LOCUS Exported 2070 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression plasmid for SARS-CoV-2 nsp3. ACCESSION . VERSION . KEYWORDS pGBW-m4095419 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2070) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2070) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2070 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 434..454 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 455..472 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" rep_origin 589..1177 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1078..1097 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1283..1942 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1349..1368) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2013..2047 /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag actcttctat gttcacagag caaccaattg atcttgtacc 121 aaaccaacca tatccaaacg caagcttcga taattttaag tttgtatgtg ataatatcaa 181 atttgctgat gatttaaacc agttaactgg ttataagaaa cctgcttcaa gagagcttaa 241 agttacgttt ttccctgact taaatggtga tgtggtggct attgattata aacactacac 301 accctctttt aagaaaggag ctaaattgtt acataaacct attgtttggc atgttaacaa 361 tgcaactaat aaagccacgt ataaaccaaa tacctggtgt atacgttgtc tttggagcac 421 aaaaccagtt gaagagaacc tgtactttca aagccaccat catcaccatc attaaccggc 481 ttatcggtca gtttcacctg atttacgtaa aaacccgctt cggcgggttt ttgcttttgg 541 aggggcagaa agatgaatga ctgtccacga cgctataccc aaaagaaatt gagatccttt 601 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 661 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 721 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 781 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 841 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 901 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 961 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 1021 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 1081 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 1141 tttgtgatgc tcgtcagggg ggcggagcct atggaaattg atcgggcacg taagaggttc 1201 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt 1261 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat 1321 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta 1381 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca 1441 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt 1501 tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac 1561 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt 1621 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta 1681 tttccctaaa gggtttattg agaatatgtt tttcgtttca gccaatccct gggtgagttt 1741 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat 1801 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca 1861 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga 1921 tgagtggcag ggcggggcgt aatttgatat cgagctcgct tggactcctg ttgatagatc 1981 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt 2041 tttttattgg tgatctcgat cccgcgaaat //