LOCUS Exported 3551 bp ds-DNA circular SYN 20-JAN-2023 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 helicase. ACCESSION . VERSION . KEYWORDS pGBW-m4046911 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3551) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 3551) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 20, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3551 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 1895..1915 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1916..1945 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" rep_origin 2066..2654 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2555..2574 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2760..3419 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(2826..2845) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 3490..3524 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag actcttctat ggctgtgggt gcatgtgtct tatgcaactc 121 acagaccagc cttcgctgcg gcgcgtgcat tcgtcgtccc ttcctgtgtt gcaaatgctg 181 ttatgaccat gtaatcagta cgtcgcacaa gctggttctg tccgttaatc cgtacgtttg 241 caacgctccg ggatgtgatg taacagacgt gactcaactc tacctgggtg gcatgtctta 301 ctattgcaaa agccacaaac caccgatatc ctttcctctg tgcgccaacg gccaggtatt 361 cggtctgtac aaaaatacct gtgttgggtc tgataacgtg actgacttta acgcgatcgc 421 aacttgcgat tggaccaacg ctggcgacta tatcctggcg aatacctgca ctgagcgatt 481 gaaactgttc gctgctgaaa ctctgaaggc gaccgaagaa accttcaaac tctcctacgg 541 tattgcaacg gtccgtgagg ttctatctga tcgtgaactg catctgagct gggaagtagg 601 taagccgcgc ccgcctctga accgcaacta cgtttttact ggctatcgtg ttacaaaaaa 661 ctctaaagtg cagatcggtg aatacacctt cgagaaaggg gactacggcg atgcagttgt 721 ctatcggggc accactacat ataagttaaa cgtcggtgac tacttcgttc tgacttcaca 781 cacggtgatg ccactgtccg ctccgactct ggttccgcag gaacactacg tgcgcattac 841 tgggctgtac ccgacattga acatctcgga tgagttctcc tctaacgtag ccaattacca 901 gaaagtaggt atgcaaaaat acagcactct gcaaggcccg cctggtaccg gaaaaagcca 961 tttcgctata ggtctggccc tgtattaccc atccgcgcgc atcgtctaca ctgcatgcag 1021 tcacgctgcg gtggacgcac tgtgcgaaaa ggcccttaaa tatctcccta ttgataaatg 1081 ttctcgtatc atcccggcga gagctcgtgt tgaatgcttt gacaaattta aagttaacag 1141 caccctggag cagtacgtgt tctgtaccgt taatgcactg ccggaaacta ccgctgatat 1201 tgtggtgttc gacgaaatct ctatggcgac caactacgat ctgagcgtag ttaacgcgcg 1261 tttacgtgct aaacactacg tatacatcgg cgacccagca cagctgccgg caccgaggac 1321 tctgcttact aaaggcaccc tggaaccgga gtacttcaac tcagtttgcc gtctgatgaa 1381 gacgatcggt ccagatatgt ttcttggcac gtgtcgcaga tgcccggcag aaattgtgga 1441 taccgtatct gctctggttt atgataacaa actgaaagcc cataaggaca agtccgctca 1501 gtgcttcaaa atgttttata aaggtgttat cacgcacgac gtcagtagcg cgatcaaccg 1561 tccacagatt ggagttgtgc gagaatttct cacccgcaat ccggcatggc gtaaagctgt 1621 cttcatctct ccctataact ctcagaacgc agtagcctcc aaaatcctgg gtctgccgac 1681 gcagactgtg gattctagcc agggctcaga atatgattac gtgattttca cccaaaccac 1741 tgaaaccgct cactcctgca acgttaaccg gttcaacgta gctattaccc gcgccaaagt 1801 tggtatcctc tgcatcatgt ctgaccgcga cctgtacgac aaactgcaat ttacgagcct 1861 cgaaatcccc cgccgtaacg tagctacttt acaggagaac ctgtactttc aaagcgagca 1921 gaagcttatt tccgaagaag atctctaacc ggcttatcgg tcagtttcac ctgatttacg 1981 taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa tgactgtcca 2041 cgacgctata cccaaaagaa aaagcttgag atcctttttt tctgcgcgta atctgctgct 2101 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 2161 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 2221 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 2281 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 2341 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 2401 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 2461 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 2521 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 2581 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 2641 ggagcctatg gaaattgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa 2701 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa 2761 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac 2821 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata 2881 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc 2941 acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg 3001 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa 3061 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt 3121 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga 3181 atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg 3241 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg 3301 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg 3361 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat 3421 ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat 3481 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtca tcgatctcga 3541 tcccgcgaaa t //