LOCUS Exported 7577 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 nsp3. ACCESSION . VERSION . KEYWORDS pGBW-m4046947 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7577) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7577) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7577 /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..5990 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" rep_origin 6111..6699 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6600..6619 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 6805..7464 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(6871..6890) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 7535..7569 /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 caaagctgga gccatccgca gtttgaaaaa taaccggctt 6001 atcggtcagt ttcacctgat ttacgtaaaa acccgcttcg gcgggttttt gcttttggag 6061 gggcagaaag atgaatgact gtccacgacg ctatacccaa aagaaaaagc ttgagatcct 6121 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 6181 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 6241 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 6301 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 6361 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 6421 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 6481 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 6541 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 6601 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 6661 tttttgtgat gctcgtcagg ggggcggagc ctatggaaat tgatcgggca cgtaagaggt 6721 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga 6781 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat 6841 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc 6901 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag 6961 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa 7021 tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac 7081 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat 7141 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc 7201 tatttcccta aagggtttat tgagaatatg tttttcgttt cagccaatcc ctgggtgagt 7261 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc 7321 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat 7381 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc 7441 gatgagtggc agggcggggc gtaatttgat atcgagctcg cttggactcc tgttgataga 7501 tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc 7561 gttttttatt ggtcatc //