LOCUS Exported 7583 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 nsp3. ACCESSION . VERSION . KEYWORDS pGBW-m4046876 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7583) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7583) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7583 /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 5946..5966 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 5967..5996 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" rep_origin 6117..6705 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6606..6625 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 6811..7470 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(6877..6896) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 7541..7575 /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 gcccctacca 121 aggtgacgtt tggggatgac acagtcatcg aagttcaggg ttacaaaagc gtaaacatta 181 ctttcgagct ggacgaacgt atcgataaag ttcttaatga aaaatgctcc gcgtataccg 241 tggaattagg aactgaggtg aacgaatttg catgtgtagt tgctgacgct gttataaaaa 301 ctctacaacc ggtttcagaa ctcctgaccc cattgggcat cgatctggat gagtggtcta 361 tggcaaccta ctatctgttc gacgaaagtg gcgagttcaa gttggcgagc cacatgtact 421 gctctttcta cccgccggac gaagacgaag aagagggtga ttgtgaagag gaagaatttg 481 aaccctcgac tcagtatgaa tacggtacgg aagatgacta ccagggcaaa cctctggaat 541 ttggcgctac ctctgcggct ctgcaaccgg aagaggagca ggaagaggac tggctggatg 601 acgacagcca gcagactgtc ggtcagcagg atggttccga agataaccaa accacgacta 661 ttcagacaat cgtcgaagta caaccgcagc tggagatgga actgacccca gttgtacaga 721 ccatcgaagt gaactcgttc tctggctatc tgaaactgac ggacaatgtt tacattaaaa 781 acgcagatat cgtggaagaa gccaagaaag tgaaaccgac ggttgtagtt aacgctgcta 841 acgtatacct aaaacatggc ggcggtgtgg cgggtgcatt aaataaagcg actaacaatg 901 caatgcaggt tgaatccgat gactatatcg ctactaacgg tccgctgaag gtcggcggta 961 gctgcgttct gtctgggcac aacctggcaa aacattgcct gcacgtggta gggccgaacg 1021 tcaacaaagg cgaggatatt cagctgttaa agtcggccta cgaaaatttc aaccagcacg 1081 aagtactgct ggctccgctg ttaagtgctg gtatcttcgg tgccgaccct attcactccc 1141 tgagggtttg cgttgatacc gttcgcacca acgtctacct tgcagttttt gataagaacc 1201 tgtatgataa actggtgagc tctttcctgg aaatgaaatc tgaaaaacag gtagagcaga 1261 aaatcgccga aatcccaaaa gaggaagtaa agccgtttat tactgaaagc aagccgagcg 1321 ttgaacaacg taaacaggac gacaaaaaga tcaaagcttg tgtggaagaa gtcaccacaa 1381 ctcttgaaga aaccaaattc ctgactgaaa atctgctcct gtacatagac attaacggta 1441 acctgcatcc tgattccgcg actctggtga gtgacatcga tatcaccttc ctgaaaaagg 1501 acgcgcccta catcgttggc gacgtcgtac aagagggcgt tctcactgcg gtggttattc 1561 ccacgaaaaa agcaggcggt accacggaga tgttagccaa agctctgcgt aaagtgccta 1621 ccgacaacta tatcaccact tatccgggac aagggctgaa tggctacacc gtggaagaag 1681 ctaaaacggt actcaaaaaa tgcaaatccg ccttctacat cctgccgtcc atcatttcta 1741 acgagaaaca ggagatcctg ggtactgtca gctggaactt gcgcgaaatg ttggcacatg 1801 cggaggagac tcgtaaactc atgccggttt gcgttgaaac aaaggctatt gtttcaacca 1861 tccagcggaa atacaagggt attaagatcc aggaaggcgt cgtagattat ggagctcgct 1921 tctatttcta cactagcaaa acgacagtag cttctctgat taacaccctg aatgatctga 1981 acgaaaccct ggttactatg ccgctgggtt acgtaactca cggtctgaac ctggaagaag 2041 cagcccgata tatgcgttcc ctgaaggtac cggcaaccgt ttctgtatcg