LOCUS Exported 2276 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for SARS-CoV-2 leader protein. ACCESSION . VERSION . KEYWORDS pGBW-m4046770 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2276) TITLE SARS-CoV-2 protein E. coli expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 2276) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2276 /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..689 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" rep_origin 810..1398 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1299..1318 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 1504..2163 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1570..1589) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2234..2268 /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 aaagctggag ccatccgcag tttgaaaaat aaccggctta tcggtcagtt tcacctgatt 721 tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg ggcagaaaga tgaatgactg 781 tccacgacgc tatacccaaa agaaaaagct tgagatcctt tttttctgcg cgtaatctgc 841 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 901 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 961 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 1021 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 1081 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 1141 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 1201 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 1261 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 1321 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 1381 gggcggagcc tatggaaatt gatcgggcac gtaagaggtt ccaactttca ccataatgaa 1441 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct 1501 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa 1561 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg 1621 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt 1681 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac 1741 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact 1801 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata 1861 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt 1921 gagaatatgt ttttcgtttc agccaatccc tgggtgagtt tcaccagttt tgatttaaac 1981 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa 2041 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc 2101 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg 2161 taatttgata tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact 2221 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtcatc //