LOCUS Exported 1958 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 E (envelope). ACCESSION . VERSION . KEYWORDS pGBW-m4046768 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1958) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 1958) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1958 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 75..109 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" primer_bind 137..156 /label=T7 /note="T7 promoter, forward primer" promoter 137..155 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 156..180 /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 195..217 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 450..470 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 471..488 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" rep_origin 609..1197 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1098..1117 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1303..4 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1369..1388) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" ORIGIN 1 gtaatttgat atcgagctcg cttggactcc tgttgataga tccagtaatg acctcagaac 61 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtcatcgat 121 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc 181 cctctagaaa taattttgtt taactttaag aaggagactc ttctatgtac tcattcgttt 241 cggaagagac aggtacgtta atagttaata gcgtacttct ttttcttgct ttcgtggtat 301 tcttgctagt tacactagcc atccttactg cgcttcgatt gtgtgcgtac tgctgcaata 361 ttgttaacgt gagtcttgta aaaccttctt tttacgttta ctctcgtgtt aaaaatctga 421 attcttctag agttcctgat cttctggtcg agaacctgta ctttcaaagc caccatcatc 481 accatcatta accggcttat cggtcagttt cacctgattt acgtaaaaac ccgcttcggc 541 gggtttttgc ttttggaggg gcagaaagat gaatgactgt ccacgacgct atacccaaaa 601 gaaaaagctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 1021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 1081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 1141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaattg 1201 atcgggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta 1261 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact 1321 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag 1381 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag 1441 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg 1501 atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat 1561 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg 1621 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt 1681 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtttca 1741 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc 1801 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg 1861 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat 1921 gaattacaac agtactgcga tgagtggcag ggcggggc //