agtccagatg 2101 ccgtaacggc gtacaacggc tacctcacat ccagctctaa aaccccggaa gaacacttta 2161 tcgaaactat atcactggcg ggctcttaca aagattggag ttattccggt caaagcaccc 2221 agctgggaat tgaatttctg aagcgtggcg ataaatctgt ttactatact tcaaacccga 2281 ccactttcca cctggacggt gaagtaatca ctttcgacaa cttgaaaact ctgctgagct 2341 tgcgcgaggt ccgcaccatc aaagttttca ctactgtgga taacatcaac ctgcataccc 2401 aggtggttga catgtctatg acatacggcc agcagtttgg gccaacttac ctggacggcg 2461 cggacgttac caaaattaaa ccgcacaact cccatgaagg gaaaaccttc tacgtactgc 2521 ctaacgatga caccctgcgt gtggaagctt ttgaatacta ccacacgacc gacccgagtt 2581 tccttggtcg ttatatgagc gctttgaacc acaccaaaaa atggaagtac ccacaagtga 2641 acggtctgac tagcatcaaa tgggctgata acaattgcta cctagctacc gcactgctga 2701 cactgcaaca gatcgagctg aagttcaacc cgccggcgtt acaggatgct tactaccgtg 2761 ctcgcgccgg cgaagcagct aacttttgcg cactgattct ggcttattgt aacaaaaccg 2821 tgggcgaact gggtgatgtt cgtgaaacca tgagttacct tttccagcac gctaatctgg 2881 actcatgcaa acgcgtcctg aacgtggtat gtaaaacctg cggtcagcag caaaccacac 2941 tcaaaggcgt tgaagccgtt atgtatatgg gcactctgag ttatgagcag tttaaaaaag 3001 gtgtgcagat cccgtgtact tgcggcaaac aggctaccaa gtatctggtt cagcaggaat 3061 ctccattcgt aatgatgagc gcaccaccgg cgcagtacga actcaaacac gggactttta 3121 cttgcgcgtc tgaatacaca ggtaattacc agtgtggtca ctacaaacat attacgtcca 3181 aagaaactct ctattgcatc gacggcgctc tgctgaccaa atcgagcgaa tacaaaggcc 3241 ctatcacgga tgtcttctac aaagagaact cgtacacgac cactatcaaa ccggttacgt 3301 ataagctgga cggtgtagtc tgcactgaga tcgacccgaa attagataac tactacaaaa 3361 aagataactc ttattttaca gaacaaccaa ttgatctggt tccgaaccaa ccgtatccaa 3421 acgcgtcgtt cgataatttc aagttcgtct gcgacaacat aaaattcgcg gacgacttaa 3481 accagttaac agggtacaag aaacctgcgt cgcgtgaatt gaaagtcaca ttctttccgg 3541 atctgaatgg tgacgttgtt gcgatcgatt ataaacacta tactccgtcc ttcaaaaaag 3601 gagcaaaact gctgcataag ccgatcgttt ggcacgtaaa caacgctacc aataaagcaa 3661 cttataaacc gaacacttgg tgtattagat gcctgtggtc cactaaaccg gtggaaacct 3721 ctaatagctt tgatgtcctg aaatccgaag acgctcaggg tatggacaac ctagcttgcg 3781 aagacctgaa gccggtaagc gaggaggtcg ttgaaaatcc caccatccag aaagacgtac 3841 tggaatgcaa tgttaagact actgaagttg tcggagatat cattctgaaa cctgctaaca 3901 actctttgaa aatcacagag gaggtgggac acaccgatct gatggcagct tatgttgata 3961 acagctctct gaccatcaaa aagccgaatg aactaagccg tgtcctgggc ctgaaaactt 4021 tagccactca cggtttagcc gctgtgaact ctgttccctg ggataccatt gcaaactacg 4081 cgaaaccatt cctgaacaag gttgtgtcga ccactactaa tatcgtcacc cgctgcctga 4141 accgcgtgtg tactaactat atgccgtatt tcttcacgtt attactacaa ctgtgtacgt 4201 ttaccagatc gactaactcc cgaattaaag cttccatgcc gactaccatc gcgaaaaaca 4261 ccgttaaatc tgtcggcaaa ttctgcctgg aggcttcctt caactacctg aaatccccta 4321 acttttctaa gctcattaac ataatcatct ggttcctcct tctgtcggta tgcctaggtt 4381 cactgatcta cagtactgcg gccttgggcg ttctgatgag caatctgggc atgccgagct 4441 actgcactgg ttatcgcgag ggctacctta actctacgaa tgtgaccatc gcaacttact 4501 gtacgggttc aattccctgc tctgtttgtc tgagcggcct tgatagtctg gatacctatc 4561 cttcactgga gacaatccag attaccatct ctagctttaa atgggacctg accgcttttg 4621 gtctggttgc tgaatggttt ctggcgtaca tcctgtttac gcgtttcttt tatgttcttg 4681 gcctcgccgc cataatgcag ctgttcttct cctacttcgc agtccatttt atttcgaact 4741 