LOCUS Exported 4538 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 RNA-dependent RNA polymerase. ACCESSION . VERSION . KEYWORDS pGBW-m4046831 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4538) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4538) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4538 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 5..24 /label=T7 /note="T7 promoter, forward primer" promoter 5..23 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 24..48 /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 63..85 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 2886..2906 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 2907..2936 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" rep_origin 3057..3645 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3546..3565 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3751..4410 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(3817..3836) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 4481..4515 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" ORIGIN 1 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata 61 attttgttta actttaagaa ggagactctt ctatggctga tgcacaatcg tttttaaacc 121 gggtttgcgg tgtaagtgca gcccgtctta caccgtgcgg cacaggcact agtactgatg 181 tcgtatacag ggcttttgac atctacaatg ataaagtagc tggttttgct aaattcctaa 241 aaactaattg ttgtcgcttc caagaaaagg acgaagatga caatttaatt gattcttact 301 ttgtagttaa gagacacact ttctctaact accaacatga agaaacaatt tataatttac 361 ttaaggattg tccagctgtt gctaaacatg acttctttaa gtttagaata gacggtgaca 421 tggtaccaca tatatcacgt caacgtctta ctaaatacac aatggcagac ctcgtctatg 481 ctttaaggca ttttgatgaa ggtaattgtg acacattaaa agaaatactt gtcacataca 541 attgttgtga tgatgattat ttcaataaaa aggactggta tgattttgta gaaaacccag 601 atatattacg cgtatacgcc aacttaggtg aacgtgtacg ccaagctttg ttaaaaacag 661 tacaattctg tgatgccatg cgaaatgctg gtattgttgg tgtactgaca ttagataatc 721 aagatctcaa tggtaactgg tatgatttcg gtgatttcat acaaaccacg ccaggtagtg 781 gagttcctgt tgtagattct tattattcat tgttaatgcc tatattaacc ttgaccaggg 841 ctttaactgc agagtcacat gttgacactg acttaacaaa gccttacatt aagtgggatt 901 tgttaaaata tgacttcacg gaagagaggt taaaactctt tgaccgttat tttaaatatt 961 gggatcagac ataccaccca aattgtgtta actgtttgga tgacagatgc attctgcatt 1021 gtgcaaactt taatgtttta ttctctacag tgttcccacc tacaagtttt ggaccactag 1081 tgagaaaaat atttgttgat ggtgttccat ttgtagtttc aactggatac cacttcagag 1141 agctaggtgt tgtacataat caggatgtaa acttacatag ctctagactt agttttaagg 1201 aattacttgt gtatgctgct gaccctgcta tgcacgctgc ttctggtaat ctattactag 1261 ataaacgcac tacgtgcttt tcagtagctg cacttactaa caatgttgct tttcaaactg 1321 tcaaacccgg taattttaac aaagacttct atgactttgc tgtgtctaag ggtttcttta 1381 aggaaggaag ttctgttgaa ttaaaacact tcttctttgc tcaggatggt aatgctgcta 1441 tcagcgatta tgactactat cgttataatc taccaacaat gtgtgatatc agacaactac 1501 tatttgtagt tgaagttgtt gataagtact ttgattgtta cgatggtggc tgtattaatg 1561 ctaaccaagt catcgtcaac aacctagaca aatcagctgg ttttccattt aataaatggg 1621 gtaaggctag actttattat gattcaatga gttatgagga tcaagatgca cttttcgcat 1681 atacaaaacg taatgtcatc cctactataa ctcaaatgaa tcttaagtat gccattagtg 1741 caaagaatag agctcgcacc gtagctggtg tctctatctg tagtactatg accaatagac 1801 agtttcatca aaaattattg aaatcaatag ccgccactag aggagctact gtagtaattg 1861 gaacaagcaa attctatggt ggttggcaca acatgttaaa aactgtttat agtgatgtag 1921 aaaaccctca ccttatgggt tgggattatc ctaaatgtga tagagccatg cctaacatgc 1981 ttagaattat ggcctcactt gttcttgctc gcaaacatac aacgtgttgt agcttgtcac 2041 accgtttcta tagattagct aatgagtgtg ctcaagtatt gagtgaaatg gtcatgtgtg 2101 gcggttcact atatgttaaa ccaggtggaa cctcatcagg agatgccaca actgcttatg 2161 ctaatagtgt ttttaacatt tgtcaagctg tcacggccaa tgttaatgca cttttatcta 2221 ctgatggtaa caaaattgcc gataagtatg tccgcaattt acaacacaga ctttatgagt 2281 gtctctatag aaatagagat gttgacacag actttgtgaa tgagttttac gcatatttgc 2341 gtaaacattt ctcaatgatg atactctctg acgatgctgt tgtgtgtttc aatagcactt 2401 atgcatctca aggtctagtg gctagcataa agaactttaa gtcagttctt tattatcaaa 2461 acaatgtttt tatgtctgaa gcaaaatgtt ggactgagac tgaccttact aaaggacctc 2521 atgaattttg ctctcaacat acaatgctag ttaaacaggg tgatgattat gtgtaccttc 2581 cttacccaga tccatcaaga atcctagggg ccggctgttt tgtagatgat atcgtaaaaa 2641 cagatggtac acttatgatt gaacggttcg tgtctttagc tatagatgct tacccactta 2701 ctaaacatcc taatcaggag tatgctgatg tctttcattt gtacttacaa tacataagaa 2761 agctacatga tgagttaaca ggacacatgt tagacatgta ttctgttatg cttactaatg 2821 ataacacttc aaggtattgg gaacctgagt tttatgaggc tatgtacaca ccgcatacag 2881 tcttagagaa cctgtacttt caaagcgagc agaagcttat ttccgaagaa gatctctaac 2941 cggcttatcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg gtttttgctt 3001 ttggaggggc agaaagatga atgactgtcc acgacgctat acccaaaaga aaaagcttga 3061 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 3121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 3181 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 3241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 3301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 3361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 3421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 3481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 3541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 3601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaattgat cgggcacgta 3661 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta 3721 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc 3781 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa 3841 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa 3901 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat 3961 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct 4021 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac 4081 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac 4141 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtttcagc caatccctgg 4201 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt 4261 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag 4321 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag 4381 tactgcgatg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt 4441 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg 4501 ccgggcgttt tttattggtc atcgatctcg atcccgcg //