LOCUS Exported 6406 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4133943 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6406) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6406) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6406 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 257..277 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 278..322 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" polyA_signal 375..496 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(386..405) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 440..459 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 611..657 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 687..823 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 868..939 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1165..1753) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1245..1264) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2253..3462 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3299..3318 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 3466..4065 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3466..3485) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3962..3982 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 4092..4140 /note="synthetic polyadenylation signal" CDS 4406..5266 /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(4624..4643) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 5279..5415 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 5560..5631 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 5636..5682 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 5700..5748 /note="synthetic polyadenylation signal" enhancer 5829..6132 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6134..6336 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6287..6307 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagc aaaatgtctg acgtcaagtg tacatccgta gttctgttat cagtgcttca 61 acagctgcgg gtggagagtt cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga 121 tatattgctc gcgaaagata ctacggaagc tttcgagaag atggtcagct tgctatctgt 181 tctactgagc atgcaaggag cagtagacat caacaaactt tgcgaagaga tgttagacaa 241 ccgagccacc ctgcaagaaa acctttactt tcagtcgaaa gaaaccgcgg cagctaagtt 301 cgagaggcag cacatggaca gttaagctag cttgactgac tgagatacag cgtaccttca 361 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg 421 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag 481 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga 541 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct 601 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc 661 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc 721 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg 781 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg 841 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta 901 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc 961 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt 1021 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc 1081 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg 1141 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 1201 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 1261 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 1321 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 1381 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 1441 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 1501 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 1561 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 1621 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 1681 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 1741 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 1801 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 1861 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 1921 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 1981 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt 2041 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc 2101 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata 2161 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga 2221 gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc 2281 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt 2341 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta 2401 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg 2461 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg 2521 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca 2581 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat 2641 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc 2701 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa 2761 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt 2821 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt 2881 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt 2941 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact 3001 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt 3061 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat 3121 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag 3181 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag 3241 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg 3301 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag 3361 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa 3421 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag 3481 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 3541 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 3601 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 3661 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 3721 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 3781 cagcaacaga tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 3841 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 3901 cccggagtgg aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 3961 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 4021 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc 4081 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg 4141 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat 4201 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat 4261 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata 4321 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga 4381 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc 4441 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 4501 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg 4561 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt 4621 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 4681 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac 4741 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 4801 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa 4861 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc 4921 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt 4981 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga 5041 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa 5101 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa 5161 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa 5221 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc 5281 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca 5341 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca 5401 gaaatttatc cttaagatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc 5461 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta 5521 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca 5581 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa 5641 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac 5701 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg 5761 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg 5821 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 5881 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 5941 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 6001 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 6061 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 6121 gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 6181 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 6241 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 6301 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga 6361 tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgc //