LOCUS Exported 5175 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 3C-like proteinase. ACCESSION . VERSION . KEYWORDS pGBW-m4133367 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5175) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5175) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5175 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 926..946 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 947..991 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" polyA_signal 1044..1165 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1055..1074) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1109..1128 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1280..1326 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1356..1492 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1537..1608 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1834..2422) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1914..1933) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3168..4028 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3386..3405) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4041..4177 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4329..4400 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4405..4451 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4469..4517 /note="synthetic polyadenylation signal" enhancer 4598..4901 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4903..5105 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5056..5076 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagt gggttccgga aaatggcgtt tccctccggt aaggtcgaag gatgcatggt 61 tcaggtgacc tgtggcacaa ctacgttaaa cggcctatgg ctggacgatg tggtatactg 121 tcctcgacac gtcatctgca catcagagga catgctgaat ccaaactatg aggatttgct 181 cataagaaag agcaatcata actttctggt gcaagctgga aatgtacagc ttagggttat 241 tgggcactct atgcagaact gcgtgttaaa actcaaggtg gataccgcca atccgaaaac 301 acctaagtac aaattcgtgc gcatccagcc aggccaaacc ttctctgtcc tggcatgtta 361 taacggcagc ccctcaggtg tctaccagtg cgccatgcgc cctaatttta ctattaaggg 421 ttccttcttg aacgggtcgt gcggatcggt cggctttaat atcgactatg actgtgtgag 481 tttttgctac atgcatcaca tggaacttcc gactggggtt cacgctggaa cggatctgga 541 gggtaacttc tatggaccct tcgtggatag acagaccgcc caggcagccg gcactgacac 601 taccattacg gtgaatgttc tagcttggct ctacgcagct gtaataaacg gcgatcgttg 661 gttcttgaac aggtttacaa caaccctgaa cgactttaat ctagtcgcca tgaaatataa 721 ttatgaacca ctgacccaag atcatgtaga cattctcggg ccactgagcg cacagacagg 781 gatcgctgtg ctggacatgt gtgccagttt aaaggagctg cttcagaatg gtatgaacgg 841 ccgaacaatc ctagggtctg cgctcttaga ggatgaattt acccccttcg atgtagtgcg 901 gcaatgtagc ggcgttactt ttcaagaaaa cctttacttt cagtcgaaag aaaccgcggc 961 agctaagttc gagaggcagc acatggacag ttaagctagc ttgactgact gagatacagc 1021 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag 1081 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 1141 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 1201 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg 1261 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt 1321 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg 1381 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc 1441 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca 1501 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg 1561 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt 1621 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg 1681 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg 1741 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc 1801 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 1861 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 1921 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 1981 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 2041 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 2101 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 2161 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 2221 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 2281 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 2341 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 2401 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 2461 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 2521 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 2581 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 2641 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct 2701 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc 2761 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa 2821 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga 2881 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc 2941 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag 3001 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta 3061 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag 3121 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac 3181 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 3241 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca 3301 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc 3361 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 3421 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac 3481 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 3541 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg 3601 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 3661 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 3721 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt 3781 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 3841 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 3901 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 3961 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 4021 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg 4081 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact 4141 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga 4201 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt 4261 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg 4321 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac 4381 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag 4441 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga 4501 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta 4561 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 4621 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 4681 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 4741 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 4801 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 4861 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt 4921 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 4981 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 5041 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 5101 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca 5161 cccgccagcg gccgc //