LOCUS Exported 6855 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4252964 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6855) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6855) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6855 /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..319 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" polyA_signal 388..509 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(399..418) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 453..472 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 624..670 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 700..836 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 881..952 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1178..1766) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1258..1277) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2266..3475 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3312..3331 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 3479..4507 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA KE" polyA_signal 4534..4582 /note="synthetic polyadenylation signal" CDS 4848..5708 /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(5066..5085) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 5721..5857 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 6009..6080 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 6085..6131 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 6149..6197 /note="synthetic polyadenylation signal" enhancer 6278..6581 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6583..6785 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6736..6756 /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 tcagtcgggg aaacctatac ccaatccact 301 tctcggctta gattctacct aaggatccgc aggcctctgc tagcttgact gactgagata 361 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa 421 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg 481 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc 541 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta 601 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag 661 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg 721 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg 781 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc 841 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt 901 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt 961 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga 1021 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct 1081 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa 1141 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 1201 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 1261 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 1321 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 1381 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 1441 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 1501 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 1561 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa 1621 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 1681 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 1741 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 1801 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 1861 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 1921 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 1981 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa 2041 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag 2101 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact 2161 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc 2221 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg 2281 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag 2341 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata 2401 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga 2461 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct 2521 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc 2581 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt 2641 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg 2701 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg 2761 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct 2821 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg 2881 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag 2941 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac 3001 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg 3061 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct 3121 gttggcttat aatgcagggt ggggccacca gccggtaggt gtgcggtagg cttttctccg 3181 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct 3241 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct 3301 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg 3361 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg 3421 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat 3481 gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga 3541 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga 3601 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga 3661 tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg attccggaag tgcttgacat 3721 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt 3781 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga 3841 tgccatcgct gccgccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg 3901 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta 3961 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga 4021 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg 4081 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc 4141 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc 4201 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc 4261 ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga 4321 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg 4381 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct ggaccgatgg 4441 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa 4501 ggaataagct agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg 4561 atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc 4621 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag 4681 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta 4741 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag 4801 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac 4861 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 4921 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca 4981 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc 5041 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 5101 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac 5161 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 5221 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg 5281 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 5341 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 5401 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt 5461 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 5521 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 5581 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 5641 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 5701 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg 5761 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact 5821 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga 5881 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt 5941 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg 6001 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac 6061 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag 6121 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga 6181 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta 6241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 6301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 6361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 6421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 6481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 6541 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt 6601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 6661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 6721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 6781 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca 6841 cccgccagcg gccgc //