LOCUS Exported 6849 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4252899 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6849) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6849) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6849 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 251..271 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 272..313 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" polyA_signal 382..503 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(393..412) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 447..466 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 618..664 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 694..830 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 875..946 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1172..1760) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1252..1271) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2260..3469 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3306..3325 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 3473..4501 /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 4528..4576 /note="synthetic polyadenylation signal" CDS 4842..5702 /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(5060..5079) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 5715..5851 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 6003..6074 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 6079..6125 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 6143..6191 /note="synthetic polyadenylation signal" enhancer 6272..6575 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6577..6779 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6730..6750 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca 61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat 121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt 181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag 241 ggcaacctta gaaaaccttt actttcagtc ggggaaacct atacccaatc cacttctcgg 301 cttagattct acctaaggat ccgcaggcct ctgctagctt gactgactga gatacagcgt 361 accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa 421 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca 481 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc 541 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc 601 catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt 661 tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat 721 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc 781 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc 841 tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac 901 accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac 961 cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc 1021 tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca 1081 cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca 1141 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 1201 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 1261 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 1321 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 1381 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 1441 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 1501 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 1561 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 1621 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 1681 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 1741 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 1801 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 1861 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 1921 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 1981 gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc 2041 aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt 2101 ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc 2161 ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc 2221 tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg 2281 cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca 2341 ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc 2401 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag 2461 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga 2521 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg 2581 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg 2641 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc 2701 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg 2761 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc 2821 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg 2881 gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta 2941 ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg 3001 gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg 3061 cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc 3121 ttataatgca gggtggggcc accagccggt aggtgtgcgg taggcttttc tccgtcgcag 3181 gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg 3241 aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa 3301 gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt 3361 tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac 3421 ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgggtaa 3481 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt 3541 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg 3601 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta 3661 tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg acattgggga 3721 gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga 3781 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa tggatgccat 3841 cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg 3901 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg 3961 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat 4021 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa 4081 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt 4141 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat 4201 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccccggct 4261 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa 4321 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg 4381 gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg atggctgtgt 4441 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata 4501 agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt 4561 gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag 4621 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg 4681 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc 4741 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg 4801 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc 4861 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 4921 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac 4981 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga 5041 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 5101 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca 5161 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 5221 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa 5281 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 5341 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 5401 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag 5461 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 5521 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 5581 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 5641 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 5701 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat 5761 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata 5821 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg 5881 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt 5941 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc 6001 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc 6061 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta 6121 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata 6181 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt 6241 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 6301 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 6361 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 6421 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 6481 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 6541 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca 6601 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca 6661 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg 6721 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg 6781 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc 6841 agcggccgc //