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 2641 tcaggggggc ggagcctatg gaaattgatc gggcacgtaa gaggttccaa ctttcaccat 2701 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag 2761 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat 2821 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt 2881 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg 2941 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg 3001 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag 3061 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta 3121 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg 3181 tttattgaga atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat 3241 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat 3301 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat 3361 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc 3421 ggggcgtaat ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc 3481 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtca 3541 tc //