LOCUS Exported 3539 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 helicase. ACCESSION . VERSION . KEYWORDS pGBW-m4046760 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" RBS 1786..1808 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind 1823..1847 /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)." primer_bind complement(1847..1866) /label=T7 /note="T7 promoter, forward primer" promoter complement(1848..1866) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" terminator 1894..1928 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" CDS complement(1999..2658) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 2573..2592 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" rep_origin complement(2764..3352) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2844..2863) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3473..3490) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS complement(3491..3511) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" ORIGIN 1 atttcgaggc tcgtaaattg cagtttgtcg tacaggtcgc ggtcagacat gatgcagagg 61 ataccaactt tggcgcgggt aatagctacg ttgaaccggt taacgttgca ggagtgagcg 121 gtttcagtgg tttgggtgaa aatcacgtaa tcatattctg agccctggct agaatccaca 181 gtctgcgtcg gcagacccag gattttggag gctactgcgt tctgagagtt atagggagag 241 atgaagacag ctttacgcca tgccggattg cgggtgagaa attctcgcac aactccaatc 301 tgtggacggt tgatcgcgct actgacgtcg tgcgtgataa cacctttata aaacattttg 361 aagcactgag cggacttgtc cttatgggct ttcagtttgt tatcataaac cagagcagat 421 acggtatcca caatttctgc cgggcatctg cgacacgtgc caagaaacat atctggaccg 481 atcgtcttca tcagacggca aactgagttg aagtactccg gttccagggt gcctttagta 541 agcagagtcc tcggtgccgg cagctgtgct gggtcgccga tgtatacgta gtgtttagca 601 cgtaaacgcg cgttaactac gctcagatcg tagttggtcg ccatagagat ttcgtcgaac 661 accacaatat cagcggtagt ttccggcagt gcattaacgg tacagaacac gtactgctcc 721 agggtgctgt taactttaaa tttgtcaaag cattcaacac gagctctcgc cgggatgata 781 cgagaacatt tatcaatagg gagatattta agggcctttt cgcacagtgc gtccaccgca 841 gcgtgactgc atgcagtgta gacgatgcgc gcggatgggt aatacagggc cagacctata 901 gcgaaatggc tttttccggt accaggcggg ccttgcagag tgctgtattt ttgcatacct 961 actttctggt aattggctac gttagaggag aactcatccg agatgttcaa tgtcgggtac 1021 agcccagtaa tgcgcacgta gtgttcctgc ggaaccagag tcggagcgga cagtggcatc 1081 accgtgtgtg aagtcagaac gaagtagtca ccgacgttta acttatatgt agtggtgccc 1141 cgatagacaa ctgcatcgcc gtagtcccct ttctcgaagg tgtattcacc gatctgcact 1201 ttagagtttt ttgtaacacg atagccagta aaaacgtagt tgcggttcag aggcgggcgc 1261 ggcttaccta cttcccagct cagatgcagt tcacgatcag atagaacctc acggaccgtt 1321 gcaataccgt aggagagttt gaaggtttct tcggtcgcct tcagagtttc agcagcgaac 1381 agtttcaatc gctcagtgca ggtattcgcc aggatatagt cgccagcgtt ggtccaatcg 1441 caagttgcga tcgcgttaaa gtcagtcacg ttatcagacc caacacaggt atttttgtac 1501 agaccgaata cctggccgtt ggcgcacaga ggaaaggata tcggtggttt gtggcttttg 1561 caatagtaag acatgccacc caggtagagt tgagtcacgt ctgttacatc acatcccgga 1621 gcgttgcaaa cgtacggatt aacggacaga accagcttgt gcgacgtact gattacatgg 1681 tcataacagc atttgcaaca caggaaggga cgacgaatgc acgcgccgca gcgaaggctg 1741 gtctgtgagt tgcataagac acatgcaccc acagccatag aagagtctcc ttcttaaagt 1801 taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc 1861 gtattaattt cgcgggatcg agatcgatga ccaataaaaa acgcccggcg gcaaccgagc 1921 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca 1981 agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc 2041 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag 2101 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa 2161 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc 2221 tgaaacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta 2281 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact 2341 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact 2401 atcccatatc accagctcac cgtctttcat tgccatacga aattccggat gagcattcat 2461 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt 2521 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga 2581 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc 2641 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa 2701 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat 2761 caatttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 2821 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 2881 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 2941 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 3001 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 3061 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 3121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 3181 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 3241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 3301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagctttttc 3361 ttttgggtat agcgtcgtgg acagtcattc atctttctgc ccctccaaaa gcaaaaaccc 3421 gccgaagcgg gtttttacgt aaatcaggtg aaactgaccg ataagccggt taatgatggt 3481 gatgatggtg gctttgaaag tacaggttct cctgtaaagt agctacgtta cggcggggg //