LOCUS Exported 2274 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 leader protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046821 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2274) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2274) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2274 /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 645..665 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 666..695 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" rep_origin 812..1400 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1301..1320 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1506..2165 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1572..1591) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2236..2270 /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 gagagccttg 121 tccctggttt caacgagaaa acacacgtcc aactcagttt gcctgtttta caggttcgcg 181 acgtgctcgt acgtggcttt ggagactccg tggaggaggt cttatcagag gcacgtcaac 241 atcttaaaga tggcacttgt ggcttagtag aagttgaaaa aggcgttttg cctcaacttg 301 aacagcccta tgtgttcatc aaacgttcgg atgctcgaac tgcacctcat ggtcatgtta 361 tggttgagct ggtagcagaa ctcgaaggca ttcagtacgg tcgtagtggt gagacacttg 421 gtgtccttgt ccctcatgtg ggcgaaatac cagtggctta ccgcaaggtt cttcttcgta 481 agaacggtaa taaaggagct ggtggccata gttacggcgc cgatctaaag tcatttgact 541 taggcgacga gcttggcact gatccttatg aagattttca agaaaactgg aacactaaac 601 atagcagtgg tgttacccgt gaactcatgc gtgagcttaa cggagagaac ctgtactttc 661 aaagcgagca gaagcttatt tccgaagaag atctctaacc ggcttatcgg tcagtttcac 721 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa 781 tgactgtcca cgacgctata cccaaaagaa attgagatcc tttttttctg cgcgtaatct 841 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 901 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 961 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 1021 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 1081 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 1141 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 1201 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 1261 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 1321 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 1381 gggggcggag cctatggaaa ttgatcgggc acgtaagagg ttccaacttt caccataatg 1441 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag 1501 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta 1561 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc 1621 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct 1681 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag 1741 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa 1801 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca 1861 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta 1921 ttgagaatat gtttttcgtt tcagccaatc cctgggtgag tttcaccagt tttgatttaa 1981 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc 2041 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct 2101 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg 2161 cgtaatttga tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa 2221 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggt //