LOCUS Exported 3309 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 3'-to-5' exonuclease. ACCESSION . VERSION . KEYWORDS pGBW-m4046902 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3309) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 3309) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3309 /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 1686..1706 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1707..1730 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" rep_origin 1847..2435 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2336..2355 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2541..3200 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(2607..2626) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 3271..3305 /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 gaaaatgtaa 121 caggactctt taaagattgt agtaaggtaa tcactgggtt acatcctaca caggcaccta 181 cacacctcag tgttgacact aaattcaaaa ctgaaggttt atgtgttgac atacctggca 241 tacctaagga catgacctat agaagactca tctctatgat gggttttaaa atgaattatc 301 aagttaatgg ttaccctaac atgtttatca cccgcgaaga agctataaga catgtacgtg 361 catggattgg cttcgatgtc gaggggtgtc atgctactag agaagctgtt ggtaccaatt 421 tacctttaca gctaggtttt tctacaggtg ttaacctagt tgctgtacct acaggttatg 481 ttgatacacc taataataca gatttttcca gagttagtgc taaaccaccg cctggagatc 541 aatttaaaca cctcatacca cttatgtaca aaggacttcc ttggaatgta gtgcgtataa 601 agattgtaca aatgttaagt gacacactta aaaatctctc tgacagagtc gtatttgtct 661 tatgggcaca tggctttgag ttgacatcta tgaagtattt tgtgaaaata ggacctgagc 721 gcacctgttg tctatgtgat agacgtgcca catgcttttc cactgcttca gacacttatg 781 cctgttggca tcattctatt ggatttgatt acgtctataa tccgtttatg attgatgttc 841 aacaatgggg ttttacaggt aacctacaaa gcaaccatga tctgtattgt caagtccatg 901 gtaatgcaca tgtagctagt tgtgatgcaa tcatgactag gtgtctagct gtccacgagt 961 gctttgttaa gcgtgttgac tggactattg aatatcctat aattggtgat gaactgaaga 1021 ttaatgcggc ttgtagaaag gttcaacaca tggttgttaa agctgcatta ttagcagaca 1081 aattcccagt tcttcacgac attggtaacc ctaaagctat taagtgtgta cctcaagctg 1141 atgtagaatg gaagttctat gatgcacagc cttgtagtga caaagcttat aaaatagaag 1201 aattattcta ttcttatgcc acacattctg acaaattcac agatggtgta tgcctatttt 1261 ggaattgcaa tgtcgataga tatcctgcta attccattgt ttgtagattt gacactagag 1321 tgctatctaa ccttaacttg cctggttgtg atggtggcag tttgtatgta aataaacatg 1381 cattccacac accagctttt gataaaagtg cttttgttaa tttaaaacaa ttaccatttt 1441 tctattactc tgacagtcca tgtgagtctc atggaaaaca agtagtgtca gatatagatt 1501 atgtaccact aaagtctgct acgtgtataa cacgttgcaa tttaggtggt gctgtctgta 1561 gacatcatgc taatgagtac agattgtatc tcgatgctta taacatgatg atctcagctg 1621 gctttagctt gtgggtttac aaacaatttg atacttataa cctctggaac acttttacaa 1681 gacttgagaa cctgtacttt caaagcgact acaaggatga tgatgacaaa taaccggctt 1741 atcggtcagt ttcacctgat ttacgtaaaa acccgcttcg gcgggttttt gcttttggag 1801 gggcagaaag atgaatgact gtccacgacg ctatacccaa aagaaattga gatccttttt 1861 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 1921 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 1981 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 2041 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 2101 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 2161 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 2221 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 2281 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 2341 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 2401 tgtgatgctc gtcagggggg cggagcctat ggaaattgat cgggcacgta agaggttcca 2461 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt 2521 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat 2581 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata 2641 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca 2701 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc 2761 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg 2821 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc 2881 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt 2941 tccctaaagg gtttattgag aatatgtttt tcgtttcagc caatccctgg gtgagtttca 3001 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg 3061 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg 3121 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg 3181 agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt gatagatcca 3241 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt 3301 tttattggt //