LOCUS Exported 2606 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 nsp6. ACCESSION . VERSION . KEYWORDS pGBW-m4046966 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2606) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2606) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2606 /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 975..995 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 996..1019 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" rep_origin 1140..1728 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1629..1648 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1834..2493 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1900..1919) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2564..2598 /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 gcagtgaaaa 121 gaacaatcaa gggtacacac cactggttgt tactcacaat tttgacttca cttttagttt 181 tagtccagag tactcaatgg tctttgttct tttttttgta tgaaaatgcc tttttacctt 241 ttgctatggg tattattgct atgtctgctt ttgcaatgat gtttgtcaaa cataagcatg 301 catttctctg tttgtttttg ttaccttctc ttgccactgt agcttatttt aatatggtct 361 atatgcctgc tagttgggtg atgcgtatta tgacatggtt ggatatggtt gatactagtt 421 tgtctggttt taagctaaaa gactgtgtta tgtatgcatc agctgtagtg ttactaatcc 481 ttatgacagc aagaactgtg tatgatgatg gtgctaggag agtgtggaca cttatgaatg 541 tcttgacact cgtttataaa gtttattatg gtaatgcttt agatcaagcc atttccatgt 601 gggctcttat aatctctgtt acttctaact actcaggtgt agttacaact gtcatgtttt 661 tggccagagg tattgttttt atgtgtgttg agtattgccc tattttcttc ataactggta 721 atacacttca gtgtataatg ctagtttatt gtttcttagg ctatttttgt acttgttact 781 ttggcctctt ttgtttactc aaccgctact ttagactgac tcttggtgtt tatgattact 841 tagtttctac acaggagttt agatatatga attcacaggg actactccca cccaagaata 901 gcatagatgc cttcaaactc aacattaaat tgttgggtgt tggtggcaaa ccttgtatca 961 aagtagccac tgtagagaac ctgtactttc aaagctggag ccatccgcag tttgaaaaat 1021 aaccggctta tcggtcagtt tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg 1081 cttttggagg ggcagaaaga tgaatgactg tccacgacgc tatacccaaa agaaaaagct 1141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 1201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 1261 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 1321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 1381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 1441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 1501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 1561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 1621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 1681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaatt gatcgggcac 1741 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag 1801 ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc 1861 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct 1921 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag 1981 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct 2041 catccggaat ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac 2101 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac 2161 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa 2221 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtttc agccaatccc 2281 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc 2341 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt 2401 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa 2461 cagtactgcg atgagtggca gggcggggcg taatttgata tcgagctcgc ttggactcct 2521 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg 2581 ccgccgggcg ttttttattg gtcatc //