cttggctgat gtggcttatc attaacctgg tgcagatggc tccgattagc gcaatggtac 4801 gtatgtacat ctttttcgct tctttctact acgtttggaa gtcttacgtg cacgtagtcg 4861 acggttgcaa tagctctacc tgtatgatgt gttataagcg taaccgtgct actcgcgtag 4921 aatgcactac tattgttaac ggtgttcgcc gcagcttcta tgtctacgcg aacggcggaa 4981 agggcttctg caagctgcac aactggaact gcgtgaattg cgatacattc tgtgctggtt 5041 ccacgtttat ctccgacgaa gtagcacggg atctgtctct gcaattcaaa cgtccgatca 5101 acccgaccga ccagtctagt tacatcgtgg attcagtcac tgttaaaaac ggctccattc 5161 atttatactt cgacaaggcg ggccagaaaa cctatgaacg tcattctctg agccacttcg 5221 tgaacttgga taatctgcgt gctaacaaca cgaaaggatc tctgccgatt aacgtaatcg 5281 ttttcgatgg taaaagtaaa tgcgaagaaa gtagcgcgaa atctgcttct gtgtactact 5341 cccaactgat gtgccagccg atcttactgc tggaccaggc tctggtttct gacgtgggtg 5401 acagcgccga agttgcggtt aaaatgttcg acgcatacgt aaacaccttc tcctctactt 5461 tcaacgtgcc aatggagaaa ctgaaaaccc tagtggcgac tgccgaggcg gaattggcta 5521 aaaacgtaag cctggacaat gttctgagca cctttatctc agcggcacgt caggggttcg 5581 tcgattctga cgtggagact aaagatgtag tcgaatgcct gaaacttagc catcagtccg 5641 acattgaagt tactggcgat tcgtgcaaca actatatgct gacctacaac aaagtggaga 5701 atatgacgcc gcgggacctg ggtgcttgca tcgactgctc tgcaaggcac atcaacgccc 5761 aggttgcaaa gtcgcataac atagctctga tctggaatgt taaggacttt atgtctctgt 5821 ctgaacagct gaggaagcaa attcgtagcg cggctaagaa aaataacctt ccgtttaaac 5881 tgacctgcgc aactactcgc caggtagtca atgtcgtgac cacgaagatc gcgctgaaag 5941 gtggtgagaa cctgtacttt caaagcgagc agaagcttat ttccgaagaa gatctctaac 6001 cggcttatcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg gtttttgctt 6061 ttggaggggc agaaagatga atgactgtcc acgacgctat acccaaaaga aaaagcttga 6121 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 6181 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 6241 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 6301 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 6361 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 6421 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 6481 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 6541 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 6601 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 6661 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaattgat cgggcacgta 6721 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta 6781 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc 6841 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa 6901 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa 6961 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat 7021 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct 7081 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac 7141 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac 7201 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtttcagc caatccctgg 7261 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt 7321 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag 7381 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag 7441 tactgcgatg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt 7501 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg 7561 ccgggcgttt tttattggtc atc //